Method for producing oligonucleic acid compound

Abstract

The present invention relates to a method for producing an oligonucleic acid compound, the method being characterized by (1) treating a compound represented by formula [A-1] and a compound represented by formula [B-1] with a condensing agent in the presence of a base and then (2) treating the compounds with an oxidizing agent and an organic amine to produce a compound represented by formula [C-1] (in the formulae, each symbol is as defined in the description).

Description

[Technical Field]

[0001] The present invention relates to a method for producing a novel oligonucleic acid compound. [Background technology]

[0002] Known methods for producing oligonucleic acid compounds include solid-phase and liquid-phase methods. The solid-phase method is a heterogeneous reaction method in which nucleic acids are extended while contacting a substrate supported on a solid support with a solution containing a reaction reagent. The solid-phase method uses a reaction vessel equipped with a filter, a so-called batch method in which the reaction is carried out in the vessel (see, for example, Non-Patent Document 1 and Patent Document 1). A pseudo-flow synthesis method, similar to that used in automated nucleic acid synthesizers (e.g., DNA and RNA synthesizers), is also known in which the solid support is placed in a column and a solution containing a reaction reagent is passed through the column to cause the reaction. On the other hand, the liquid phase method is a homogeneous reaction method in which nucleic acids are elongated by reacting them in a solution containing both a substrate and a reaction reagent. The liquid phase method also uses a batch method in which the reaction is carried out in a container (see, for example, Patent Document 2 and Patent Document 3).

[0003] Whether solid-phase, liquid-phase, batch, or pseudo-flow synthesis is used, chemical synthesis of oligonucleic acid compounds involves repeatedly repeating a "deprotection" reaction, which removes the protecting groups on the oxygen atoms or amino groups on the nucleic acid compound, and a "condensation" reaction, which forms a bond between the oxygen atom or nitrogen atom that has been deprotected and is now reactive, and the phosphorus atom, thereby extending the nucleic acid. In particular, controlling the reaction efficiency and reaction rate in the "condensation" reaction, which forms a bond between a phosphorus atom and an oxygen atom or a nitrogen atom, is extremely important in the production of oligonucleic acid compounds, and the conditions for this condensation reaction are factors that have a significant impact on the production period of oligonucleic acid compounds.

[0004] Because the solid-phase method is a heterogeneous reaction between a solid support and a solution, it is known that the reactivity of the condensation reaction decreases due to steric hindrance caused by the solid support. Polystyrene resin is generally used as the solid support, but during the reaction, it swells in the reaction solvent used, and its volume increases compared to its dry state. The degree of swelling depends on the reaction solvent. Therefore, the reaction efficiency and reaction rate of the condensation reaction in the solid phase method depend on the reaction solvent used. In particular, the swelling degree of polystyrene resin is not so large in polar solvents such as acetonitrile, which is generally used in the synthesis of oligonucleic acid compounds, and therefore the use of polar solvents in the solid phase method is not preferable from the viewpoint of improving the reaction efficiency and reaction rate of the condensation reaction.

[0005] On the other hand, as a homogeneous reaction method, a liquid phase method and a synthesis method using a hydrophobic group-bound nucleoside or a pseudo-solid phase protected nucleoside are known. The liquid-phase method is a homogeneous reaction method in which the reaction is carried out in a solution containing both the substrate and the reaction reagent. Compared to the solid-phase method, the reaction efficiency is higher and the reaction rate is faster, but column purification is required to remove impurities such as the reaction reagent and reaction solvent. Similar to the liquid-phase method, synthesis methods using hydrophobic-group-bound nucleosides or pseudo-solid-phase protected nucleosides can be carried out in a homogeneous system, resulting in higher reaction efficiency and faster reaction rates than solid-phase methods. Furthermore, after the reaction, unnecessary reaction reagents and reaction solvents can be removed by precipitating the target compound from the reaction mixture (see, for example, Patent Document 4). [Prior art documents] [Patent documents]

[0006] [Patent Document 1] International Publication No. 1991 / 09033A1 [Patent Document 2] International Publication No. 2014 / 077292A1 [Patent Document 3] International Publication No. 2013 / 122236A1 [Patent Document 4] Japanese Patent No. 5548852 [Patent Document 5] International Publication No. 2016 / 060135A1 [Non-patent literature]

[0007] [Non-Patent Document 1] Acc.Chem.Res.,Vol.24,278-284,1991 Summary of the Invention [Problem to be solved by the invention]

[0008] An object of the present invention is to provide a novel production method that can shorten the production period of an oligonucleic acid compound. [Means for solving the problem]

[0009] The present inventors have discovered that the formation of an H-phosphonate in the condensation reaction of an oligonucleic acid compound allows the condensation reaction to proceed efficiently, and have thus completed the present invention.

[0010] That is, the present invention relates to the following:

[0011] <1> Formula [A-1]: [ka] [In the formula, B P are the same or different, optionally protected nucleobases; n is an integer of 1 to 50, preferably an integer of 1 to 30, more preferably an integer of 1 to 25, and most preferably an integer of 1 to 15; W are the same or different and each is an oxygen atom or a sulfur atom; and X's may be the same or different and each represent a hydroxyl group substituted with a group that can be removed under neutral conditions, 1,1,3,3-tetra(C 1-6 Alkyl)guanidyl, C 1-6Alkoxy, di(C 1-6 Alkyl)amino, mono(amino-C substituted with a group that can be removed under basic conditions 1-6 Alkyl)amino, di(amino-C substituted with a group that can be removed under basic conditions 1-6 alkyl)amino, or general formula [2]: [ka] (In the formula, * is the position of attachment to P; a is an integer from 0 to 2; E is CH2, CH-A 1 or NA 2 and; A 1 is C 1-6 Alkyl, mono(C 1-6 Alkyl)amino-C 1-6 Alkyl, di(C 1-6 Alkyl)amino-C 1-6 Alkyl, Tri(C 1-6 Alkyl)ammonio-C 1-6 Alkyl, amino substituted with a group removable under basic conditions, mono(C 1-6 alkyl)amino, di(C 1-6 alkyl)amino, tri(C 1-6 alkyl)ammonio, amino substituted with amidino substituted with a group removable under basic conditions, or general formula [3]: [ka] (In the formula, * is the bonding position with E; b is an integer from 0 to 2; c is 0 or 1; R 11 is C 1-6 is alkyl; and M is a substituent represented by CH, an oxygen atom, a sulfur atom, or N- (a group removable under basic conditions); and A 2is C 1-6 Alkyl, mono(C 1-6 Alkyl)amino-C 1-6 Alkyl, di(C 1-6 Alkyl)amino-C 1-6 Alkyl, Tri(C 1-6 Alkyl)ammonio-C 1-6 alkyl, a group removable under basic conditions, aryl, or heteroaryl) is a substituent represented by the formula: G is (1) silicon substituents, (2) optionally substituted C 1-18 Alkyl-carbonyl, optionally substituted C 1-18 alkoxy-carbonyl, long chain alkyl-carbonyl, or long chain alkoxy-carbonyl; (3) Benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; or (4) General formula [7]: [ka] (In the formula, * is the binding position to T; Z is (1) (soluble polymer soluble in organic solvent)-oxy, (2) (Soluble polymer soluble in organic solvent)-amino, (3) long-chain alkyloxy, benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or benzyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; (4) a solid support, or (5) The following general formulas [8A] to [8N]: [ka] (In the formula, * indicates the position of attachment to L; j is an integer from 0 to 4; k is an integer from 0 to 5; R 8a is a hydrogen atom or C 1-6 is alkyl; R 8b are the same or different and are long chain alkyl; R 8c are the same or different and represent the general formulas [9A] to [9E]: [ka] (In the formula, * is the bond position; and R 9 is a long chain alkyl and / or long chain alkenyl) Substituents represented by Selected from ; R 8d are the same or different and each represent a hydrogen atom, a halogen atom, a long-chain alkyl optionally substituted with 1 to 13 halogen atoms, or a long-chain alkyloxy optionally substituted with 1 to 13 halogen atoms; R 8e teeth (1) long-chain alkyl, (2) long-chain alkyl-carbonyl, or (3) benzoyl substituted with 1 to 5 long chain alkyloxy and / or long chain alkenyloxy; and R 8f teeth, (1) long-chain alkyl, (2) long-chain alkyl-carbonyl, or (3) Long-chain alkenyl-carbonyl Substituents represented by Selected from ;and L is a group represented by the general formula

[10] : [ka] (In the formula, * is the bonding position to Z; ** is the bond position to T; and L 1 may be substituted C 2-10Alkylene or optionally substituted C 6-10 It is an arylene ) , or general formula [10-1]: [ka] (In the formula, * is the bonding position to Z; ** is the bond position to T; and L 1 may be substituted C 1-10 Alkylene or optionally substituted C 6-10 arylene) is a substituent represented by is a substituent represented by the formula: T is a single bond or a group of the general formula

[11] : [ka] (In the formula, X and W are as defined above; *teeth 、O is the bonding position with; ** is the bond position to G; and q is an integer from 0 to 10. where G is a silicon substituent, T is a single bond. and a compound of formula [B-1]: [ka] [In the formula, B P , W, and X are as defined above, and p is an integer of 1 to 50, preferably an integer of 1 to 30, more preferably an integer of 1 to 25, and most preferably an integer of 1 to 15; L' is OH, -O - N + H (aliphatic amine), -O - N + H (cyclic amine), or -O - N + H (aromatic amine), preferably -O - N+ H(C 1-6 alkyl)3 or -O - N + H (cyclic amine) and -O - N + H(C 1-6 Alkyl)3 is, for example, -O - N + H(CH2CH3)3, -O - N + H (cyclic amine) is, for example, -O - (HDBU) + and; Q 1 is a group that can be removed under acidic conditions] and a compound of (1) treating with at least one condensing agent selected from the group consisting of phosphorus reagent 1, phosphorus reagent 2, and onium reagent in the presence of a base; and (2) Treat with an oxidizing agent and an organic amine Formula [C-1]: [ka] [In the formula, B P , Q 1 , W, X, G, T, n, and p are as defined above. A method for obtaining the compound of

[0012] <2> Furthermore, formula [C-1]: [ka] [In the formula, B P , Q 1 , W, X, G, T, n, and p are as defined above. with an acid, or an acid and a scavenger, to Formula [C-0-1]: [ka] [In the formula, B P , Q 1 , W, X, G, T, n, and p are as defined above. compound of, and The compound of formula [C-0-1] and a compound of formula [B-0-1]: [ka] [In the formula, B P , Q 1 , W, and L′ are as defined above. and a compound of (1) treating with at least one condensing agent selected from the group consisting of phosphorus reagent 1, phosphorus reagent 2, and onium reagent in the presence of a base, and then (2) Treat with an oxidizing agent and an organic amine Formula [C-1-1]: [ka] [In the formula, B P , Q 1 , W, X, G, T, n, and p are as defined above. obtaining a compound of formula <1> The method described below.

[0013] <3> Furthermore, formula [C-1]: [ka] [In the formula, B P , Q 1 , W, X, G, T, n, and p are as defined above. The compound is treated under conditions for removing a protecting group of a hydroxyl group, Formula [C-0-2]: [ka] [In the formula, B P , Q 1 , W, X, n, and p are as defined above. to obtain a compound of (1) Formula [P-1]: [ka] [Wherein W is an oxygen atom or a sulfur atom, and R 1and R 2 are the same or different and each represents H, optionally substituted C 1-6 alkyl, optionally substituted phenyl, and PH(=O)-OH] and a base, and subsequently (2) Treat with a hydrolysis solution Formula [C-0-2-1]: [ka] [In the formula, B P , Q 1 , W, X, L′, n, and p are as defined above. obtaining a compound of formula <1> The method described below.

[0014] <4> Furthermore, the formula [C-0-2-1]: [ka] [In the formula, B P , Q 1 , W, X, L′, n, and p are as defined above. and a compound of formula [A-0-1]: [ka] [In the formula, B P , G, and T are as defined above. and a compound of (1) treating with at least one condensing agent selected from the group consisting of phosphorus reagent 1, phosphorus reagent 2, and onium reagent in the presence of a base, and then (2) Treat with an oxidizing agent and an organic amine Formula [C-1-2]: [ka] [In the formula, B P , Q 1 , W, X, G, T, n, and p are as defined above. obtaining a compound of formula <3> The method described below.

[0015] <5> Furthermore, the formula [B-1-0]: [ka] [In the formula, B P , Q 1 , W, X, and p are as defined above. The compound (1) Formula [P-1]: [ka] [In the formula, W is an oxygen atom or a sulfur atom , R 1 , and R 2 teeth, each of which may be the same or different, is H, optionally substituted C 1-6 selected from the group consisting of alkyl, optionally substituted phenyl, and PH(=O)-OH ] and a base, and subsequently (2) Treat with a hydrolysis solution Formula [B-1]: [ka] [In the formula, B P , Q 1 , W, X, L′, and p are as defined above. obtaining a compound of formula <1> The method described below.

[0016] <6> Furthermore, the formula [A-0]: [ka] [In the formula, B P , Q 1 , W, X, G, T, and n are as defined above. with an acid, or an acid and a scavenger, to Formula [A-1]: [ka] [In the formula, B P, W, X, G, T, and n are as defined above. obtaining a compound of formula <1> The method described below.

[0017] <7> G is a compound having the formula: TIFF0007825613000028.tif115140 [In the formula, * represents the bonding position to T.] selected from the group consisting of: T is a single bond; <1> ~ <6> A method according to any one of the preceding claims.

[0018] <8> In formula [B-1], B P are the same or different and are optionally protected nucleobases, Q 1 is trityl or dimethoxytrityl, L' is -O - N + H(CH2CH3)3, or -O - (HDBU) + and W is O, and p is an integer of 2 to 20, preferably an integer of 2 to 15, and more preferably an integer of 2 to 10; <1> The compound of formula [B-1] described in

[0019] <9> ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[C Bz -A Bz ]―ON), Triethylammonium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[C Bz -A Bz ]-ON (triethylamine salt) ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphazene (6-benzamido-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[C Bz -A Bz -TC Bz -T]-ON), Triethylammonium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methyl (6-benzamido-9H-purin-9-yl)morpholin-2-yl)methoxy)phosphoryl)-6-(6-benzamido-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[C Bz -A Bz -TC Bz -T]-ON (triethylamine salt) ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-((((2S,6R)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)-4-tritylmorpholine- 2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[C Bz -G CE,Pac -A Bz -A Bz -G CE,Pac ]-ON), Triethylammonium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-((((2S,6R)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)-4-trityl morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[C Bz -G CE,Pac -A Bz -A Bz -G CE,Pac ]-ON (triethylamine salt) ((2S,6R)-4-((((2S,6R)-4-((((2S,6R)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3, 4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[G CE,Pac -G CE,Pac -TTC Bz ]-ON), Triethylammonium ((2S,6R)-4-((((2S,6R)-4-((((2S,6R)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl) ... 2-(2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[G CE,Pac -G CE,Pac -TTC Bz ]-ON (triethylamine salt) ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[C Bz -C Bz ]-ON), Triethylammonium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[C Bz -C Bz ]-ON (triethylamine salt) ((2S,6R)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[TT]-ON), Triethylammonium ( (2S,6R)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[TT]-ON (triethylamine salt)), ((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[TTT]-ON), triethylammonium ((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[TTT]-ON(triethylamine salt)), ((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)((((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-di hydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[TTTT]-ON), Triethylammonium ((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)((((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl hydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[TTTT]-ON (triethylamine salt)), 1,8-Diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpho purin-2-yl)methoxy)phosphoryl)morpholin-2-yl)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)phosphoryl)-6-(6-benzamido-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate (i.e., H-phosphonate-PMO[C Bz -A Bz -TC Bz -T]-ON (DBU salt), and 1,8-Diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-4-(((2S,6R)-4-(((2S,6R)-6-(6-benzamidopurin-9-yl)-4-(((2S,6R)-6-(6-benzamidopurin-9-yl)-4-(((2S,6R)-4-(((2S,6R)-6-(6-(2-cyanoethoxy)-2- ((2-phenoxyacetyl)amino)purin-9-yl)-4-(((2S,6R)-4-(((2S,6R)-6-(6-(2-cyanoethoxy)-2-((2-phenoxyacetyl)amino)purin-9-yl-4-(dimethylamino-(((2S,6R)-4-(dimethylamino-(((2S,6R)-6-(5-methyl-2,4-dioxo- pyrimidine -1-yl)-4-trityl-morpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-pyrimidin-1-yl)morpholin-2-yl)methoxy)phosphoryl)morpholin-2-yl)methoxy-(dimethylamino)phosphoryl-6-(5-methyl-2,4-dioxo-pyrimidin-1-yl)morpholin-2-yl)methoxy-(dimethylamino)phosphoryl)morpholin-2-ylmethoxy-(dimethylamino)phosphoryl)-6-(6-(3-cyanopropoxy-2-((2-phenoxyacetyl)amino)phosphoryl)

[0023] H-phosphonate-PMO[TG-10-yl]-6-(6-(2-cyanoethoxy)-2-((2-phenoxyacetyl)amino)purin-9-yl)morpholin-2-yl)methoxy-(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-pyrimidin-1-yl)morpholin-2-yl)methoxyphosphinate (i.e., ... CE,Pac -A Bz -A Bz -G CE,Pac -G CE,Pac -TG CE,Pac -TT]-ON (DBU salt) <8> The compound of formula [B-1] described in

[0020] <10> In formula [B-0-1], B P is an optionally protected nucleobase, Q 1 is trityl or dimethoxytrityl, L' is O - (HDBU) + and W is O, <2> The compound of formula [B-0-1] described in

[0021] <11> 1,8-diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methylphosphonate, 1,8-diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methylphosphonate, 1,8-diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methylphosphonate, 1,8-diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methylphosphonate, and 1,8-Diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-(2-(2-phenylacetamido)-6-((4-(pivaloyloxy)benzyl)oxy)-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methylphosphonate selected from the group consisting of <10> The compound of formula [B-0-1] described in [Effects of the Invention]

[0022] Oligonucleic acid compounds are compounds having a structure in which two or more nucleoside units are linked via phosphorus bonds. To produce an oligonucleic acid compound, it is necessary to perform condensation reactions multiple times to form phosphorus bonds between adjacent nucleoside units. According to the present invention, phosphorus bonds can be formed efficiently, which is expected to result in a reduction in the time required to produce an oligonucleic acid compound. DETAILED DESCRIPTION OF THE INVENTION

[0023] The present invention will be described in detail below.

[0024] Terminology Examples of "nucleobases" include adenine, guanine, hypoxanthine, cytosine, thymine, uracil, and modified bases thereof. Examples of such modified bases include pseudouracil, 3-methyluracil, dihydrouracil, 5-alkylcytosine (e.g., 5-methylcytosine), 5-alkyluracil (e.g., 5-ethyluracil), 5-halouracil (5-bromouracil), 6-azapyrimidine, 6-alkylpyrimidine (6-methyluracil), 2-thiouracil, 4-thiouracil, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, 5'-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyluracil, 1-methyladenine, 1-methylhypoxanthine, 2,2-dimethylguanine, 3-methylcytosine, 2-methyladenine, 2-methylguanine, N 6 -methyladenine, 7-methylguanine, 5-methoxyaminomethyl-2-thiouracil, 5-methylaminomethyluracil, 5-methylcarbonylmethyluracil, 5-methyloxyuracil, 5-methyl-2-thiouracil, 2-methylthio-N 6 -isopentenyladenine, uracil-5-oxyacetic acid, 2-thiocytosine, purine, 2,6-diaminopurine, 2-aminopurine, isoguanine, indole, imidazole, and xanthine, but are not limited to these. PThe amino group or hydroxyl group of the nucleic acid base may be protected. "Optionally protected nucleobase" encompasses both unprotected and protected nucleobases, such as adenine, guanine, hypoxanthine, cytosine, thymine, uracil, and the like, in which the amino and / or hydroxyl groups are unprotected or protected. The amino-protecting group is not particularly limited as long as it is used as a protecting group for nucleic acids, and specific examples thereof include benzoyl, 4-methoxybenzoyl, acetyl, propionyl, butyryl, isobutyryl, phenylacetyl, phenoxyacetyl, 4-tert-butylphenoxyacetyl, 4-isopropylphenoxyacetyl, and (dimethylamino)methylene. Preferred amino-protecting groups are benzoyl, acetyl, phenylacetyl, and 4-tert-butylphenoxyacetyl. Examples of hydroxyl-protecting groups include 2-cyanoethyl, 4-nitrophenethyl, phenylsulfonylethyl, methylsulfonylethyl, trimethylsilylethyl, phenyl optionally substituted with 1 to 5 electron-withdrawing groups at any substitutable position, diphenylcarbamoyl, dimethylcarbamoyl, diethylcarbamoyl, methylphenylcarbamoyl, 1-pyrrolidinylcarbamoyl, morpholinocarbamoyl, 4-(tert-butylcarboxy)benzyl, 4-[(dimethylamino)carboxy]benzyl, and 4-(phenylcarboxy)benzyl (see, for example, International Publication No. 2009 / 064471A1). Preferred hydroxyl-protecting groups include 2-cyanoethyl, 4-nitrophenethyl, and 4-(tert-butylcarboxy)benzyl. Preferred protecting groups for the 6-hydroxyl group of guanine include 2-cyanoethyl. In one embodiment, protected nucleobases include, for example, those shown below: [ka] [wherein Pg represents a protecting group]. A more specific embodiment of the protected nucleic acid base is adenine (A) in which the amino group is protected with benzoyl.Bz ), cytosine with the amino group protected by benzoyl (C Bz ), guanine (G) in which the hydroxyl group is protected with 2-cyanoethyl and the amino group is protected with phenoxyacetyl. CE,Pac ) and the like, but are not limited to these. The term "long-chain alkyl" refers to, for example, a linear or branched alkyl having 10 to 300 carbon atoms, preferably a linear or branched alkyl having 10 to 100 carbon atoms, and more preferably a linear or branched alkyl having 10 to 30 carbon atoms. The "long-chain alkyl" portion of "long-chain alkyl-carbonyl" and "long-chain alkyloxy" can be the same as the "long-chain alkyl" defined above. "Long-chain alkenyl" refers to, for example, a linear or branched alkenyl having 10 to 300 carbon atoms, preferably a linear or branched alkenyl having 10 to 100 carbon atoms, and more preferably a linear or branched alkenyl having 10 to 30 carbon atoms. The "long-chain alkenyl" portion of "long-chain alkenyloxy" and "long-chain alkenyl-carbonyl" can be the same as the "long-chain alkenyl" described above. Examples of "halogen" include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom. Examples of the "5- or 6-membered saturated cyclic amino" include a 5- or 6-membered saturated cyclic amino group having one or two N atoms, which may have one O or S atom as a ring-constituting atom, and specific examples thereof include 1-pyrrolidinyl, 1-imidazolidinyl, piperidino, 1-piperazinyl, 1-tetrahydropyrimidinyl, 4-morpholino, 4-thiomorpholino, 1-homopiperazinyl, and oxazolidin-3-yl. "C 1-6 The term "alkyl" refers to a straight-chain or branched alkyl having 1 to 6 carbon atoms, and specific examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl. "C 1-6The term "alkoxy" refers to a straight-chain or branched-chain alkoxy having 1 to 6 carbon atoms, and specific examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentyloxy, and n-hexyloxy. "C 1-6 Alkoxy-C 1-6 "C" in "Alkyl" 1-6 The "alkoxy" portion is the "C 1-6 The same as "alkoxy" can be mentioned. "Ji (C 1-6 alkyl)amino”, mono(amino-C substituted with a group removable under basic conditions 1-6 Alkyl)amino, di(amino-C substituted with a group removable under basic conditions 1-6 Alkyl)amino, mono(C 1-6 Alkyl)amino-C 1-6 Alkyl, di(C 1-6 Alkyl)amino-C 1-6 Alkyl, Tri(C 1-6 Alkyl)ammonio-C 1-6 Alkyl, mono(C 1-6 alkyl)amino, di(C 1-6 alkyl)amino, tri(C 1-6 Alkyl)ammonio, mono(amino-C 1-6 Alkyl)amino and di(amino-C 1-6 Alkyl)amino "C 1-6 The "C alkyl" portion is 1-6 The same can be mentioned as "alkyl". "C 2-10 "Alkylene" is a divalent group formed by removing one hydrogen atom bonded to a different constituent carbon atom from a straight-chain or branched-chain alkyl having 2 to 10 carbon atoms, and examples thereof include ethylene, propylene, isopropylene, butylene, pentylene, and hexylene. Such "alkylene" may be substituted with 1 to 12 halogen atoms at any substitutable position. L 1 As the "alkylene" in the above, ethylene is particularly preferred. "C6-10 "Arylene" is a divalent group formed by removing two hydrogen atoms bonded to two different ring carbon atoms from a monocyclic or polycyclic aromatic hydrocarbon having 6 to 10 carbon atoms, and examples thereof include phenylene and naphthylene. Such "arylene" may be substituted with 1 to 6 halogen atoms at any substitutable position. L 1 As the "arylene" in the above, phenylene is particularly preferred. "1,1,3,3-tetra(C 1-6 alkyl)guanidyl", "C 1-6 Alkoxy-C 1-6 Alkyl" "Di(C 1-6 alkyl)amino", "di(C 1-6 Alkyl)amino-C 1-6 Alkyl, Tri(C 1-6 alkyl)ammonio", "tri(C 1-6 Alkyl)ammonio-C 1-6 alkyl," "mono(C) substituted with a group that can be removed under basic conditions" 1-6 alkyl)amino," "mono(C substituted with a group that can be removed under basic conditions" 1-6 Alkyl)amino-C 1-6 alkyl," "mono (amino-C substituted with a group that can be removed under basic conditions) 1-6 alkyl)amino," "di(amino-C substituted with a group that can be removed under basic conditions" 1-6 (Alkyl)amino" 1-6 The "C alkyl" portion is 1-6 The same can be mentioned as "alkyl". Examples of the "group removable under acidic conditions" include trityl, monomethoxytrityl, tert-butyldimethylsilyl, and dimethoxytrityl. An example of the "group removable under basic conditions" is trifluoroacetyl. Examples of the "group removable under neutral conditions" include groups that can be removed by the action of tetrabutylammonium fluoride or hydrogen trifluoride-triethylamine salt, such as 2-cyanoethoxymethoxy, 2-cyanoethoxy-2-ethoxy, and tert-butyldimethylsilyl. Examples of the "silicon substituent" include triphenylsilyl, diisopropylphenylsilyl, tert-butyldimethylsilyl, and tert-butyldiphenylsilyl. "Aryl" can include, for example, phenyl. Examples of "heteroaryl" include pyridyl, pyrimidyl, pyridazyl, pyrazinyl, thienyl, and furanyl. Generally, any "solid phase support" that can be used in the solid phase synthesis of nucleic acids, peptides, peptide nucleic acids, sugars, etc. can be used without any particular problems. Examples of such a "solid phase support" include controlled pore glass (CPG), oxalated controlled pore glass (see, for example, Nucleic Acids Research, Vol. 19, 1527 (1991)), TentaGel support-aminopolyethylene glycol derivatized support (see, for example, Tetrahedron Letters, Vol. 34, 3373 (1993)), Poros-polystyrene / divinylbenzene copolymer, polystyrene resin, and polyacrylamide resin. Examples of "soluble polymers that dissolve in organic solvents" include non-crosslinked styrene polymers and polyethylene glycol derivatives. The "soluble polymer soluble in an organic solvent" portion of "(soluble polymer soluble in an organic solvent)-oxy" and "(soluble polymer soluble in an organic solvent)-amino" can be the same as the "soluble polymer soluble in an organic solvent" described above. Examples of "non-crosslinked styrene polymers" include derivatives of polystyrene that are not crosslinked with divinylbenzene and have a spacer such as polyethylene glycol (TentaGel series, ArgoGel series). Examples of "polyethylene glycol derivatives" include derivatives of polyethylene glycol having a molecular weight of 100 to 40,000 and having a substituent (SUNBRIGHT (registered trademark) series).

[0025] <Extension reaction> One aspect of the present invention is a method for obtaining a compound of formula [C-1] by reacting a compound of formula [A-1] with a compound of formula [B-1]: [ka] [In the formula, B P , Q 1 , W, X, G, T, n, L′, and p are as defined above. and Step (1): Condensation and Step (2): Oxidative amination may include:

[0026] One aspect of the present invention is Formula [A-1]: [ka] [In the formula, B P ,W,X,G,T and n is , as defined above] and a compound of formula [B-1]: [ka] [In the formula, B P , Q 1 , W, X, L′, and p are as defined above. and a compound of Step (1): Treating with at least one condensing agent selected from the group consisting of phosphorus reagent 1, phosphorus reagent 2, and onium reagent in the presence of a base, and Step (2): Treating with an oxidizing agent and an organic amine Formula [C-1]: [ka] [In the formula, B P , Q 1 , W, X, G, T, n, and p are as defined above. This is a way to obtain

[0027] <Step (1): Condensation>

[0028] <Condensing Agent for Step (1)> In one embodiment of the present invention, the condensing agent can be at least one selected from the group consisting of phosphorus reagent 1, phosphorus reagent 2, and onium reagent.

[0029] <Specific examples of condensing agents in step (1)> In one embodiment of the present invention, the condensing agent may be at least one selected from the group consisting of phosphorus reagent 1, phosphorus reagent 2, and onium reagents, and examples thereof include diphenyl chlorophosphate, bis(2,6-dimethylphenyl) chlorophosphate, bis(2-oxo-3-oxazolidinyl)phosphinic chloride, PyBrop (bromotripyrrolidinophosphonium hexafluorophosphate), PyBOP ((benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate), PyClop (chlorotripyrrolidinophosphonium hexafluorophosphate), PyAOP ((7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate), PyNTP (3-nitro-1,2,4-triazol-1-yl-tris(pyrrolidin-1-yl)phosphonium hexafluorophosphate), HATU (1-[bis(dimethylamino)methylene]-1H -1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate), COMU ((1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate), HBTU (1-[bis(dimethylamino)methylene]-1H-benzotriazolium 3-oxide hexafluorophosphate), BOI (2-(benzotriazol-1-yl)oxy-1,3-dimethylamino) and 1-(1',3'-dimethyl-1H-imidazol-2'-yl)-3-nitro-1,2,4-triazole hexafluorophosphate), DMINTP (1-(1',3'-dimethyl-1H-imidazol-2'-yl)-3-nitro-1,2,4-triazole hexafluorophosphate), and more preferably diphenyl chlorophosphate, bis(2,6-dimethylphenyl) chlorophosphate, PyBrop, or BOI, and more preferably diphenyl chlorophosphate, bis(2,6-dimethylphenyl) chlorophosphate, or PyBrop.

[0030] In one embodiment of the present invention, the phosphorus reagent 1 is Formula [P-2]: [ka] [In the formula, W is an oxygen atom or a sulfur atom; R 1 and R 2 are the same or different and are selected from the group consisting of an optionally substituted aromatic ring -O- and an optionally substituted heterocycle, preferably Phenoxy , 2,6-dimethyl Phenoxy and 2-oxooxazolidides] The compound is preferably one selected from diphenyl chlorophosphate, bis(2,6-dimethylphenyl) chlorophosphate, and bis(2-oxo-3-oxazolidinyl)phosphinic chloride, and more preferably diphenyl chlorophosphate or bis(2,6-dimethylphenyl) chlorophosphate.

[0031] In one embodiment of the present invention, the phosphorus reagent 2 is Formula [P-3]: [ka] [In the formula, R 1 , R 2 , R 3 , and R 4 are the same or different and may be halogen, C 1-6 Alkylamino, di(C 1-6 alkyl)amino, optionally substituted cyclic C 1-6 selected from the group consisting of amino, optionally substituted aromatic amino, optionally substituted aromatic ring, and optionally substituted heterocycle-O—; or R 1 and R 2 together with the phosphorus atom to which they are attached form an optionally substituted ring structure, and R 3 and R 4 are the same or different and may be halogen, C 1-6 Alkylamino, di(C 1-6 alkyl)amino, optionally substituted cyclic C 1-6selected from the group consisting of amino, optionally substituted aromatic amino, optionally substituted aromatic ring, and optionally substituted heterocycle-O—; Preferably, R 1 , R 2 , and R 3 is an optionally substituted cyclic C 1-6 amino, and R 4 is halogen, optionally substituted aromatic amino, or optionally substituted heterocycle-O—; More preferably, R 1 , R 2 , and R 3 is pyrrolidyl, and R 4 is chloro, bromo, benzotriazol-1-yloxy, 7-azabenzotriazol-1-yloxy, or 3-nitro-1,2,4-triazol-1-yl], preferably one selected from PyBrop, pyBOP, PyClop, PyAOP, and PyNTP, and more preferably PyBrop.

[0032] In one aspect of the invention, the onium reagent is Formula [O]: [ka] [In the formula, R 1 , R 2 and R 3 teeth , the same or different, halogen, C 1-6 Alkylamino, di(C 1-6 alkyl)amino, optionally substituted cyclic C 1-6 Amino, optionally substituted aromatic amino, optionally substituted aromatic ring, optionally substituted heterocycle-O—, and optionally substituted C 1-6 alkylideneaminooxy; or R 1 and R 2together with the carbon atoms to which they are attached form a ring structure, and R 3 is a halogen, C 1-6 Alkylamino, di(C 1-6 alkyl)amino, optionally substituted cyclic C 1-6 selected from the group consisting of amino, optionally substituted aromatic amino, optionally substituted aromatic ring, and optionally substituted heterocycle-O—; Preferably, R 1 and R 2 are the same or different, 1-6 alkyl)amino and optionally substituted cyclic C 1-6 amino, and R 3 is an optionally substituted heterocycle-O- or an optionally substituted C 1-6 alkylideneaminooxy; or R 1 and R 2 are taken together with the carbon atoms to which they are attached to form a ring structure and are optionally substituted aromatic amino; More preferably, R 1 and R 2 is dimethylamino, and R 3 is O-(7-azabenzotriazol-1-yl) or O-(benzotriazol-1-yl), or R 1 is dimethylamino, and R 2 is a morpholino, and R 3 is 1-cyano-2-ethoxy-2-oxoethylideneaminooxy], preferably one selected from HATU, COMU, HBTU, BOI, DMINTP, more preferably is B OI in be.

[0033] <Base in Step (1)> In one embodiment of the present invention, the base is at least one selected from the group consisting of primary amines, secondary amines, tertiary amines, cyclic amines, and aromatic amines, and is, for example, diisopropylethylamine, triethylamine, N-ethylmorpholine, pyridine, lutidine, collidine, N-methylimidazole, N,N-dimethylimidazole, or 1-benzylimidazole, preferably diisopropylethylamine, pyridine, N-methylimidazole, or 1-benzylimidazole, and more preferably pyridine or N-methylimidazole.

[0034] <Combination of condensing agent and base in step (1)> In one embodiment of the present invention, the combination of the condensing agent and the base is not particularly limited, and examples thereof include diphenyl chlorophosphate and pyridine, diphenyl chlorophosphate and N-methylimidazole, bis(2,6-dimethylphenyl) chlorophosphate and pyridine, bis(2,6-dimethylphenyl) chlorophosphate and N-methylimidazole, PyBroP and pyridine, PyBroP and N-methylimidazole, PyBroP and diisopropylethylamine, and BOI and diisopropylethylamine, and preferably diphenyl chlorophosphate and pyridine, diphenyl chlorophosphate and and N-methylimidazole, bis(2,6-dimethylphenyl) chlorophosphate and pyridine, bis(2,6-dimethylphenyl) chlorophosphate and N-methylimidazole, PyBroP and pyridine, PyBroP and N-methylimidazole, PyBroP and diisopropylethylamine, and more preferably diphenyl chlorophosphate and pyridine, diphenyl chlorophosphate and N-methylimidazole, bis(2,6-dimethylphenyl) chlorophosphate and N-methylimidazole, PyBroP and N-methylimidazole, PyBroP and diisopropylethylamine.

[0035] <Amount of Condensing Agent Used in Step (1)> In one embodiment of the present invention, the condensing agent can be used in an amount of, for example, 0.5 to 100 molar equivalents, preferably 1 to 50 molar equivalents, and more preferably 2 to 20 molar equivalents relative to the compound of formula [B-1].

[0036] <Amount of base used in step (1)> In one embodiment of the present invention, the base can be used in an amount of, for example, 1 to 100 molar equivalents, preferably 2 to 50 molar equivalents, and more preferably 2 to 40 molar equivalents, relative to the compound of formula [A-1].

[0037] <Reaction solvent for step (1)> In one embodiment of the present invention, the solvent used in step (1) is not particularly limited, and for example, at least one selected from the group consisting of dichloromethane, chloroform, 1,2-dichloroethane, tetrahydrofuran, toluene, acetonitrile, and ethyl acetate can be used.

[0038] <Reaction temperature in step (1)> In one embodiment of the present invention, the reaction temperature in the step (1) is not particularly limited, and is, for example, -10 to 80°C, preferably 0 to 60°C, and more preferably 10 to 40°C.

[0039] <Reaction time of step (1)> In one embodiment of the present invention, the reaction time in the step (1) is not particularly limited, and is, for example, 1 minute to 12 hours, preferably 2 minutes to 4 hours, and more preferably 5 minutes to 2 hours.

[0040] <(2): Oxidative amination>

[0041] <Oxidizing agent for step (2)> In one embodiment of the present invention, the oxidizing agent is not particularly limited as long as it can oxidize a phosphorus atom, and examples thereof include iodine and carbon tetrachloride. Preferably, the oxidizing agent is a dichloromethane solution of iodine, a chloroform solution of iodine, or a tetrahydrofuran solution of iodine, and more preferably, a tetrahydrofuran solution of iodine.

[0042] <Amount of oxidizing agent used in step (2)> In one aspect of the present invention, oxidizing agent can be used in an amount of, for example, 0.5 to 100 molar equivalents, preferably 1 to 50 molar equivalents, and more preferably 2 to 10 molar equivalents relative to the compound of formula [B-1].

[0043] <Organic amine in step (2)> In one embodiment of the present invention, the organic amine is not particularly limited and is, for example, at least one selected from the group consisting of primary amines, secondary amines, tertiary amines, cyclic amines, and aromatic amines, such as dimethylamine, diisopropylamine, piperidine, pyrrolidine, N-methylpiperazine, or morpholine, preferably dimethylamine, diisopropylamine, piperidine, or pyrrolidine, and more preferably dimethylamine.

[0044] <Amount of organic amine used in step (2)> In one embodiment of the present invention, the organic amine can be used in an amount of, for example, 1 to 200 molar equivalents, preferably 2 to 100 molar equivalents, and more preferably 2 to 50 molar equivalents relative to the compound of formula [B-1].

[0045] <Reaction solvent for step (2)> In one embodiment of the present invention, the solvent used in step (2) is not particularly limited, and for example, at least one selected from the group consisting of dichloromethane, chloroform, 1,2-dichloroethane, tetrahydrofuran, toluene, acetonitrile, and ethyl acetate can be used.

[0046] <Reaction temperature in step (2)> In one embodiment of the present invention, the reaction temperature in the step (2) is not particularly limited, and is, for example, -10 to 80°C, preferably 0 to 60°C, and more preferably 0 to 40°C.

[0047] <Reaction time of step (2)> In one embodiment of the present invention, (2) The reaction time is not particularly limited and is, for example, 1 minute to 4 hours, preferably 2 minutes to 2 hours, and more preferably 2 minutes to 1 hour.

[0048] <Compound of formula [A-1]>

[0049] <Synthesis of Compound of Formula [A-1]> One aspect of the present invention is a method for obtaining a compound of formula [A-1] from a compound of formula [A-1-0]. [ka] [In the formula, B P , Q 1 , W, X, G and T is , as defined above Yes and n is an integer of 1 to 50, preferably an integer of 1 to 30, more preferably an integer of 1 to 25, and most preferably an integer of 1 to 15.

[0050] The compounds of formula [A-1-0] may be synthesized using methods known in the art.

[0051] <Compound of formula [B-1]>

[0052] One aspect of the present invention is a compound represented by the formula [B-1], wherein p=1: [ka] Compounds of the formula (I) include, for example, compounds listed in Table 1 below.

[0053] [Table 1]

[0054] <Method for obtaining the compound of formula [B-1]> One aspect of the present invention is a method for obtaining a compound of formula [B-1] from a compound of formula [B-1-0]: [ka] [In the formula, B P , Q 1 , W, X, L′, and p are as defined above. and Step (1): Phosphonation Step (2): Hydrolysis, and Optionally, step (3): ion exchange may include:

[0055] The compound of formula [B-1-0] may be synthesized using methods known in the art.

[0056] Specific examples of the compound of formula [B-1-0] where p=1 include the compounds listed in Table 3 below.

[0057] [Table 2]

[0058] <Step (1): Phosphonation> One aspect of the present invention is Formula [B-1-0]: [ka] [In the formula, B P , Q 1 , W, X, and p are as defined above. The compound Formula [P-1]: [ka] [Wherein W is an oxygen atom or a sulfur atom, and R 1 and R 2 are the same or different and represent H, optionally substituted C 1-6 alkyl, optionally substituted phenyl, PH(=O)-OH] and a base, Formula [B-1'-0]: [ka] [In the formula, B P , Q 1 , W, X, R 1 and p have the same meanings as above. The method includes a step of obtaining a compound of the formula:

[0059] <Phosphonylating Reagent for Step (1)> In one embodiment of the present invention, a compound represented by formula [P-1]: [ka] wherein W is as defined above and R 1 and R 2 are the same or different and represent H, optionally substituted C 1-6 alkyl, optionally substituted phenyl, PH(=O)-OH, preferably R 1 is H, optionally substituted C 1-6 alkyl, or optionally substituted phenyl; R 2 is optionally substituted C 1-6 alkyl, or optionally substituted phenyl, or PH(=O)-OH, more preferably R 1 and R 2 is phenyl. is preferably diphenyl phosphite, phosphorous acid, bis(1,1,1,3,3,3-hexafluoropropan-2-yl)phosphonate, or diphosphonic acid, more preferably diphenyl phosphite.

[0060] <Base in Step (1)> In one embodiment of the present invention, the base is at least one selected from the group consisting of primary amines, secondary amines, tertiary amines, cyclic amines, and aromatic amines, and is, for example, N-ethylmorpholine, pyridine, triethylamine, or N-methylimidazole, preferably N-ethylmorpholine or pyridine, and more preferably N-ethylmorpholine.

[0061] <Combination of phosphonylation reagent and base in step (1)> In one embodiment of the present invention, the combination of the phosphonylation reagent and the base is not particularly limited, and examples thereof include diphenyl phosphite and N-ethylmorpholine, diphenyl phosphite and pyridine, diphenyl phosphite and N-methylimidazole, and phosphorous acid and pyridine, preferably diphenyl phosphite and N-ethylmorpholine, diphenyl phosphite and pyridine, and more preferably diphenyl phosphite and N-ethylmorpholine.

[0062] <Amount of phosphonylation reagent used in step (1)> In one embodiment of the present invention, the compound of formula [P-1] can be used in an amount of, for example, 1 to 100 molar equivalents, preferably 1 to 40 molar equivalents, and more preferably 2 to 20 molar equivalents relative to the compound of formula [B-1-0].

[0063] <Amount of base used in step (1)> In one embodiment of the present invention, the base can be used in an amount of, for example, 1 to 100 molar equivalents, preferably 1 to 40 molar equivalents, and more preferably 4 to 40 molar equivalents, relative to the compound of formula [B-1-0].

[0064] <Reaction solvent for step (1)> In one embodiment of the present invention, the solvent used in step (1) is not particularly limited, and for example, at least one selected from the group consisting of dichloromethane, chloroform, 1,2-dichloroethane, tetrahydrofuran, toluene, and ethyl acetate can be used.

[0065] <Reaction temperature in step (1)> In one embodiment of the present invention, the reaction temperature in the step (1) is not particularly limited, and is, for example, -10 to 60°C, preferably 0 to 50°C, and more preferably 20 to 50°C.

[0066] <Reaction time of step (1)> In one embodiment of the present invention, the reaction time in the step (1) is not particularly limited, and is, for example, 5 minutes to 24 hours, preferably 10 minutes to 18 hours, and more preferably 30 minutes to 12 hours.

[0067] <Step (2): Hydrolysis> One aspect of the present invention is Formula [B-1'-0]: [ka] [In the formula, B P , Q 1 , W, X, R 1 and p have the same meanings as above. The compound Treated with a hydrolysis solution, Formula [B-1]: [ka] [In the formula, B P , Q 1 , W, X, and p are as defined above, and L' is OH, -O - N + H (aliphatic amine), -O - N + H (cyclic amine), or -O - N + H (aromatic amines)] The method includes a step of obtaining a compound of formula (I). In one embodiment of the present invention, in formula [B-1], B P , Q 1 , W, X, and p are as defined above; L' is -O - N + H (aliphatic amine), -O - N+ H (cyclic amine), or -O - N + H (aromatic amine), the compound of formula [B-1] , fat The compound is an aliphatic amine salt, a cyclic amine salt, or an aromatic amine salt, for example, a compound of the formula [B-1] [wherein B P , Q 1 , W, X, and p are as defined above, and L' is -OH] is a salt formed with an aliphatic amine, a cyclic amine, or an aromatic amine. In one embodiment of the present invention, in formula [B-1], B P , Q 1 , W, X, and p are as defined above, and L' is preferably -O - N + H(C 1-6 alkyl)3 or -O - N + H (cyclic amine) and -O - N + H(C 1-6 Alkyl)3 is, for example, -O - N + H(CH2CH3)3, -O - N + H (cyclic amine) is, for example, -O - (HDBU) + and DBU is 1,8-diazabicyclo[5.4.0]undec-7-ene.

[0068] <Hydrolysis solution of step (2)> In one embodiment of the present invention, the hydrolysis solution is an aqueous solution containing at least one base selected from the group consisting of aliphatic amines (primary amines, secondary amines, and tertiary amines), cyclic amines, and aromatic amines. Examples of aliphatic amines include trimethylamine, triethylamine, and diisopropylethylamine. Examples of cyclic amines include diazabicycloundecene (DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene), piperidine, piperazine, morpholine, and N-methylmorpholine. Examples of aromatic amines include pyridine, imidazole, and N-methylimidazole.

[0069] In one embodiment of the present invention, the hydrolysis solution is, for example, an aqueous solution containing at least one base selected from the group consisting of triethylamine, diazabicycloundecene, N-methylimidazole, and N-methylmorpholine, preferably an aqueous solution in which the base and carbon dioxide are dissolved, and more preferably an aqueous solution of triethylammonium hydrogencarbonate or diazabicycloundecene hydrogencarbonate.

[0070] <Amount of hydrolysis solution used in step (2)> In one embodiment of the present invention, the hydrolysis solution can be used in an amount of, for example, 1 to 100 molar equivalents, preferably 1 to 50 molar equivalents, and more preferably 2 to 20 molar equivalents relative to the compound of formula [B-1'-0].

[0071] <Reaction solvent for step (2)> In one embodiment of the present invention, the solvent used in step (2) is not particularly limited, and for example, at least one selected from the group consisting of dichloromethane, chloroform, 1,2-dichloroethane, tetrahydrofuran, toluene, acetonitrile, and ethyl acetate can be used.

[0072] <Reaction temperature in step (2)> In one embodiment of the present invention, the reaction temperature in the step (2) is not particularly limited, and is, for example, -10 to 60°C, preferably 0 to 40°C, and more preferably 0 to 30°C.

[0073] <Reaction time of step (2)> In one embodiment of the present invention, the reaction time in the step (2) is not particularly limited, and is, for example, 1 minute to 24 hours, preferably 10 minutes to 12 hours, and more preferably 10 minutes to 2 hours.

[0074] <Step (3): Ion exchange> One aspect of the present invention is Formula [B-1'-0]: [ka] [In the formula, B P , Q 1 , W, X, R 1 and p have the same meanings as above. is treated with a hydrolysis solution to Formula [B-1']: [ka] [In the formula, B P , Q 1 , W, X, and p are as defined above, and L″ is O - N + H(C 1-6 alkyl)3] The compound is further treated with a hydrolysis solution to Formula [B-1]: [ka] [In the formula, B P , Q 1 , W, X, and p are as defined above, and L' is -O - N + H (cyclic amine) and -O - N + H (cyclic amine) is, for example, -O - (HDBU) + is] The method includes a step of obtaining a compound of formula (I).

[0075] <Hydrolysis solution of step (3)> In one embodiment of the present invention, the hydrolysis solution is, for example, an aqueous solution containing a cyclic amine, preferably an aqueous solution in which a cyclic amine and carbon dioxide are dissolved, and more preferably an aqueous solution of diazabicycloundecene hydrogencarbonate.

[0076] <Amount of hydrolysis solution used in step (3)> In one embodiment of the present invention, the hydrolysis solution can be used in an amount of, for example, 1 to 50 molar equivalents, preferably 1 to 50 molar equivalents, and more preferably 2 to 20 molar equivalents relative to the compound of formula [B-1'-0] or the compound of formula [B-1'].

[0077] <Reaction solvent for step (3)> In one embodiment of the present invention, the solvent used in step (3) is not particularly limited, and for example, at least one selected from the group consisting of dichloromethane, chloroform, 1,2-dichloroethane, tetrahydrofuran, toluene, and ethyl acetate can be used.

[0078] <Reaction temperature in step (3)> In one embodiment of the present invention, the reaction temperature in the step (3) is not particularly limited, and is, for example, -10 to 60°C, preferably 0 to 40°C, and more preferably 0 to 30°C.

[0079] <Reaction time in step (3)> In one embodiment of the present invention, the reaction time in the step (3) is not particularly limited, and is, for example, 1 minute to 24 hours, preferably 10 minutes to 12 hours, and more preferably 10 minutes to 2 hours.

[0080] <Q in the molecule of compound [C-1] 1 How to remove > One aspect of the present invention is to obtain a compound of formula [C-1], Q 1 to obtain a compound of formula [C-0-1]: [ka] [In the formula, B P , W, X, G, T, n, and p are as defined above, and Q 1 is a group that can be removed under acidic conditions, and is preferably trityl, monomethoxytrityl, or dimethoxytrityl, more preferably trityl or dimethoxytrityl, and most preferably trityl.

[0081] One aspect of the present invention is a compound represented by formula [C-1]: [ka] [In the formula, B P , Q 1 , W, X, G, T, n, and p are as defined above. with an acid alone or with an acid and a scavenger, Formula [C-0-1]: [ka] [In the formula, B P , W, X, G, T, n, and p are as defined above. The method includes a step of obtaining a compound of formula (I).

[0082] <The above acid: Q 1 Removal Reagents> In one embodiment of the present invention, the acid is not particularly limited as long as it is an acid commonly used in the technical field, and examples thereof include trifluoroacetic acid, cyanopyridine trifluoroacetate, triethylamine trifluoroacetate, cyanoacetic acid, trichloroacetic acid, phosphoric acid, methanesulfonic acid, p-toluenesulfonic acid, and hydrochloric acid. When using these acids, they may be combined with a base (e.g., triethylamine) to adjust the acidity.

[0083] In one embodiment of the present invention, the acid may be, for example, a commercially available deblocking solution for nucleic acid synthesis [e.g., Deblocking Solution - 1.3 w / v% trichloroacetic acid / dichloromethane solution (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.), Deblocking Mix 3% dichloroacetic acid / dichloromethane solution (manufactured by Glen Research)].

[0084] <The above acid: Q 1 Amount of removal reagent used> In one embodiment of the present invention, the acid can be used in an amount of, for example, 1 to 500 molar equivalents, preferably 2 to 200 molar equivalents, and more preferably 2 to 50 molar equivalents relative to the compound of formula [C-1].

[0085] <The above acid: Q 1 Removal reagent concentration > In one embodiment of the present invention, the acid is suitably diluted with a reaction solvent to a concentration within a range of, for example, 1% to 80%, and preferably diluted to a concentration within a range of 2% to 50%.

[0086] <The above Q 1 Removal of reaction solvent> In one aspect of the present invention, 1 The solvent used in the removal step is not particularly limited as long as it is a solvent commonly used in the art, and a single solvent may be used, or two or more solvents may be mixed and used. In one aspect of the present invention, 1 Examples of the solvent used in the removal step include aromatic solvents such as benzene, toluene, xylene, and mesitylene; ester solvents such as ethyl acetate and isopropyl acetate; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, and cyclohexane; and halogenated solvents. These solvents may be used in combination. In one aspect of the present invention, 1 The solvent used in the removal step is, for example, chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixed solvent thereof, and is preferably one selected from the group consisting of chloroform, dichloromethane, and 2,2,2-trifluoroethanol.

[0087] <The aforementioned Scavenger:Q 1 Removal Reagents> ​​​In one embodiment of the present invention, the scavenger may be, for example, ethanol, triisopropylsilane, 1-hydroxybenzotriazole, pyrrole, indole, 2,2,2-trifluoroethanol, methanol, anisole, mercaptoethanol, or thioanisole, and is preferably one selected from the group consisting of ethanol, triisopropylsilane, and 2,2,2-trifluoroethanol.

[0088] <The aforementioned Scavenger:Q 1 Amount of removal reagent used> In one embodiment of the present invention, the scavenger can be used in an amount of, for example, 1 to 100 molar equivalents, preferably 1 to 50 molar equivalents, and more preferably 1 to 20 molar equivalents relative to the compound of formula [C-1].

[0089] <Q 1 Removal reaction temperature In one aspect of the present invention, 1 The reaction temperature for the removal step is not particularly limited, and is, for example, -10 to 40°C, preferably 0 to 30°C, and more preferably 0 to 25°C.

[0090] <Q 1 Removal reaction time> In one aspect of the present invention, 1 The reaction time for the removal step is not particularly limited, and is, for example, 1 minute to 24 hours, preferably 10 minutes to 6 hours, and more preferably 10 minutes to 2 hours.

[0091] In one embodiment of the present invention, in the method for producing compound [C-0-1], a solution containing an acid is added to a reaction mixture containing compound [C-1] produced by subjecting compound [A-1] and compound [B-1] to a condensation reaction, thereby obtaining Q 1 The elimination reactions of (1) and (2) can be carried out in the same system as this continuous reaction.

[0092] ​​In one embodiment of the present invention, the solvent that can be used in the continuous reaction is not particularly limited as long as it is a solvent that is commonly used in the technical field, and a single solvent may be used, or two or more solvents may be mixed and used.

[0093] In one embodiment of the present invention, examples of solvents that can be used in this continuous reaction include aromatic solvents such as benzene, toluene, xylene, and mesitylene; ester solvents such as ethyl acetate and isopropyl acetate; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, and cyclohexane; halogenated solvents; etc. These solvents may be used in combination.

[0094] One aspect of the present invention is a compound of formula [A-0], 1 to obtain a compound of formula [A-1]: [ka] [In the formula, B P , W, X, G, T, n, and p are as defined above, Q 1 is a group that can be removed under acidic conditions, and is preferably trityl, monomethoxytrityl, or dimethoxytrityl, more preferably trityl or dimethoxytrityl, and most preferably trityl.

[0095] One aspect of the present invention is a compound of formula [A-0]: [ka] [In the formula, B P , Q 1 , W, X, G, T, and n are as defined above. with an acid alone or with an acid and a scavenger, Formula [A-1]: [ka] [In the formula, B P , W, X, G, T, and n are as defined above. The method includes a step of obtaining a compound of formula (I).

[0096] In one embodiment of the present invention, Q is obtained from the compound of formula [A-0] 1 The step of obtaining the compound of formula [A-1] by removing Q 1 The same "acid," "amount of acid used," "scavenger," "amount of scavenger used," "reaction solvent," "reaction time," and "reaction temperature" as those described in "Method for removing the acid," "scavenger," "reaction solvent," "reaction time," and "reaction temperature" can be used.

[0097] <Removal of hydroxyl protecting groups> One aspect of the present invention is a method for obtaining a compound of formula [C-0-2] by removing GT from a compound of formula [C-1]: [ka] [In the formula, B P , Q 1 , W, X, G, T, n, and p are as defined above. Includes:

[0098] One aspect of the present invention is a compound represented by formula [C-1]: [ka] [In the formula, B P , Q 1 , W, X, and p are as defined above; G is (1) silicon substituents, (2) optionally substituted C 1-18 Alkyl-carbonyl, optionally substituted C 1-18 alkoxy-carbonyl, long chain alkyl-carbonyl, or long chain alkoxy-carbonyl; (3) Benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; or (4) The following general formula [7]: [ka] (In the formula, * indicates the binding position with T; Z is (1) (soluble polymer soluble in organic solvent)-oxy, (2) (Soluble polymer soluble in organic solvent)-amino, (3) long-chain alkyloxy, benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or benzyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; (4) a solid support, or (5) The following general formulas [8A] to [8N]: [ka] (In the formula, * indicates the bonding position with L; j represents an integer from 0 to 4; k represents an integer from 0 to 5; R 8a is a hydrogen atom or C 1-6 represents alkyl; R 8b are the same or different and each represents a long-chain alkyl; R 8c are the same or different and are represented by the following general formulas [9A] to [9E]: [ka] (In the formula, * represents a bond position; and R 9 represents a long chain alkyl and / or a long chain alkenyl. represents a substituent represented by the formula: R 8d are the same or different and represent a hydrogen atom, a halogen atom, a long-chain alkyl optionally substituted with 1 to 13 halogen atoms, or a long-chain alkyloxy optionally substituted with 1 to 13 halogen atoms; R 8e teeth (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; and R 8f teeth, (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) represents a long-chain alkenyl-carbonyl. represents a substituent represented by the formula: L is the general formula

[10] : [ka] (In the formula, * indicates the bonding position with Z; ** represents the bond position with T; and L 1 is optionally substituted C 2-10 Alkylene or optionally substituted C 6-10 Represents an arylene or general formula [10-1]: [ka] (In the formula, * indicates the bonding position with Z; ** represents the bond position with T; and L 1 is optionally substituted C 1-10 Alkylene or optionally substituted C 6-10 represents arylene.) represents a substituent represented by represents a substituent represented by the formula: n represents an integer from 1 to 50; and T is a single bond or a group of the following general formula

[11] : [ka] (In the formula, X and W are as defined above; * represents the bonding position with **O, *O, or *N in the above general formulas [4a] to [4d]; ** represents the bond position with G; and q represents an integer from 0 to 10. However, when G is a silicon substituent, T is a single bond. The compound is treated under conditions for removing a protecting group of a hydroxyl group, Formula [C-0-2]: [ka] [In the formula, B P , Q 1 , W, X, n, and p are as defined above. The method includes a step of obtaining a compound of formula (I).

[0099] In one embodiment of the present invention, a compound represented by formula [C-1]: [ka] [In the formula, B P , Q 1 , W, X, and p are as defined above; G is (1) silicon substituents, (2) optionally substituted C 1-18 Alkyl-carbonyl, optionally substituted C 1-18 alkoxy-carbonyl, long chain alkyl-carbonyl, or long chain alkoxy-carbonyl, or (3) Benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy groups; n represents an integer from 1 to 50; and T is a single bond. is a compound of

[0100] In one embodiment of the present invention, a compound represented by formula [C-1]: [ka] [In the formula, B P , Q 1 , W, X, and p are as defined above; G is (4) The following general formula [7]: [ka] (In the formula, * indicates the binding position with T; Z is (3) long-chain alkyloxy, benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or benzyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; (4) a solid support, or (5) The following general formulas [8A], [8B], [8E], and [8F]: [ka] (In the formula, * indicates the bonding position with L; R 8a is a hydrogen atom or C 1-6 represents alkyl; R 8b are the same or different and each represents a long-chain alkyl; R 8e teeth (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) Benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy. represents a substituent represented by the formula: L is the general formula

[10] : [ka] (In the formula, * indicates the bonding position with Z; ** represents the bond position with T; and L 1 is optionally substituted C 2-10 Alkylene or optionally substituted C 6-10 Represents an arylene or general formula [10-1]: [ka] (In the formula, * indicates the bonding position with Z; ** represents the bond position with T; and L 1 is optionally substituted C 1-10 Alkylene or optionally substituted C 6-10 represents arylene.) represents a substituent represented by represents a substituent represented by the formula: n represents an integer from 1 to 50; and T is a single bond or a group of the following general formula

[11] : [ka] (In the formula, X and W are as defined above; * represents the bonding position with **O, *O, or *N in the above general formulas [4a] to [4d]; ** represents the bond position with G; and q represents an integer from 0 to 10. However, when G is a silicon substituent, T is a single bond. is a compound of

[0101] In one embodiment of the present invention, a compound represented by formula [C-1]: [ka] [In the formula, B P , Q 1 , W, X, n, and p are as defined above; G has the following formula: [ka] [In the formula, * represents the bonding position to T; T is a single bond. is a compound of

[0102] <Conditions for removing the hydroxyl-protecting group> In one embodiment of the present invention, the conditions for removing the hydroxyl-protecting group can be selected depending on the type or properties of the protecting group in the compound of formula [C-1], and the compound of formula [C-0-2] can be obtained by removing the hydroxyl-protecting group.

[0103] In one embodiment of the present invention, the conditions for removing the hydroxyl-protecting group are, for example, the conditions for deprotecting the hydroxyl-protecting group described in "Green's PROTECTIVE GROUPS in ORGANIC SYNTHESIS, 4th Edition, 2006".

[0104] In one embodiment of the present invention, the hydroxyl-protecting group is (1) a silicon substituent, and T is a single bond, and the hydroxyl-protecting group can be removed by treatment with, for example, tetrabutylammonium fluoride.

[0105] In one embodiment of the present invention, the protecting group for the hydroxyl group is (2) optionally substituted C 1-18 Alkyl-carbonyl, optionally substituted C 1-18 an alkoxy-carbonyl, a long chain alkyl-carbonyl, or a long chain alkoxy-carbonyl; (3) Benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy groups, and T is a single bond. The hydroxyl-protecting group can be removed by treatment with sodium methoxide, for example.

[0106] In one embodiment of the invention, G is (4) The general formula [7]: [ka] (wherein Z, L, and * are as defined above) and T is a single bond, the hydroxyl-protecting group can be removed by treatment with, for example, (1) aqueous ammonia, (2) aqueous ammonia / ethanol, or (3) a mixed solution of aqueous ammonia and aqueous methylamine.

[0107] <Conditions for removing the hydroxyl protecting group: amount of reagent used> In one embodiment of the present invention, the reagent can be used in an amount of, for example, 1 to 200 molar equivalents, preferably 2 to 100 molar equivalents, and more preferably 2 to 50 molar equivalents relative to the compound of formula [C-1].

[0108] <Conditions for removing the hydroxyl protecting group: Reaction solvent> In one embodiment of the present invention, the solvent used in the <Removal of the hydroxyl-protecting group> is not particularly limited as long as it is a solvent commonly used in the technical field, and a single solvent may be used, or two or more solvents may be mixed and used. In one embodiment of the present invention, examples of the solvent used in the <removal of the hydroxyl-protecting group> include aromatic solvents such as benzene, toluene, xylene, and mesitylene; ester solvents such as ethyl acetate and isopropyl acetate; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, and cyclohexane; halogenated solvents; etc. These solvents may be used in combination.

[0109] <Conditions for removing the hydroxyl protecting group: Reaction temperature> In one embodiment of the present invention, the reaction temperature for the <removal of the hydroxyl-protecting group> is not particularly limited, and is, for example, 0 to 80°C, preferably 10 to 60°C, and more preferably 20 to 40°C.

[0110] <Conditions for removing the hydroxyl protecting group: Reaction time> In one embodiment of the present invention, the water Acid group protecting group The reaction time for the removal step is not particularly limited, and is, for example, 10 minutes to 24 hours, preferably 10 minutes to 12 hours, and more preferably 30 minutes to 6 hours.

[0111] <Final deprotection and nucleic acid compound isolation process> When compound [C-1] has protecting groups in its molecule, a deprotection treatment appropriate for the type or nature of the protecting groups can be performed to produce a compound from which all protecting groups have been removed. For example, all protecting groups in the compound can be removed according to the deprotection method described in "Green's Protective Groups in Organic Synthesis, 4th Edition, 2006." Specifically, the protecting groups for the substituents [6] and amino or hydroxyl groups of the nucleic acid bases in the compound [C-1] molecule can be removed by treating the compound with, for example, (1) aqueous ammonia, (2) aqueous ammonia / ethanol, or (3) a mixed solution of aqueous ammonia and methylamine. In addition, for example, the Q 1 The group removable under acidic conditions substituted on the protecting group of the amino group at the 3'-position of the nucleoside on the 3'-terminal side of compound [C-1] can be removed by treating with the same "acid" as described in "Method for removing the amino group" or a solution of hydrochloric acid or acetic acid diluted in an appropriate solvent.

[0112] <About the purification and separation process> The compound [C-1] from which all the protecting groups have been removed can be separated and purified from the reaction mixture by conventional separation and purification means, for example, extraction, concentration, neutralization, filtration, centrifugation, recrystallization, and separation of C8 to C 18 The protein can be isolated by using, alone or in combination, means such as reverse phase column chromatography, cation exchange column chromatography, anion exchange column chromatography, gel filtration column chromatography, high performance liquid chromatography, dialysis, and ultrafiltration (see, for example, International Publication No. 1991 / 09033A1). When the desired compound is purified using reverse phase chromatography, a mixed solution of 20 mM triethylamine / acetic acid buffer and acetonitrile can be used as the elution solvent, for example. Also, when purifying a desired compound using ion exchange chromatography, for example, a mixed solution of 1 M saline and 10 mM aqueous sodium hydroxide or 0.3 M saline of 50 mM phosphate buffer can be used.

Example

[0113] The present invention will be described in more detail below with reference examples, reference examples, comparative examples, and test examples, but the present invention is not limited thereto. Compound names were generated using software (ChemDraw or BIOVIA Draw). Note that the "conversion yield (%)" means the ratio of the raw material converted to the target product, and is calculated by "{peak area (%) corresponding to the target product detected by high performance liquid chromatography (hereinafter referred to as "HPLC")} ÷ {peak area (%) corresponding to the raw material detected by HPLC + peak area (%) corresponding to the target product detected by HPLC} × 100". HPLC conditions: 0.5 mg of the product was dissolved in acetonitrile or 20% water / acetonitrile, and HPLC analysis was performed under the following conditions. The conversion efficiency was calculated by using the integrated value of the peak area obtained by absorption at UV = 264 nm in HPLC. <ODS condition 1> Column: Waters XBridge C18 (2.5 μm, 4.6 × 50 mm), 60 °C Detection wavelength: 264 nm [[ID=2I]]Mobile phase A: 20 mM AcO NH 4aq. Mobile phase B: MeCN Flow rate: 0.75 mL / min Gradient: 40 - 95% B (0 - 15 min) 95% B (15 - 24 min) 40% B (24 - 30 min) <ODS condition 2> Column: Waters XBridge C8 (5 μm, 4.6 × 75 mm), 60 °C Detection wavelength: 264 nm Mobile phase A: 50 mM TEAAaq Mobile phase B: MeOH Flow rate: 0.75 mL / min Gradient: 70 - 95% B (0 - 20 min), 95% B (20 - 26 min), 75% B (26 - 35 min) <LC / MS Condition 1> Equipment used: ACQUITY UPLC ultra-high speed liquid chromatograph (waters) Quadrupole time-of-flight mass spectrometer SYNAPT-MS (waters) Column: ACQUITY UPLC BEH C18 1.7 μm, 2.1×50 mm (waters) Temperature: 50 °C Flow rate: 0.2 mL / min Mobile phase A: 10 mM aqueous ammonia Mobile phase B: MeCN Gradient: 50 - 95% B (4 min) Detector 1: UV264 nm Detector 2: Quadrupole time-of-flight mass spectrometer Ionization method: ESI+ Measurement range: 100 - 2000 m / z <LC / MS Condition 2> Equipment used: ACQUITY UPLC ultra-high speed liquid chromatograph (waters) Quadrupole time-of-flight mass spectrometer SYNAPT-MS (waters) Column: ACQUITY UPLC BEH C18 1.7 μm, 2.1×50 mm (waters) Temperature: 50 °C Flow rate: 0.2 mL / min Mobile phase A: 10 mM aqueous ammonia Mobile phase B: MeOH Gradient: 70 - 95% B (4 min) Detector 1: UV264 nm Detector 2: Quadrupole time-of-flight mass spectrometer Ionization method: ESI+ Measurement range: 100 - 2000 m / z <LC / MS Condition 3> Equipment used: Agilent 6100 series single quadrupole LC / MS system (Agilent) Column:Waters XBridge C18(3.5μm,4.6×150mm) Temperature: 35℃ Flow rate: 1.0mL / min Mobile phase A: 20 mM ammonium acetate aqueous solution Mobile phase B: MeCN Gradient: 20-99%B (15 min) Detector 1: UV 264 nm Detector 2: Quadrupole time-of-flight mass spectrometer Ionization method: ESI+ Measurement range: 250-3000 m / z

[0114] Reference example 1 4-(octadecylamino)-4-oxobutanoate [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl (hereinafter referred to as "G1-suc-morT-OFF")

[0115] Step 1: Preparation of 4-(octadecylamino)-4-oxobutanoic acid (hereinafter referred to as "G1-suc") To a solution of octadecane-1-amine (21.94 g) in dichloromethane (500 mL), succinic anhydride (8.96 g, 1.1 eq) and triethylamine (17 mL, 1.5 eq) were added and stirred at room temperature for 7 hours. The mixture was concentrated under reduced pressure, and 150 mL of acetone was added to the residue, followed by stirring for 16 hours. The precipitate was filtered with suction, washed with acetone (400 mL), and then dried under reduced pressure at 30°C for 3 hours to obtain G1-sucrose (29.1 g, 96.6%) as a white powder.

[0116] Step 2: Preparation of [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl 4-(octadecylamino)-4-oxobutanoate (hereinafter referred to as "G1-suc-morT-OFF") To a solution of G1-suc (14.4 g) in tetrahydrofuran (150 mL), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (8.56 g, 1.2 eq.) was added and stirred at room temperature. Subsequently, 1-((2R,6S)-6-(hydroxymethyl)-4-tritylmorpholin-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (hereinafter referred to as "morT-OH") (18 g, 1.0 eq.) and 4.57 g of 4-(N,N-dimethylamino)pyridine were added and stirred in a water bath at 70°C for 1 hour. After cooling to room temperature, 0.1 M aqueous sodium dihydrogen phosphate was added and stirred for a while. The aqueous layer was removed, and the organic layer was washed once with 0.1 M aqueous sodium dihydrogen phosphate and once with saturated brine diluted twice with water. The combined aqueous layers were extracted with dichloromethane, and the combined organic layers were dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated and the residue was dried under reduced pressure to give 4-(octadecylamino)-4-oxobutanoate [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methyl (hereinafter referred to as "G1-suc-morT-ON") (white amorphous, 28.1 g, 89.5%). This was dissolved in 140 mL of dichloromethane and stirred in an ice bath. 20 mL of 2,2,2-trifluoroethanol and 10.3 mL of triisopropylsilane were added. After stirring for a while, 5.1 mL of trifluoroacetic acid was added dropwise. One hour after the dropwise addition, the reaction mixture was poured into 100 mL of saturated aqueous sodium bicarbonate solution with ice while stirring. After confirming that the aqueous layer had a pH of 7-8, the aqueous layer was extracted with dichloromethane. The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by column chromatography on silica gel using a dichloromethane-methanol mixture as the mobile phase, and dried under reduced pressure to obtain 19.89 g of G1-suc-morT-OFF as a powder.

[0117] 1H-NMR(CDCl3):δ8.90(1H,bs);7.25(1H,d,J=1.6Hz);5.72(1H,dd,J=9.6Hz,2.6Hz);5 .65(1H,m);4.14(2H,d,J=5.2Hz);3.98(1H,m);3.23(2H,dd,J=12.8Hz,7.0Hz);3.12(2 H,dd,J=12Hz,2.6Hz);2.95(2H,dd,J=12.8Hz,1.8Hz);2.60~2.75(4H,m);2.47(2H,t,J =6.8Hz);1.95(3H,d,J=1.6Hz);1.48(2H,m), 1.21~1.34(29H,m);0.88(3H,t,J=6.4Hz) ESI-MS(+):593.36(M+H)(LC / MS conditions 1)

[0118] Reference example 1-1 In the same way as in Reference Example 1, (1) 4-(octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl]methyl ester (G1-suc-morA-ON), 4-(octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(6-benzamido-9H-purin-9-yl)morpholin-2-yl]methyl ester (G1-suc-morA-OFF) (2) <4-(octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methyl> (G1-suc-morC-ON), <4-(octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)morpholin-2-yl]methyl> (G1-suc-morC-OFF) (3) <4-(octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(6-(2-cyanoethoxy)-2-(phenoxyacetamido)-9H-purin-9-yl)-4-tritylmorpholin-2-yl]methyl> (G1-suc-morG-ON), <4-(octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(6-(2-cyanoethoxy)-2-(phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl]methyl> (G1-suc-morG-OFF) was manufactured.

[0119] Reference Example 2: Succinic acid {[(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1-yl)morpholin-2-yl]methyl}{2-octadecanoyloxy-1-[(octadecanoyloxymethyl)ethyl]} (hereinafter referred to as "G2-suc-morT-OFF").

[0120] Step 1: Preparation of 4-((1,3-bis(stearoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (hereinafter referred to as "G2-suc") Dichloromethane (8 mL) was added to 1 g (1.60 mmol) of 2-hydroxypropane-1,3-diyl distearate, followed by 176 mg (1.76 mmol) of succinic anhydride and 293 mg (2.40 mmol) of 4-(N,N-dimethylamino)pyridine, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, 1 M aqueous sodium dihydrogen phosphate solution was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated to give G2-sucrose (1.40 g).

[0121] Step 2: Preparation of succinic acid {[(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1-yl)-4-trityl-morpholin-2-yl]methyl}{2-octadecanoyloxy-1-[(octadecanoyloxymethyl)ethyl]} (hereinafter referred to as "G2-suc-morT-ON") Dichloromethane (5.2 mL) was added to G2-suc (900 mg, 1.24 mmol) and 277 mg (1.45 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, followed by 500 mg (1.03 mmol) of morT-OH and 132 mg (1.09 mmol) of 4-(N,N-dimethylamino)pyridine. The mixture was stirred at room temperature for 16 hours. After the reaction was complete, 0.1 M aqueous sodium dihydrogen phosphate was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give G2-suc-morT-ON (1.09 g, 89%).

[0122] 1H-NMR(CDCl3):δ8.04(1H,s);7.17~7.51(15H,m);6.98(1H,s);6.12(1H,dd,J=9.6H z,2.4Hz);5.25(1H,m);4.34~4.37(1H,m);4.26~4.30(2H,m);4.11~4.16(2H,m);4. 00~4.08(2H,m);3.35(1H,d,J=11.2Hz);3.10(1H,d,J=11.6Hz);2.60(4H,s);2.30( 4H,t,J=7.6Hz);1.83(3H,s);1.38~1.44(2H,m);1.24(60H,m);0.87(6H,t,J=6.8Hz)

[0123] Process 3 Manufacturing of G2-suc-morT-OFF Dichloromethane (4.2 mL) was added to G2-suc-morT-ON and stirred at 0°C. Next, 127 μL (0.62 mmol) of triisopropylsilane and 64 μL (0.82 mmol) of trifluoroacetic acid were added at 0°C and stirred at room temperature for 1 hour. After the reaction was completed, saturated aqueous sodium bicarbonate solution was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give G2-suc-morT-OFF (373 mg, 95%).

[0124] 1 H-NMR(CDCl3):δ8.04(1H,bs);7.24(1H,s);5.70(1H,d,J=2Hz);5.21~5.26(1H,m);4.28~4.31(2H,m);4.13~4.17(4H,m);3.96~4.00(1H,m);3.1 1(1H,dd,J=12.4,2Hz);2.94(1H,dd,J=12.8,2.4Hz);2.57~2.65(6H,m);2.32(4H,t,J=7.6Hz);1.95(3H,s);1.25(60H,m);0.88(6H,t,J=7.6Hz)

[0125] Reference Example 3 1,3-bis(oleoyloxy)propan-2-yl [{(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl]succinate (hereinafter referred to as "G3-suc-morT-OFF")

[0126] Step 1: Preparation of 1,3-bis(oleoyloxy)propan-2-yl [{(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl]succinate (hereinafter referred to as "G3-suc-morT-ON") 4-((1,3-bis(oleoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (hereinafter referred to as "G3-suc") was produced using 2-hydroxypropane-1,3-diyl dioleate as a starting material in the same manner as in Step 1 of Reference Example 2. G3-suc-morT-ON was then produced in the same manner as in Step 2 of Reference Example 2.

[0127] 1 H-NMR(CDCl3):δ8.00(1H,s);7.17~7.51(15H,m);6.99(1H,s);6.09~6.12(1H,m);5.29~5.3 8(4H,m);5.20~5.25(1H,m);4.33~4.37(1H,m);4.26~4.30(2H,m);4.12~4.16(2H,m);4.00~ 4.09(2H,m);3.35(1H,d,J=11.6Hz);2.15(1H,d,J=11.6Hz);2.60(4H,m);2.30(4H,t,J=7.2 Hz);1.97~2.02(8H,m);1.83(3H,s);1.57~1.61(2H,m);1.28(44H,m);0.89(6H,t,J=6.8Hz)

[0128] Process 2 Manufacturing of G3-suc-morT-OFF It was produced in the same manner as in Step 3 of Reference Example 2.

[0129] 1 H-NMR(CDCl3): δ7.97(1H,bs);7.24(1H,s);5.69~5.72(1H,m);5.29~5.38(4 H,m);5.21~5.25(1H,m);4.27~4.31(2H,m);4.13~4.17(4H,m);3.97~3.99(1 H,m);3.11(1H,d,J=12Hz);2.94(1H,d,J=13.2Hz);2.57~2.67(4H,m)2.31(4 H,t,J=7.6Hz);1.99~2.00(11H,m);1.26~1.29(46H,m);0.87(6H,t,J=6.8Hz)

[0130] Reference Example 4 {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl 4-oxo-4-(4-stearoylpiperazin-1-yl)butanoate (hereinafter referred to as "G4-suc-morT-OFF") Step 1: Production of 4-oxo-4-(4-stearoylpiperazin-1-yl)butanoic acid (hereinafter referred to as "G4-suc") To 1.68 g (5.91 mmol) of stearic acid, 1.13 g (5.91 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 0.79 g (5.91 mmol) of 1-hydroxybenzotriazole, 26 mL of tetrahydrofuran was added. Then, 1.45 mL (10.7 mmol) of triethylamine and 1 g (5.37 mmol) of tert-butyl piperazine-1-carboxylate were added and stirred at room temperature for 16 hours. After completion of the reaction, saturated aqueous sodium bicarbonate was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give tert-butyl 4-stearoylpiperazine-1-carboxylate (1.64 g; 67%). 18 mL of dichloromethane was added to the mixture and stirred at 0 °C. 2.77 mL (36.2 mmol) of trifluoroacetic acid was added at 0 °C and stirred at room temperature for 2 hours. After the reaction was complete, saturated aqueous sodium bicarbonate was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated to give 1-(piperazin-1-yl)octadecan-1-one (1.30 g). 18 mL of dichloromethane was added to 1.3 g (3.70 mmol) of the crude product, followed by 0.41 g (4.10 mmol) of succinic anhydride and 0.77 mL (5.50 mmol) of triethylamine, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was evaporated, and acetone was added to the residue, followed by slurry washing at room temperature for 16 hours. The insoluble matter was collected by suction filtration, washed with acetone, and dried to give G4-suc (1.20 g).

[0131] Step 2: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl 4-oxo-4-(4-stearoylpiperazin-1-yl)butanoate (hereinafter referred to as "G4-suc-morT-ON") Tetrahydrofuran (10 mL) was added to G4-suc (982 mg, 2.17 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (555 mg, 2.90 mmol) and stirred at 70 °C. Next, morT-OH (1 g, 2.07 mmol) and 4-(N,N-dimethylamino)pyridine (265 mg, 2.17 mmol) were added and stirred at 70 °C for 30 minutes. After the reaction was complete, the mixture was allowed to cool to room temperature. 0.1 M aqueous sodium dihydrogen phosphate solution was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give G4-suc-morT-ON (1.68 g, 89%).

[0132] 1 H-NMR(CDCl3):δ8.00(1H,s);7.16~7.50(15H,m);6.97(1H,s);6.10(1H,d ,J=8Hz);4.34~4.36(1H,m);4.04(2H,d,J=4.8Hz);3.57~3.64(4H,m);3.4 4~3.48(4H,m);3.32~3.34(1H,m);3.09~3.12(1H,m);2.60~2.64(4H,m);2 .31(2H,t,J=7.6Hz);1.82(3H,s);1.23~1.42(32H,m);0.86(3H,t,J=6.8)

[0133] Process 3 Manufacturing of G4-suc-morT-OFF It was produced in the same manner as in step 3 of Reference Example 2.

[0134] 1 H-NMR(CDCl3):δ8.32(1H,bs);7.23(1H,s);5.67~5.70(1H,m);4.12~4.19(2H,m);3.96~4.01(1H,m);3.47~3.67(8H,m)3.10~3 .13(1H,m);2.95~2.98(1H,m);2.60~2.72(4H,m);2.33(2H,t,J=7.2Hz);1.95(3H,s);1.25~1.31(32H,m);0.88(3H,t,J=7.6Hz)

[0135] Reference Example 5 [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl 4-(octadecylcarbamoyl)benzoate (hereinafter referred to as "G5-tpa-morT-OFF") Step 1: Preparation of [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methyl 4-(octadecylcarbamoyl)benzoate (hereinafter referred to as "G5-tpa-morT-ON") This was prepared in the same manner as in Step 2 of Reference Example 2, using 4-(octadecylcarbamoyl)benzoic acid.

[0136] 1 H-NMR(CDCl3):δ8.24(1H,s);7.97(2H,d,J=8Hz);7.77(2H,d,J=8Hz);7.17~7.46(15H,m);6.95(1H,s);6.12~6.16(1H,m);4.49~4.51(1 H,m);4.25~4.33(2H,m);3.42~3.47(2H,m);3.35~3.38(1H,m);3.21~3.24(1H,m);1.79(3H,s);1.23~1.44(34H,m);0.86(3H,t,J=6.8Hz)

[0137] Step 2: Production of G5-tpa-morT-OFF It was produced in the same manner as in step 3 of Reference Example 2.

[0138] 1 H-NMR(CDCl3):δ8.24(1H,bs);8.11(2H,d,J=8.4Hz);7.84(2H,d,J=8.4Hz);7.24(1H,s);6.14~6.17(1H,m);5.74~5.77(1H,m);4.40~4.45 (2H,m);4.13~4.19(1H,m);3.45~3.50(2H,m);3.14~3.18(1H,m);3.05~3.08(1H,m);1.93(3H,s);1.26~1.41(34H,m);0.89(3H,t,J=7.6Hz)

[0139] Reference Example 6 {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl 4-(4-(4-(octadecylcarbamoyl)benzoyl)piperazin-1-yl)-4-oxobutanoate (hereinafter referred to as "G6-suc-morT-OFF"). Step 1: Preparation of 4-(4-(4-(octadecylcarbamoyl)benzoyl)piperazin-1-yl)-4-oxobutanoic acid (hereinafter referred to as "G6-suc") This was produced in the same manner as in Step 1 of Reference Example 4, except that 4-(octadecylcarbamoyl)benzoic acid was used instead of stearic acid.

[0140] Step 2: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl 4-[4-{4-(octadecylcarbamoyl)benzoyl}piperazin-1-yl]-4-oxobutanoate (hereinafter referred to as "G6-suc-morT-ON") It was produced in the same manner as in step 2 of Reference Example 2.

[0141] 1 H-NMR(CDCl3):δ8.08(1H,bs);7.81(2H,d,J=7.6Hz);7.16~7.50(17H,m);6.97(1H,s);6.08~6.10(1H,m);4.33~4.39(1H,m);4.0 2~4.04(2H,m);3.31~3.79(11H,m);3.08~3.11(1H,m);2.60~2.69(4H,m);1.81(3H,s);1.23~1.44(34H,m);0.86(3H,t,J=6.4Hz)

[0142] Process 3 Manufacturing of G6-suc-morT-OFF To 493 mg (0.47 mmol) of G6-suc-morT-ON, 4.6 mL of dichloromethane and 0.4 mL of 2,2,2-trifluoroethanol were added and stirred at 0°C. Next, 145 μL (0.70 mmol) of triisopropylsilane and 53 μL (0.70 mmol) of trifluoroacetic acid were added at 0°C and stirred at room temperature for 1 hour. After completion of the reaction, saturated aqueous sodium bicarbonate solution was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give G6-suc-morT-OFF (372 mg; 98%).

[0143] 1 H-NMR(CDCl3):δ8.05(1H,bs);7.79(2H,d,J=7.6Hz);7.45(2H,d,J=7.6Hz );7.23(1H,s);6.08~6.11(1H,m);5.67~5.69(1H,m);4.10~4.15(2H,m);3 .96~3.99(1H,m);3.36~3.79(8H,m);3.08~3.11(1H,m);2.93~2.96(1H,m) ;2.57~2.70(6H,m);1.92(3H,s);1.23~1.38(34H,m);0.86(3H,t,J=7.2Hz)

[0144] Reference Example 7 {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl 3,4,5-tris(octadecyloxy)benzoate (hereinafter referred to as "G7-morT-OFF")

[0145] Step 1: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-trityl-morpholin-2-yl}methyl 3,4,5-tris(octadecyloxy)benzoate (hereinafter referred to as "G7-morT-ON") This was produced in the same manner as in Step 2 of Reference Example 2, using 3,4,5-trioctadecoxybenzoic acid.

[0146] 1 H-NMR(CDCl3):δ7.90(1H,bs);7.12~7.45(17H,m);6.97(1H,s);6.12~6.14(1H,m);4.46~4.51(1H,m);4.28~4.32(1H,m);4.16~4 .20(1H,m);3.90~4.00(6H,m);3.37~3.40(1H,m);3.22~3.25(1H,m);1.78~1.82(5H,m);1.23~1.50(96H,m);0.86(9H,t,J=6.8Hz)

[0147] Step 2: Production of G7-morT-OFF It was produced in the same manner as in step 3 of Reference Example 2.

[0148] 1 H-NMR(CDCl3): δ7.98(1H,bs);7.22(3H,m);5.69~5.72(1H,m);4.32~4.36(2H,m);4.08~4.12(1H,m);3.94~4.01(6H ,m);3.11~3.14(1H,m);3.02~3.05(1H,m);2.64~2.72(2H,m);1.90(3H,m);1.23~1.45(96H,m)0.86(9H,t,J=7.2Hz)

[0149] Reference Example 8 {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl(2-[{3,4,5-tris(octadecyloxy)benzoyloxy}oxy]ethyl) succinate (hereinafter referred to as "G8-suc-morT-OFF").

[0150] Step 1: Preparation of 2-hydroxyethyl 3,4,5-trioctadecyloxybenzoate 1.5 g (1.60 mmol) of 3,4,5-trioctadecyloxybenzoic acid, 370 mg (1.90 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 240 mg (1.90 mmol) of 4-(N,N-dimethylamino)pyridine were added to 8.1 mL of chloroform, followed by 120 mg (1.90 mmol) of ethylene glycol, and the mixture was stirred at room temperature for 3 hours. After the reaction was completed, 1 M aqueous sodium dihydrogen phosphate was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give 2-hydroxyethyl 3,4,5-trioctadecyloxybenzoate (882 mg; 56%).

[0151] 1 H-NMR(CDCl3):δ7.26(2H,s);4.45~4.47(2H,m);3.95~4.03(8H,m);1.25~1.52(96H,m);0.88(9H,t,J=7.2Hz)

[0152] Step 2: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl (2-[{3,4,5-tris(octadecyloxy)benzoyloxy}oxy]ethyl) succinate (hereinafter referred to as "G8-suc-morT-ON") 4-oxo-4-(2-[{3,4,5-tris(octadecyloxy)benzoyl}oxy]ethoxy)butanoic acid (hereinafter referred to as "G8-suc") was obtained in a manner similar to that of Step 1 of Reference Example 2, and then G8-suc-morT-ON was obtained in a manner similar to that of Step 2 of Reference Example 2.

[0153] 1 H-NMR(CDCl3):δ7.87(1H,bs);7.12~7.43(17H,m);6.97(1H,s);6.07~6.10(1H,m);4.33~4.46(5H,m);3.91~4.07(8H,m );3.31~3.34(1H,m);3.07~3.10(1H,m);2.56~2.60(4H,m);1.68~1.80(5H,m);1.23~1.50(96H,m);0.86(9H,t,J=7.2Hz)

[0154] Step 3: Preparation of G8-suc-morT-OFF G8-suc-morT-OFF was obtained in the same manner as in step 3 of Reference Example 2.

[0155] 1 H-NMR(CDCl3):δ7.96(1H,bs);7,23(3H,m);5.67~5.69(1H,m);4.40~4.47(5H,m);3.94~4.11(8H,m);3.09 ~3.12(1H,m);2.89~2.92(1H,m);2.53~2.65(6H,m)1.90(3H,s);1.23~1.45(96H,m);0.86(9H,t,J=6.8Hz)

[0156] Reference Example 9 {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl 4-(dioctadecylamino)-4-oxobutanoate (hereinafter referred to as "G9-suc-morT-OFF") Step 1: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl 4-(dioctadecylamino)-4-oxobutanoate (hereinafter referred to as "G9-suc-morT-ON") Using N-octadecyloctadecane-1-amine as a starting material, 4-(dioctadecylamino)-4-oxobutanoic acid (hereinafter referred to as "G9-suc") was produced in the same manner as in Step 1 of Reference Example 2. Then, G9-suc-morT-ON was produced in the same manner as in Step 2 of Reference Example 2.

[0157] 1 H-NMR(CDCl3):δ7.88(1H,bs);7.17~7.43(15H,m);6.98(1H,s);6.06~6.09(1H,m);4.31~4.35(1H,m);4.01 ~4.03(2H,m);3.08~3.34(8H,m);2.52~2.64(4H,m);1.82(3H,s);1.23~1.52(64H,m);0.85(6H,t,J=6.8Hz)

[0158] Step 2: Preparation of G9-suc-morT-OFF It was produced in the same manner as in step 3 of Reference Example 2.

[0159] 1 H-NMR(CDCl3):δ8.14(1H,bs);7.27(1H,s);5.68~5.72(1H,m):4.12~4.20(2H,m);3.98~4.01(1H,m);3.10 ~3.29(5H,m);2.94~2.97(1H,m);2.60~2.70(6H,m);1.95(3H,s);1.25~1.49(64H,m);0.88(6H,t,J=7.2Hz)

[0160] Reference Example 10: {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl 4-[{1-(octadecylamino)-1-oxo-3-phenylpropan-2-yl}amino]-4-oxobutanoate (hereinafter referred to as "G10-suc-morT-OFF"). Step 1: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl 4-[{1-(octadecylamino)-1-oxo-3-phenylpropan-2-yl}amino]-4-oxobutanoate (hereinafter referred to as "G10-suc-morT-ON") To 500 mg (1.88 mmol) of 2-tert-butoxycarbonylamino-3-phenyl-propanoic acid, 9.4 mL of tetrahydrofuran was added, followed by 652 μL (3.77 mmol) of N-ethyl-N-isopropyl-propan-2-amine, 46 mg (0.38 mmol) of 4-(N,N-dimethylamino)pyridine, 505 mg (2.64 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, and octadecane-1-amine. The mixture was stirred at room temperature for 5 hours. After completion of the reaction, 1 M aqueous sodium dihydrogen phosphate solution was added to the reaction mixture, extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give tert-butoxycarbonylamino-N-octadecyl-3-phenyl-propanamide (779 mg, 80%).

[0161] 1 H-NMR (CDCl3): δ7.18~7.28(5H,m);5.57(1H,bs);5.06(1H,bs);4.20~4.26(1H ,m);2.95~3.12(4H,m);1.40(9H,s);1.14~1.28(32H,m);0.86(3H,t,J=6.8Hz)

[0162] To 779 mg (1.51 mmol) of tert-butoxycarbonylamino-N-octadecyl-3-phenyl-propanamide, 15 mL of dichloromethane was added, followed by 1.74 mL (22.61 mmol) of trifluoroacetic acid, and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, the solvent was distilled off to obtain 2-amino-N-octadecyl-3-phenyl-propanamide (620 mg). The crude product was subjected to a reaction similar to that in Step 1 of Reference Example 2 to produce G10-suc. The crude product was then subjected to a reaction similar to that in Step 2 of Reference Example 2 to produce G10-suc-morT-ON.

[0163] 1H-NMR(CDCl3): δ8.08(1H,bs);7.16~7.32(20H,m);6.98(1H,s);6.29~6. 31(1H,m);6.10~6.12(1H,m);5.56~5.59(1H,m);4.51~4.57(1H,m);4.35~ 4.37(1H,m);4.02(2H,d,J=5.6Hz);3.73~3.77(1H,m);2.92~3.33(6H,m); 2.39~2.67(4H,m);1.84(3H,s);1.21~1.45(32H,m);0.88(3H,t,J=7.6Hz)

[0164] Process 2 Manufacturing of G10-suc-morT-OFF The target product was obtained in the same manner as in step 3 of Reference Example 2.

[0165] 1 H-NMR(CDCl3): δ8.33(1H,bs);7.16~7.32(6H,m);6.38~6.40(1H,m);5.67~5.71(2H,m);4.54~4.58(1H,m);4.08~4.16( 3H,m);3.94~4.01(1H,m);2.91~3.17(5H,m);2.46~2.79(5H,m);1.94(3H,s);1.13~1.36(32H,m);0.88(3H,t,J=7.6Hz)

[0166] The chemical structural formulas of the compounds described in Reference Examples 1 to 10 above are shown in Table 3 below.

[0167] [Table 3]

[0168] Example 1: Preparation of H-phosphonate monomers

[0169] Example 1-1: Triethylammonium [(2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl]methylphosphonate (H-phosphonate-PMO[A Bz]―ON (triethylamine salt)) and diazabicycloundecenium [(2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl]methylphosphonate (H-phosphonate-PMO[A Bz ]-ON (DBU salt) [ka] 600 mg (1.06 mmol) of N-(9-((2R,6S)-6-(hydroxymethyl)-4-tritylmorpholin-2-yl)-9H-purin-6-yl)benzamide and 1.3 mL (10.06 mmol) of N-ethylmorpholine were dissolved in 10 mL of dichloromethane. 481 μL (2.51 mmol) of diphenyl phosphite was added to this solution and stirred at room temperature for 2 hours. After 2 hours, 100 mL of 1 M triethylammonium bicarbonate solution was added and stirred at room temperature. After 1 hour, the organic layer was separated, dried over sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel chromatography to obtain H-phosphonate-PMO[A Bz ]―ON (triethylamine salt) was obtained (720 mg, 94%). ESI-MS(+):660.2(LC / MS conditions 1) 1 H-NMR(CDCl3):9.02(brs,1H);8.80(s,1H);8.01(s,2H);7.99(d,J=1.4Hz,1H);7.57~7.62(m,2H);7.49~ 7.53(m,7H);7.30(d,J=7.3,7.3Hz,6H);7.19(d,J=7.3,7.3Hz,3H);6.40(dd,J=9.6,2.3Hz,1H);4.43~4.4 9(m,1H);3.81~3.95(m,2H);3.50(ddd,J=11.0,2.3,2.3Hz,1H);3.27(ddd,J=11.9,2.3,2.3Hz,1H);2.95( q,J=7.3Hz,7.2H);1.81(dd,J=10.0,10.0Hz,1H);1.61(dd,J=11.0,11.0Hz,1H);1.23(t,J=7.3Hz,10.8H) The resulting H-phosphonate-PMO[A Bz500 mg of ]-ON (triethylamine salt) was dissolved in 5 mL of chloroform, and 20 mL of 0.2 M diazabicycloundecene bicarbonate solution was added and stirred at room temperature for 10 minutes. After 10 minutes, the organic layer was separated, dried over sodium sulfate, and the solvent was evaporated to give H-phosphonate-PMO[A Bz ]-ON (DBU salt) 490 mg was obtained. 1 H-NMR(CDCl3):9.01(brs,1H);8.80(s,1H);8.61~8.63(m,1H);8.04(s,1H);8.00(d,J=7.3Hz,2H);7.70(dddd,J=7.5,7.5 ,1.8,1.8Hz,1H);7.58~7.61(m,2H);7.49~7.53(m,6H);7.27~7.32(m,6H);7.18(d,J=7.3Hz,3H);6.40(dd,J=9.6,2.3Hz,1 H);4.44~4.49(m,1H);3.82~3.97(m,2H);3.49(ddd,J=11.0,2.3,2.3Hz,1H);3.34~3.43(m,7H);3.28(ddd,J=11.9,2.3,2. 3Hz,1H);2.80(d,J=3.2Hz,2H);1.93(dddd,J=6.0,6.0,6.0,6.0Hz,2H);1.79(dd,J=10.5,10.5Hz,1H);1.59~1.70(m,10H)

[0170] Example 1-2: Triethylammonium [(2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methylphosphonate (H-phosphonate-PMO[C Bz ]-ON), triethylammonium [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methylphosphonate (H-phosphonate-PMO[T]-ON) were prepared.

[0171] Example 2: Preparation of H-phosphonate oligomers

[0172] Example 2-1: H-phosphonate-PMO[C Bz -A Bz Production of ]-ON [ka] [ka] HO-PMO[C Bz -A Bz ]―ON is AcO-PMO[C Bz ]-ON by a known method. Bz -A Bz 3.9 g (3.7 mmol) of ]-ON was suspended in 50 mL of tetrahydrofuran. 148 μL (0.739 mmol) of 5 M sodium methoxide / methanol solution was added to this solution 10 times every 5 minutes, followed by 740 μL (3.70 mmol) of 5 M sodium methoxide / methanol solution (total: 11.09 mmol). Five minutes after the final addition, acetic acid was added and the mixture was washed with saturated brine and saturated sodium bicarbonate solution. The organic layer was dried over sodium sulfate and the solvent was evaporated. The resulting residue was dissolved in dichloromethane and reprecipitated with diisopropyl ether to obtain HO-PMO[C Bz -A Bz ]-ON was obtained (3.67g, 96%). Bz -A Bz 1.5 g (1.5 mmol) of ]-ON and 1.9 mL (15 mmol) of N-ethylmorpholine were dissolved in 15 mL of dichloromethane. 1.1 mL (5.9 mmol) of diphenyl phosphite was added to this solution and stirred overnight at 40°C. After stirring overnight, 100 mL of 1 M triethylammonium bicarbonate solution was added and stirred at room temperature. After 1 hour, the organic layer was separated, dried over sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel chromatography to obtain H-phosphonate-PMO[C Bz -A Bz ]―ON (triethylamine salt) was obtained (1.65 g, 95%). ESI-MS(+):1079.4(LC / MS conditions 1)

[0173] Example 2-2: H-phosphonate-PMO[CBz -A Bz -TC Bz Preparation of [-T]-ON [ka] AcO-PMO[C Bz -A Bz -TC Bz -T]―ON is AcO-PMO[C Bz ]-ON by a known method. Bz -A Bz -TC Bz 4 g (1.88 mmol) of [-T]-ON was suspended in 38 mL of tetrahydrofuran. 450 μL (2.26 mmol) of 5 M sodium methoxide / methanol solution was added to this solution six times every 30 minutes (total 13.56 mmol). 30 minutes after the final addition, acetic acid was added and washed with saturated brine. The organic layer was dried over sodium sulfate and the solvent was evaporated. The resulting residue was dissolved in dichloromethane and reprecipitated with diisopropyl ether to obtain HO-PMO[C Bz -A Bz -TC Bz -T]-ON was obtained (3.92 g, 99%). HO-PMO[C Bz -A Bz -TC Bz 6.2 g (3.0 mmol) of [-T]-ON and 7.4 mL (60 mmol) of N-ethylmorpholine were dissolved in 30 mL of dichloromethane. 4.6 mL (24 mmol) of diphenyl phosphite was added to this solution and stirred overnight at 40°C. After stirring overnight, 300 mL of 1 M triethylammonium bicarbonate solution was added and stirred at room temperature. After 1 hour, the organic layer was separated, dried over sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel chromatography to obtain H-phosphonate-PMO[C Bz -A Bz -TC Bz -T]―ON (triethylamine salt) was obtained (6.56 g, 98%). 1 g of the resulting product was dissolved in 5 mL of chloroform, and 20 mL of 0.2 M diazabicycloundecene bicarbonate solution was added and stirred at room temperature for 10 minutes. After 10 minutes, the organic layer was separated, dried over sodium sulfate, and the solvent was evaporated to give H-phosphonate-PMO[C Bz -A Bz -TC Bz 960 mg of [-T]-ON (DBU salt) was obtained. ESI-MS(+):2158.7(LC / MS conditions 2)

[0174] Example 2-3: H-phosphonate-PMO[C Bz -G CE,Pac -A Bz -A Bz -G CE,Pac Production of ]-ON TBDPSO-PMO[C Bz -G CE,Pac -A Bz -A Bz -G CE,Pac ]―ON is TBDPSO-PMO[C Bz ]-ON by a known method. Bz -G CE,Pac -A Bz -A Bz -G CE,Pac 19 g (7.07 mmol) of ]-ON and 3.0 mL (53.0 mmol) of acetic acid were suspended in 140 mL of tetrahydrofuran. 14.1 g (35.4 mmol) of tetrabutylammonium fluoride trihydrate was added to this solution and stirred at room temperature. After 1 hour, water was added, and the mixture was extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to obtain HO-PMO[C Bz -G CE,Pac -A Bz -A Bz -G CE,Pac ]-ON was obtained (10.7 g, 62%). Bz -G CE,Pac -A Bz -A Bz -G CE,Pac]―ON 1 g (0.408 mmol) and N-ethylmorpholine 1.0 mL (8.15 mmol) were dissolved in dichloromethane 4.0 mL. To this solution, diphenyl phosphite 937 μL (4.89 mmol) was added and stirred at room temperature. After 1 hour, 1 M triethylammonium bicarbonate solution 40 mL was added and stirred at room temperature. After 10 minutes, the organic layer was separated, dried over sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel chromatography to obtain H-phosphonate-PMO[C Bz -G CE,Pac -A Bz -A Bz -G CE,Pac ]-ON (triethylamine salt) was obtained (870 mg, 85%). 500 mg of the resulting product was dissolved in 5 mL of chloroform, and 20 mL of 0.2 M diazabicycloundecene bicarbonate solution was added and stirred at room temperature. After 10 minutes, the organic layer was separated, dried over sodium sulfate, and the solvent was evaporated to give H-phosphonate-PMO[C Bz -G CE,Pac -A Bz -A Bz -G CE,Pac ]-ON (DBU salt) 490 mg was obtained. ESI-MS(+):2606.9(LC / MS conditions 2)

[0175] Example 2-4: H-phosphonate-PMO[G CE,Pac -G CE,Pac -TTC Bz Production of ]-ON [ka] TrocO-PMO[G CE,Pac -G CE,Pac -TTC Bz 1 g (0.40 mmol) of ]-ON was dissolved in 6.0 mL of tetrahydrofuran. To this solution, 230 μL (4.01 mmol) of acetic acid and 262 mg (4.01 mmol) of zinc powder were added and stirred at room temperature. After 1 hour, 115 μL (2.01 mmol) of acetic acid and 131 mg ( 2.01(mmol) was added and stirred for 1 hour. After 1 hour, the reaction mixture was filtered through Celite, and the filtrate was evaporated. The resulting residue was purified by reverse phase purification to obtain HO-PMO[G CE,Pac -G CE,Pac -TTC Bz ]-ON was obtained (591.16 mg, 64%). HO-PMO[G CE,Pac -G CE,Pac -TTC Bz 441 mg (0.19 mmol) of ]-ON and 241 μL (1.90 mmol) of N-ethylmorpholine were dissolved in 1.9 mL of dichloromethane. 145 μL (0.76 mmol) of diphenyl phosphite was added to this solution and stirred at 40°C for 1 hour. After 1 hour, 398 μL of triethylamine was added. The reaction solution was then added dropwise to 4.5 mL of diisopropyl ether containing 2% triethylamine. After the addition, the resulting white precipitate was suction filtered and dried under reduced pressure. The resulting white powder was purified by reverse-phase purification to give H-phosphonate-PMO[G CE,Pac -G CE,Pac -TTC Bz ]-ON was obtained (211 mg, 45%). ESI-MS(+):2382.8(M+H)(LC / MS conditions 3)

[0176] Example 2-5: The following H-phosphonates were prepared in a similar manner. [Table 4]

[0177] Example 3: PMO Oligomer Preparation

[0178] Example 3-1: G1-suc-PMO[C Bz -A Bz Production of ]-ON [ka] G1-suc-PMO[C Bz ]-OFF 17 mg (0.025 mmol) and H-phosphonate-PMO[ABz 29 mg (0.0375 mmol) of ]-ON (triethylamine salt) was dissolved in 500 μL of 20% pyridine / dichloromethane. 12 μL (0.0563 mmol) of diphenyl chlorophosphate was added to this solution and stirred at room temperature. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 11.4 min, target product Rt: 17.2 min, conversion yield 92.1%, ODS condition 1). ESI-MS(+):1367.8(M+H)(LC / MS conditions 1)

[0179] Example 3-2: G1-suc-PMO[C Bz -A Bz Production of ]-ON [ka] 2 mL of solution a shown below, pumped at a flow rate of 0.25 mL / min, and 2 mL of solution b shown below, pumped at a flow rate of 0.25 mL / min, were mixed and reacted in a 5 mL tube reactor at 60 °C for 10 minutes. 100 μL of the resulting solution was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After the reaction was completed, 10% aqueous sodium thiosulfate solution was added to the reaction solution, and the organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 14.44 min, target product Rt: 17.28 min, conversion yield 95.6%, ODS condition 1). Solution a: 82 μL (0.394 mmol) of diphenyl chlorophosphate was dissolved in 2.5 mL of anhydrous dichloromethane. Solution b: H-phosphonate-PMO[A Bz ]―ON 200 mg (0.263 mmol) and G1-suc-PMO[C Bz119 mg (0.175 mmol) of ]-OFF was dissolved in 2.5 mL of a 40% anhydrous pyridine / 60% anhydrous dichloromethane solution.

[0180] Example 3-3: AcO-PMO[C Bz -C Bz Production of ]-ON [ka] AcO-PMO[C Bz ]-OFF 9 mg (0.025 mmol) and H-phosphonate-PMO[C Bz 29 mg (0.0375 mmol) of ]-ON (triethylamine salt) was dissolved in 500 μL of 20% pyridine / dichloromethane. 12 μL (0.0563 mmol) of diphenyl chlorophosphate was added to this solution and stirred at room temperature. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 5.3 min, target product Rt: 11.7, 11.8 min, conversion yield 99.7%, ODS condition 1). ESI-MS(+):1056.2(M+Na)(LC / MS conditions 1)

[0181] Example 3-4: TBSO-PMO[C Bz -C Bz Production of ]-ON [ka] TBSO-PMO[C Bz ]-OFF 11 mg (0.025 mmol) and H-phosphonate-PMO[C Bz29 mg (0.0375 mmol) of ]-ON (triethylamine salt) was dissolved in 500 μL of 20% pyridine / dichloromethane. 12 μL (0.0563 mmol) of diphenyl chlorophosphate was added to this solution and stirred at room temperature. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 10.7 min, target product Rt: 14.9, 15.0 min, conversion yield 97.7%, ODS condition 1). ESI-MS(+):1106.4(M+H)(LC / MS conditions 1)

[0182] Example 3-5: G1-suc-PMO[TG CE,Pac -A Bz Production of ]-ON [ka] G1-suc-PMO[TG CE,Pac ]-OFF 17 mg (0.0123 mmol) and H-phosphonate-PMO[A Bz 14 mg (0.0185 mmol) of ]-ON (triethylamine salt) and 20 μL (0.247 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 25 μL (0.123 mmol) of diphenyl chlorophosphate was added to this solution and stirred at 40 °C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 12.9 min, target product Rt: 18.4, 19.1 min, conversion yield 89.5%, ODS condition 2). ESI-MS(+):1819.9(LC / MS conditions 2)

[0183] Examples 3-6: G1-suc-PMO[TG CE,Pac -G CE,Pac -A Bz Production of ]-ON [ka] G1-suc-PMO[TG CE,Pac -G CE,Pac ]-OFF 21 mg (0.0125 mmol) and H-phosphonate-PMO[A Bz 14 mg (0.0188 mmol) of ]-ON (triethylamine salt) and 20 μL (0.250 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 25 μL (0.125 mmol) of diphenyl chlorophosphate was added to this solution and stirred at 40 °C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 13.6 min, target product Rt: 19.5 min, conversion yield 91.6%, ODS condition 2). ESI-MS(+):2362.0(LC / MS conditions 2)

[0184] Example 3-7: G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -A Bz Production of ]-ON G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz ]-OFF 67 mg (0.0251 mmol) and H-phosphonate-PMO[A Bz29 mg (0.0377 mmol) of ]-ON (triethylamine salt) and 10 μL (0.126 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 12 μL (0.0566 mmol) of diphenyl chlorophosphate was added to this solution and stirred at 40 °C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 14.4 min, target product Rt: 19.0, 19.2 min, conversion yield 97.9%, ODS condition 2). ESI-MS(+):3347.4(LC / MS conditions 2)

[0185] Example 3-8: G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz -A Bz ]-ON (using diphenyl chlorophosphate as a condensation agent) G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz ]-OFF 28 mg (0.00631 mmol) and H-phosphonate-PMO[A Bz9.5 mg (0.0126 mmol) of ]-ON (triethylamine salt) and 10 μL (0.126 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 13 μL (0.0631 mmol) of diphenyl chlorophosphate was added to this solution and stirred at 40 °C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 16.1 min, target product Rt: 19.5 min, conversion yield 94.4%, ODS condition 2). ESI-MS(+):5117.9 (LC / MS conditions 2)

[0186] Example 3-9: G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz -A Bz Preparation of ]-ON (using bromotripyrrolidinophosphonium hexafluorophosphate as a condensation agent) G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz ]-OFF 28 mg (0.00631 mmol) and H-phosphonate-PMO[A Bz9.5 mg (0.0126 mmol) of ]-ON (triethylamine salt) and 10 μL (0.126 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 29 mg (0.0631 mmol) of bromotripyrrolidinophosphonium hexafluorophosphate was added to this solution and stirred at 40 °C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 16.1 min, target product Rt: 19.5 min, conversion yield 94.0%, ODS condition 2).

[0187] Example 3-10: G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz -A Bz Preparation of ]-ON (using diphenylbis(2,6-dimethylphenyl) chlorophosphate as a condensation agent) G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz ]-OFF 28 mg (0.00631 mmol) and H-phosphonate-PMO[A Bz9.5 mg (0.0126 mmol) of ]-ON (triethylamine salt) and 10 μL (0.126 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 21 mg (0.0631 mmol) of diphenylbis(2,6-dimethylphenyl)chlorophosphate was added to this solution and stirred at 40 °C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 16.1 min, target product Rt: 19.5 min, conversion yield 97.0%, ODS condition 2).

[0188] Example 3-11: G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz -A Bz Preparation of ]-ON (using bis(2-oxo-3-oxazolidinyl)phosphinic chloride as a condensation agent) G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz ]-OFF 28 mg (0.00631 mmol) and H-phosphonate-PMO[A Bz9.5 mg (0.0126 mmol) of ]-ON (triethylamine salt) and 10 μL (0.126 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 16 mg (0.0631 mmol) of bis(2-oxo-3-oxazolidinyl)phosphinic chloride was added to this solution and stirred at 40 °C. After 10 min, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 min. After completion of the reaction, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 16.1 min, target product Rt: 19.5 min, conversion yield 95.9%, ODS condition 2).

[0189] Example 3-12: G1-suc-PMO[TA Bz ]-ON (using piperidine instead of dimethylamine in the oxidizing solution) [ka] G1-suc-PMO[T]-OFF 15 mg (0.025 mmol) and H-phosphonate-PMO[A Bz 29 mg (0.0375 mmol) of ]-ON (triethylamine salt) and 20 μL (0.253 mmol) of N-methylimidazole were dissolved in 500 μL of dichloromethane. 26 μL (0.127 mmol) of diphenyl chlorophosphate was added to this solution and stirred at room temperature. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M piperidine / THF solution) and stirred at room temperature for 10 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 12.0 min, target product Rt: 19.6, 20.1 min, conversion yield 98.9%, ODS condition 2). ESI-MS(+):1317.7(LC / MS conditions 2)

[0190] Example 3-13: The following oligomers were prepared in a similar manner. [Table 5] Example 4: Preparation of PMO Oligomers

[0191] Example 4-1: G1-suc-PMO[A Bz -C Bz Preparation of [-TT]-ON [ka] G1-suc-PMO[A Bz -C Bz 28 mg (0.025 mmol) of ]-OFF and 37 mg (0.0375 mmol) of H-phosphonate-PMO[TT]-ON (triethylamine salt) were dissolved in 500 μL of 20% pyridine / dichloromethane. 12 μL (0.0563 mmol) of diphenyl chlorophosphate was added to this solution and stirred at room temperature. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 13.8 min, target product Rt: 15.3 min, conversion yield 97.2%, ODS condition 1). ESI-MS(+):2026.9(LC / MS conditions 2)

[0192] Example 4-2: G1-suc-PMO[A Bz -C Bz Preparation of [-TT]-ON [ka] 2 mL of solution a shown below, pumped at a flow rate of 0.2 mL / min, and 2 mL of solution b shown below, pumped at a flow rate of 0.2 mL / min, were mixed and reacted in a 5 mL tube reactor at 60 °C for 12.5 minutes. 100 μL of the resulting solution was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After the reaction was completed, 10% aqueous sodium thiosulfate solution was added to the reaction solution, and the organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 13.79 min, target product Rt: 15.27 min, conversion yield 95.0%, ODS condition 1). Solution a: 82 μL (0.394 mmol) of diphenyl chlorophosphate was dissolved in 2.5 mL of anhydrous dichloromethane. Solution b: H-phosphonate-PMO[TT]-ON 257 mg (0.263 mmol) and G1-suc-PMO[A Bz -C Bz 197 mg (0.175 mmol) of ]-OFF was dissolved in 2.5 mL of a 40% anhydrous pyridine / 60% anhydrous dichloromethane solution.

[0193] Example 4-3: G1-suc-PMO[TTA Bz Preparation of [-TT]-ON [ka] G1-suc-PMO[TTA Bz34 mg (0.025 mmol) of ]-OFF and 37 mg (0.0375 mmol) of H-phosphonate-PMO[TT]-ON (triethylamine salt) were dissolved in 500 μL of 20% pyridine / dichloromethane. 12 μL (0.0563 mmol) of diphenyl chlorophosphate was added to this solution and stirred at room temperature. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 12.4 min, target product Rt: 14.2 min, conversion yield 88.9%, ODS condition 1). ESI-MS(+):2268.0(LC / MS conditions 2)

[0194] Example 4-4: G1-suc-PMO[TTA Bz Preparation of [-TT]-ON [ka] 2 mL of solution a shown below, pumped at a flow rate of 0.2 mL / min, and 2 mL of solution b shown below, pumped at a flow rate of 0.2 mL / min, were mixed and reacted in a 5 mL tube reactor at 60 °C for 12.5 minutes. 100 μL of the resulting solution was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After the reaction was completed, 10% aqueous sodium thiosulfate solution was added to the reaction solution, and the organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 12.42 min, target product Rt: 14.24 min, conversion yield 94.1%, ODS condition 1). Solution a: 82 μL (0.394 mmol) of diphenyl chlorophosphate was dissolved in 2.5 mL of anhydrous dichloromethane. Solution b: H-phosphonate-PMO[TT]-ON 257 mg (0.263 mmol) and G1-suc-PMO[TTA Bz239 mg (0.175 mmol) of ]-OFF was dissolved in 2.5 mL of a 40% anhydrous pyridine / 60% anhydrous dichloromethane solution.

[0195] Example 4-5: G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -C Bz -A Bz -TC Bz Preparation of [-T]-ON (using triethylamine salt of H-phosphonate) G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz ]-OFF 33 mg (0.0123 mmol) and H-phosphonate-PMO[C Bz -A Bz -TC Bz 40 mg (0.0185 mmol) of [-T]-ON (triethylamine salt) and 20 μL (0.247 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 26 μL (0.124 mmol) of diphenyl chlorophosphate was added to this solution and stirred at 40 °C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 14.4 min, target product Rt: 18.6 min, conversion yield 84.7%, target product purity 23.4%, ODS condition 2). ESI-MS(+):4845.8 (LC / MS conditions 2)

[0196] Examples 4-6: G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -C Bz -A Bz -TC Bz Preparation of [-T]-ON (using DBU salt of H-phosphonate) G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz ]-OFF 33 mg (0.0123 mmol) and H-phosphonate-PMO[C Bz -A Bz -TC Bz 40 mg (0.0185 mmol) of [-T]-ON (DBU salt) and 20 μL (0.247 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 26 μL (0.124 mmol) of diphenyl chlorophosphate was added to this solution and stirred at 40 °C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 14.4 min, target product Rt: 18.6 min, conversion yield 95.8%, target product purity 53.1%, ODS condition 2).

[0197] Examples 4-7: G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz -C Bz -G CE,Pac -A Bz -A Bz -G CE,Pac Production of ]-ON G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz ]-OFF 28 mg (0.00631 mmol) and H-phosphonate-PMO[C Bz -G CE,Pac -A Bz -A Bz -G CE,Pac32 mg (0.0126 mmol) of ]-ON (DBU salt) and 16 μL (0.126 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 13 μL (0.0631 mmol) of diphenyl chlorophosphate was added to this solution and stirred at 40 °C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate solution was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 16.1 min, target product Rt: 19.8 min, conversion yield 91.5%, ODS condition 2). ESI-MS(+):7064.4(LC / MS conditions 2)

[0198] Examples 4-8: The following oligomers were prepared in a similar manner. [Table 6]

[0199] Example 5: G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -G CE,Pac -G CE,Pac -TTC Bz -TG CE,Pac -A Bz -A Bz -G CE,Pac -G CE,Pac -TG CE,Pac Preparation of [-TT]-ON [ka] TIFF0007825613000097.tif93122 TIFF0007825613000098.tif145154G1-suc-PMO[C Bz -C Bz -TC Bz -C Bz -GCE,Pac -G CE,Pac -TTC Bz ]-OFF 28 mg (0.00631 mmol) and H-phosphonate-PMO[TG CE,Pac -A Bz -A Bz -G CE,Pac -G CE,Pac -TG CE,Pac 43 mg (0.00947 mmol) of [-TT]-ON (DBU salt) and 16 μL (0.126 mmol) of N-methylimidazole were dissolved in 250 μL of dichloromethane. 13 μL (0.0631 mmol) of diphenyl chlorophosphate was added to this solution and stirred at 40 °C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidizing solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After the reaction was complete, 10% aqueous sodium thiosulfate was added to the reaction mixture. The organic and aqueous layers were separated. The organic layer was diluted 20-fold with 80% acetonitrile and analyzed by HPLC (raw material Rt: 15.2 min, target product Rt: 17.6 min, conversion yield 91.0%, ODS condition 2). To 10 μL of the reaction solution, 190 μL of a 3:1 mixture of aqueous ammonia and ethanol was added, followed by stirring for 18 hours at 55° C. The reaction solution was filtered, and the filtrate was diluted 50-fold with 20% aqueous acetonitrile and subjected to MS measurement, confirming the production of the target product. ESI-MS(+):6848.4(LC / MS conditions 1)

Claims

1. Formula [A-1]: 【Chemistry 1】 [In the formula, B Ｐ are the same or different, and each is an optionally protected nucleobase; n is an integer from 1 to 50; W may be the same or different and is an oxygen atom or a sulfur atom; and X's may be the same or different and each represent a hydroxyl group substituted with a group that can be removed under neutral conditions, 1,1,3,3-tetra(C １－６ alkyl) guanidyl, C １－６ Alkoxy, di(C １－６ alkyl)amino, mono(amino-C substituted with a group that can be removed under basic conditions １－６ alkyl)amino, di(amino-C substituted with a group that can be removed under basic conditions １－６ alkyl)amino, or general formula [2]: 【Chemistry 2】 (In the formula, * indicates the bonding position with P; a is an integer from 0 to 2; E is CH ２ , CH-A １ or N-A ２ and A １ is C １－６ Alkyl, mono(C １－６ alkyl)amino-C １－６ Alkyl, di(C １－６ alkyl)amino-C １－６ Alkyl, tri(C １－６ Alkyl)ammonio-C １－６ alkyl, amino substituted with a group removable under basic conditions, mono (C １－６ alkyl)amino, di(C １－６ alkyl)amino, tri(C １－６ alkyl)ammonio, amino substituted with amidino substituted with a group removable under basic conditions, or a compound represented by the general formula [3]: 【Transformation 3】 (In the formula, * indicates the bonding position with E; b is an integer from 0 to 2; c is 0 or 1; R １１ is C １－６ is alkyl; and M is CH ２ , an oxygen atom, a sulfur atom, or N-(a group removable under basic conditions); and A ２ is C １－６ Alkyl, mono(C １－６ alkyl)amino-C １－６ Alkyl, di(C １－６ alkyl)amino-C １－６ Alkyl, tri(C １－６ Alkyl)ammonio-C １－６ alkyl, a group removable under basic conditions, aryl, or heteroaryl) is a substituent represented by G is, (1) silicon-substituted groups, (2) optionally substituted C １－１８ Alkyl-carbonyl, optionally substituted C １－１８ alkoxy-carbonyl, long chain alkyl-carbonyl, or long chain alkoxy-carbonyl; (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; or (4) General formula [7]: 【Chemistry 4】 (In the formula, * indicates the binding position to T; Z is (1) (Soluble polymer soluble in organic solvent)-oxy, (2) (Soluble polymer soluble in organic solvent)-amino, (3) long-chain alkyloxy, benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or benzyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; (4) a solid support, or (5) The following general formulas [8A] to [8N]: 【Transformation 5】 (In the formula, * indicates the bonding position with L; j is an integer from 0 to 4; k is an integer from 0 to 5; R ８ａ is a hydrogen atom or C １－６ is alkyl; R ８ｂ are the same or different and are long chain alkyl; R ８ｃ are the same or different and represent the following general formulae [9A] to [9E]: 【Transformation 6】 (In the formula, * is the bond position; and R ９ is a long chain alkyl and / or long chain alkenyl) selected from the substituents represented by R ８ｄ are the same or different and each represent a hydrogen atom, a halogen atom, a long-chain alkyl optionally substituted with 1 to 13 halogen atoms, or a long-chain alkyloxy optionally substituted with 1 to 13 halogen atoms; R ８ｅ teeth (1) long-chain alkyl, (2) a long chain alkyl-carbonyl, or (3) benzoyl substituted with 1 to 5 long chain alkyloxy and / or long chain alkenyloxy; and R ８ｆ teeth, (1) long-chain alkyl, (2) a long chain alkyl-carbonyl, or (3) Long-chain alkenyl-carbonyl selected from the substituents represented by and L is a group represented by the general formula [10]: 【Transformation 7】 (In the formula, * is the bonding position with Z; ** is the bonding position with T; and L １ is an optionally substituted C ２－１０ Alkylene or optionally substituted C ６－１０ arylene), or general formula [10-1]: 【Transformation 8】 (In the formula, * is the bonding position with Z; ** is the bonding position with T; and L １ is an optionally substituted C １－１０ Alkylene or optionally substituted C ６－１０ arylene) is a substituent represented by is a substituent represented by the formula: T is a single bond or a group represented by the general formula [11]: 【Chemistry 9】 (In the formula, X and W are as defined above; * indicates the bond position to O; ** is the bond position to G; and q is an integer from 0 to 10. and when G is a silicon substituent, T is a single bond. and a compound of formula [B-1]: 【Chemistry 10】 [In the formula, B Ｐ , W, and X are as defined above, and p is an integer from 1 to 50; L' is OH, -O － N ＋ H (aliphatic amine), —O － N ＋ H (cyclic amine), or —O － N ＋ H (aromatic amine); Q １ is a group that can be removed under acidic conditions. and a compound of (1) treating with at least one condensing agent selected from the group consisting of phosphorus reagent 1, phosphorus reagent 2, and onium reagent in the presence of a base; and (2) Treating with an oxidizing agent and an organic amine, Formula [C-1]: 【Chemistry 11】 [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above. A method for obtaining a compound of the formula The phosphorus reagent 1 is a compound represented by the following formula: 【Chemistry 12】 [In the formula, W is an oxygen atom or a sulfur atom; R １ and R ２ are the same or different and are selected from the group consisting of an optionally substituted aromatic ring -O- and an optionally substituted heterocycle. is shown by The phosphorus reagent 2 is a compound represented by the following formula: 【Chemistry 13】 [In the formula, R １ , R ２ , R ３ , and R ４ are the same or different and are halogen, C １－６ Alkylamino, di(C １－６ alkyl)amino, optionally substituted cyclic C １－６ selected from the group consisting of amino, optionally substituted aromatic amino, optionally substituted aromatic ring, and optionally substituted heterocycle-O—; or R １ and R ２ together with the phosphorus atom to which they are attached form an optionally substituted ring structure, and R ３ and R ４ are the same or different and are halogen, C １－６ Alkylamino, di(C １－６ alkyl)amino, optionally substituted cyclic C １－６ amino, optionally substituted aromatic amino, optionally substituted aromatic ring, and optionally substituted heterocycle-O—. is shown by The onium reagent has the formula: 【Chemistry 14】 [In the formula, R １ , R ２ and R3 are the same or different and each represents a halogen atom, C １－６ Alkylamino, di(C １－６ alkyl)amino, optionally substituted cyclic C １－６ Amino, optionally substituted aromatic amino, optionally substituted aromatic ring, optionally substituted heterocycle-O-, and optionally substituted C １－６ alkylideneaminooxy; or R １ and R ２ together with the carbon atoms to which they are attached form a ring structure, and R ３ is a halogen, C １－６ Alkylamino, di(C １－６ alkyl)amino, optionally substituted cyclic C １－６ amino, optionally substituted aromatic amino, optionally substituted aromatic ring, and optionally substituted heterocycle-O—. The method shown in

2. Furthermore, the formula [C-1]: 【Chemistry 15】 [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above. with an acid, or an acid and a scavenger, to Formula [C-0-1]: 【Chemistry 16】 [In the formula, B Ｐ , W, X, G, T, n, and p are as defined above. compound of, and The compound of formula [C-0-1] and a compound of formula [B-0-1]: 【Chemistry 17】 [In the formula, B Ｐ , Q １ , W, and L′ are as defined above. and a compound of (1) treating with at least one condensing agent selected from the group consisting of the phosphorus reagent 1, the phosphorus reagent 2, and the onium reagent in the presence of a base, and then (2) Treating with an oxidizing agent and an organic amine, Formula [C-1-1]: [Chemistry 18] [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above.

10. The method of claim 1, comprising obtaining a compound of formula:

3. Furthermore, the formula [C-1]: 【Chemistry 19】 [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above. The compound is treated under conditions for removing a protecting group of a hydroxyl group, Formula [C-0-2]: 【Chemistry 20】 [In the formula, B Ｐ , Q １ , W, X, n, and p are as defined above. to obtain a compound of (1) Formula [P-1]: 【Chemistry 21】 wherein W is an oxygen atom or a sulfur atom, and R １ and R ２ are the same or different and each represents H, optionally substituted C １－６ alkyl, optionally substituted phenyl, and PH(=O)-OH. and a base, and subsequently (2) Treating with a hydrolysis solution, Formula [C-0-2-1]: 【Chemistry 22】 [In the formula, B Ｐ , Q １ , W, X, L′, n, and p are as defined above.

10. The method of claim 1, comprising obtaining a compound of formula:

4. Furthermore, the formula [C-0-2-1]: 【Chemistry 23】 [In the formula, B Ｐ , Q １ , W, X, L′, n, and p are as defined above. and a compound of formula [A-0-1]: 【Chemistry 24】 [In the formula, B Ｐ , G, and T are as defined above. and a compound of (1) treating with at least one condensing agent selected from the group consisting of the phosphorus reagent 1, the phosphorus reagent 2, and the onium reagent in the presence of a base, and then (2) Treating with an oxidizing agent and an organic amine, Formula [C-1-2]: 【Chemistry 25】 [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above.

4. The method of claim 3, comprising obtaining a compound of formula:

5. Furthermore, the formula [B-1-0]: 【Chemistry 26】 [In the formula, B Ｐ , Q １ , W, X, and p are as defined above. The compound (1) Formula [P-1]: 【Chemistry 27】 [Wherein W is an oxygen atom or a sulfur atom, R １ , and R ２ are the same or different and each represents H, optionally substituted C １－６ alkyl, optionally substituted phenyl, and PH(=O)-OH. and a base, and subsequently (2) Treating with a hydrolysis solution, Formula [B-1]: 【Chemistry 28】 [In the formula, B Ｐ , Q １ , W, X, L′, and p are as defined above.

10. The method of claim 1, comprising obtaining a compound of formula:

6. Furthermore, the formula [A-0]: 【Chemistry 29】 [In the formula, B Ｐ , Q １ , W, X, G, T, and n are as defined above. with an acid, or an acid and a scavenger, to Formula [A-1]: 【Transformation 30】 [In the formula, B Ｐ , W, X, G, T, and n are as defined above.

10. The method of claim 1, comprising obtaining a compound of formula:

7. G is a group having the following formula: 【Chemistry 31】 [In the formula, * represents the bonding position with T.] selected from the group consisting of: T is a single bond; The method according to any one of claims 1 to 6.

8. Formula [B-1] 【Chemistry 32】 [In the formula, B Ｐ are the same or different and are optionally protected nucleobases, Q １ is trityl or dimethoxytrityl, L' is -O － N ＋ H (CH ２ CH ３ ) ３ , or -O － (HDBU) ＋ and W is O, X's may be the same or different and each represent a hydroxyl group substituted with a group that can be removed under neutral conditions, 1,1,3,3-tetra(C １－６ alkyl) guanidyl, C １－６ Alkoxy, di(C １－６ alkyl)amino, mono(amino-C substituted with a group that can be removed under basic conditions １－６ alkyl)amino, di(amino-C substituted with a group that can be removed under basic conditions １－６ alkyl)amino, or general formula [2]: 【Transformation 33】 (In the formula, * indicates the bonding position with P; a is an integer from 0 to 2; E is CH ２ , CH-A １ or N-A ２ and A １ is C １－６ Alkyl, mono(C １－６ alkyl)amino-C １－６ Alkyl, di(C １－６ alkyl)amino-C １－６ Alkyl, tri(C １－６ Alkyl)ammonio-C １－６ alkyl, amino substituted with a group removable under basic conditions, mono (C １－６ alkyl)amino, di(C １－６ alkyl)amino, tri(C １－６ alkyl)ammonio, amino substituted with amidino substituted with a group removable under basic conditions, or a compound represented by the general formula [3]: 【Transformation 34】 (In the formula, * indicates the bonding position with E; b is an integer from 0 to 2; c is 0 or 1; R １１ is C １－６ is alkyl; and M is CH ２ , an oxygen atom, a sulfur atom, or N-(a group removable under basic conditions); and A ２ is C １－６ Alkyl, mono(C １－６ alkyl)amino-C １－６ Alkyl, di(C １－６ alkyl)amino-C １－６ Alkyl, tri(C １－６ Alkyl)ammonio-C １－６ alkyl, a group removable under basic conditions, aryl, or heteroaryl) and p is an integer from 2 to 20. A compound represented by the formula:

9. ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate, triethylammonium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate, ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl (2H)-yl)methoxy)phosphoryl)morpholin-2-yl)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)phosphoryl)-6-(6-benzamido-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate, Triethylammonium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpho 9H-purin-9-yl)morpholin-2-yl)methoxy)phosphoryl)morpholin-2-yl)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)phosphoryl)-6-(6-benzamido-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate, ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-((((2S,6R)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)-4-to ritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate, Triethylammonium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-((((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-((((2S,6R)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl )-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate, ((2S,6R)-4-((((2S,6R)-4-((((2S,6R)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(5-methyl-2,4 -dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methylphosphonate, Triethylammonium ((2S,6R)-4-((((2S,6R)-4-((((2S,6R)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(5-methyl ethyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl)methylphosphonate, ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate, triethylammonium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate, ((2S,6R)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate, triethylammonium ((2S,6R)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate, ((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate, triethylammonium ((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate, ((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)((((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate, triethylammonium ((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)(((2S,6R)-4-((dimethylamino)((((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methylphosphonate, 1,8-Diazabicyclo(5.4.0)-7-undecenium((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((((2S,6R)-4-((((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4- tritylmorpholin-2-yl)methoxy)phosphoryl)morpholin-2-yl)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)phosphoryl)-6-(6-benzamido-9H-purin-9-yl)morpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methylphosphonate, and 1,8-diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-4-(((2S,6R)-4-(((2S,6R)-6-(6-benzamidopurin-9-yl)-4-(((2S,6R)-6-(6-benzamidopurin-9-yl)-4-(((2S,6R)-4-(((2S,6R)-6-(6-(2-cyanoethoxy)-2-((2-phenoxyacetyl)amino)purin-9-yl)-4-(((2S,6R)-4- (((2S,6R)-6-(6-(2-cyanoethoxy)-2-((2-phenoxyacetyl)amino)purin-9-yl-4-(dimethylamino-(((2S,6R)-4-(dimethylamino-(((2S,6R)-6-(5-methyl-2,4-dioxo-pyrimidin-1-yl)-4-trityl-morpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-pyrimidin-1-yl)morpholin-2-yl)meth methoxy-(dimethylamino)phosphoryl-6-(5-methyl-2,4-dioxo-pyrimidin-1-yl)morpholin-2-yl)methoxy-(dimethylamino)phosphoryl)morpholin-2-ylmethoxy-(dimethylamino)phosphoryl)-6-(6-(3-cyanopropoxy-2-((2-phenoxyacetyl)amino)purin-9-yl)morpholin-2-yl)methoxy-(dimethylamino) 6-(6-(2-cyanoethoxy)-2-((2-phenoxyacetyl)amino)purin-9-yl)morpholin-2-yl)methoxy-(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-pyrimidin-1-yl)morpholin-2-yl)methoxyphosphinate A compound selected from the group consisting of:

10. Formula [B-0-1]: 【Chemistry 35】 [In the formula, B Ｐ is an optionally protected nucleobase, Q １ is trityl or dimethoxytrityl, L' is -O － (HDBU) ＋ and W is O. Compound.

11. 1,8-diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methylphosphonate, 1,8-diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-(4-benzamido-2-oxopyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methylphosphonate, 1,8-diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methylphosphonate, 1,8-diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-((2-cyanoethoxy)-2-(2-phenoxyacetamido)-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methylphosphonate, and 1,8-diazabicyclo(5.4.0)-7-undecenium ((2S,6R)-6-(2-(2-phenylacetamido)-6-((4-(pivaloyloxy)benzyl)oxy)-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methylphosphonate The compound of formula [B-0-1] according to claim 10, selected from the group consisting of:

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