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

Method for producing oligonucleic acid compound

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

[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 by contacting a substrate supported on a solid support with a solution containing a reaction reagent. The solid-phase method uses a so-called batch method in which a reaction is carried out in a reaction vessel equipped with a filter (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 a solid support is placed in a column and a solution containing a reaction reagent is passed through the column to carry out the reaction. Meanwhile, the liquid-phase method is a homogeneous reaction method in which nucleic acids are extended by reaction 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 vessel (see, for example, Patent Documents 2 and 3).

[0003] In chemical synthesis of oligonucleotide compounds, whether solid-phase, liquid-phase, batch, or pseudo-flow synthesis, nucleic acids are extended by repeatedly repeating a "deprotection" reaction, which removes the protecting groups of oxygen atoms or amino groups on the nucleic acid compound, and a "condensation" reaction, which forms a bond between the deprotected oxygen atom or nitrogen atom, which has become reactive, and the phosphorus atom. In particular, controlling the reaction efficiency and reaction rate of the "condensation" reaction, which forms a bond between the phosphorus atom and the oxygen atom or nitrogen atom, is extremely important in the production of oligonucleotide compounds, and the conditions of this condensation reaction are factors that have a significant impact on the production time of oligonucleotide compounds.

[0004] Because the solid-phase method involves 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 commonly used as the solid support, but during the reaction, it swells in the reaction solvent used, increasing its volume 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, polystyrene resin does not swell very much in polar solvents such as acetonitrile, which are commonly used in the synthesis of oligonucleic acid compounds. Therefore, using a polar solvent in the solid-phase method is not desirable from the perspective of improving the reaction efficiency and reaction rate of the condensation reaction.

[0005] On the other hand, known homogeneous reaction methods include liquid-phase methods and synthesis methods using hydrophobic-group-bonded nucleosides, pseudo-solid-phase protected nucleosides, etc. The liquid-phase method is a homogeneous reaction method in which a reaction is carried out in a solution containing both a substrate and a reaction reagent. Compared to solid-phase methods, the liquid-phase method offers higher reaction efficiency and a faster reaction rate, but requires column purification or the like to remove impurities such as reaction reagents and reaction solvents. Similar to the liquid-phase method, synthesis methods using hydrophobic-group-bonded nucleosides, pseudo-solid-phase protected nucleosides, etc., allow the reaction to occur in a homogeneous system, resulting in higher reaction efficiency and a faster reaction rate 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).

[0006] International Publication No. 1991 / 09033A1 International Publication No. 2014 / 077292A1 International Publication No. 2013 / 122236A1 Japanese Patent No. 5548852 International Publication No. 2016 / 060135A1

[0007] Acc. Chem. Res. , Vol. 24, 278-284, 1991

[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.

[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]: [In the formula, B Ｐ are the same or different and each represents an optionally protected nucleobase; 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 is the same or different and each represents an oxygen atom or a sulfur atom; and X is the same or different and each represents a hydroxyl group substituted with a group removable 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]: (Wherein, * is the bonding position to P; a is an integer of 0 to 2; E is CH ２ , CH-A １ or N-A ２ and 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 １－６alkyl)ammonio, amino substituted with amidino substituted with a group removable under basic conditions, or a compound represented by the general formula [3]: (wherein * is the bonding position with E; b is an integer of 0 to 2; c is 0 or 1; R １１ is C １－６ 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 G is a substituent represented by the formula (1), which is an alkyl, a group removable under basic conditions, an aryl, or a heteroaryl; G is (1) a silicon substituent, (2) an 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) a group represented by the general formula [7]: (wherein * is the bonding position with T; Z is: (1) (a soluble polymer that dissolves in an organic solvent)-oxy, (2) (a soluble polymer that dissolves in an organic solvent)-amino, (3) a long-chain alkyloxy, a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or a benzyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, (4) a solid phase support, or (5) a group represented by the following general formulas [8A] to [8N]: (wherein * is the bonding position to L; j is an integer of 0 to 4; k is an integer of 0 to 5; R ８ａis a hydrogen atom or C １－６ 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]: (wherein * is the bond position; and R ９ is a long chain alkyl and / or long chain alkenyl; ８ｄ are the same or different and each represent a hydrogen atom, a halogen atom, a long-chain alkyl group optionally substituted with 1 to 13 halogen atoms, or a long-chain alkyloxy group optionally substituted with 1 to 13 halogen atoms; R ８ｅ is (1) a long chain alkyl, (2) a long chain alkyl-carbonyl, or (3) a benzoyl substituted with 1 to 5 long chain alkyloxy and / or long chain alkenyloxy; and R ８ｆ is a substituent represented by the general formula

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

[11] : (wherein X and W are as defined above; * is the bonding position to **O, *O, or *N in the above general formulae [4a] to [4d]; ** is the bonding position to G; and q is an integer of 0 to 10), but when G is a silicon substituent, T is a single bond), with a compound of formula [B-1]: [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 (CH ２ CH ３ ) ３ and -O － N ＋ H (cyclic amine) is, for example, —O － (HDBU) ＋ and Q １ is a group removable under acidic conditions], and a compound of formula [C-1]: (1) treated 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) treated with an oxidizing agent and an organic amine to obtain a compound of formula [C-2]: [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above.

[0012] <2> Further, the formula [C-1]: [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 obtain a compound of formula [C-0-1]: [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above], and [In the formula, B Ｐ , Q １ , W, and L′ are as defined above] 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) with an oxidizing agent and an organic amine to obtain a compound of formula [C-1-1]: [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above.

[0013] <3> Further, the formula [C-1]: [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above] is treated under conditions for removing a hydroxyl-protecting group to obtain a compound of formula [C-0-2]: [In the formula, B Ｐ , Q １ , W, X, n, and p are as defined above], and (1) a compound of formula [P-1]: 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 (2) treating with a compound of formula [C-0-2-1], wherein the compound is selected from the group consisting of alkyl, optionally substituted phenyl, PH(=O)-OH, and a base, and subsequently with a hydrolysis solution to produce a compound of formula [C-0-2-2]: [In the formula, B Ｐ , Q １, W, X, L', n, and p are as defined above.

[0014] <4> Further, the formula [C-0-2-1]: [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]: [In the formula, B Ｐ , G, and T are as defined above] 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) with an oxidizing agent and an organic amine to obtain a compound of formula [C-1-2]: [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above.

[0015] <5> Further, the formula [B-1-0]: [In the formula, B Ｐ , Q １ , W, X, and p are as defined above] with a compound of formula [P-1]: [In the formula, W, R １ , and R ２ has the same meaning as above] and a base, and subsequently, (2) treating with a hydrolysis solution to obtain a compound of formula [B-1]: [In the formula, B Ｐ , Q １ , W, X, L', and p are as defined above.

[0016] <6> Furthermore, the formula [A-0]: [In the formula, B Ｐ , Q １ , W, X, G, T, and n are as defined above] is treated with an acid, or an acid and a scavenger, to obtain a compound of formula [A-1]: [In the formula, B Ｐ , W, X, G, T, and n are as defined above.

[0017] <7> G is the following formula: [wherein * represents the bonding position to T]; and T is a single bond.

[0018] <8> In formula [B-1], B Ｐ are the same or different and are optionally protected nucleobases, and Q １ is trityl or dimethoxytrityl, and L' is -O － N ＋ H (CH ２ CH ３ ) ３ , or -O － (HDBU) ＋ The compound of formula [B-1] according to <1>, wherein 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.

[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 -T-C 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) (2H)-trimethyl-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 -T-C 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[T-T]-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[T-T]-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[T-T-T]-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[T-T-T]-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 pyrimidin-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[T-T-T-T]-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) 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 (i.e., H-phosphonate-PMO[T-T-T-T]-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-tritylmorpholin-2-yl)methoxy)phosphoryl)morpholin-2-yl)(dimethylamino)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl),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 -T-C 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-pyrimidin-1-yl)-4-trityl-morpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-pyrimidin-1-yl)morpholin-2-yl)methoxy)phosphoryl)mo 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)purin-9-yl)morpholin-2-yl)methoxy-(dimethylamino)phosphoryl)morpholin-2 -yl)methoxy-(dimethylamino)phosphoryl)morpholin-2-yl)methoxy-(dimethylamino)phosphoryl)-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., H-phosphonate-PMO[T-G ＣＥ，Ｐａｃ -A Ｂｚ -A Ｂｚ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -T-G ＣＥ，Ｐａｃ -TT]-ON (DBU salt)).

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

[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, The compound of formula [B-0-1] according to <10> is selected from the group consisting of 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.

[0022] An oligonucleic acid compound is a compound 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 shorter production time for an oligonucleic acid compound.

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

[0024] Explanation of Terms: The "nucleobase" can include, for example, adenine, guanine, hypoxanthine, cytosine, thymine, uracil, or 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 according to the formula (I) may be protected. "Optionally protected nucleic acid base" encompasses both unprotected and protected "nucleobases," and examples include adenine, guanine, hypoxanthine, cytosine, thymine, uracil, etc., in which the amino group and / or hydroxyl group is unprotected or protected. The amino group protecting group is not particularly limited as long as it is used as a protecting group for nucleic acids, and specific examples include benzoyl, 4-methoxybenzoyl, acetyl, propionyl, butyryl, isobutyryl, phenylacetyl, phenoxyacetyl, 4-tert-butylphenoxyacetyl, 4-isopropylphenoxyacetyl, and (dimethylamino)methylene. Preferred amino group 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 are 2-cyanoethyl, 4-nitrophenethyl, and 4-(tert-butylcarboxy)benzyl. The preferred protecting group for the 6-hydroxyl group of guanine is 2-cyanoethyl. In one embodiment, examples of protected nucleobases include those shown below. [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 whose amino group is protected with 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. "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" described 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 "5- to 6-membered saturated cyclic amino" include a 5- to 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, specifically 1-pyrrolidinyl, 1-imidazolidinyl, piperidino, 1-piperazinyl, 1-tetrahydropyrimidinyl, 4-morpholino, 4-thiomorpholino, 1-homopiperazinyl, and oxazolidin-3-yl. １－６ "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. １－６ The 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. １－６ Alkoxy-C １－６ "C" in "Alkyl" １－６ The "alkoxy" portion is the same as the "C１－６ The same as "alkoxy" can be mentioned. 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" moiety is １－６ The same as "C alkyl" can be mentioned. 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 an ethylene group, a propylene group, an isopropylene group, a butylene group, a pentylene group, and a hexylene group. Such an "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. ６－１０ "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. １－６ alkyl)guanidyl", "C１－６ 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" moiety is １－６Examples of "groups removable under acidic conditions" include trityl, monomethoxytrityl, tert-butyldimethylsilyl, and dimethoxytrityl. Examples of "groups removable under basic conditions" include trifluoroacetyl. Examples of "groups 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 "silicon substituents" include triphenylsilyl, diisopropylphenylsilyl, tert-butyldimethylsilyl, and tert-butyldiphenylsilyl. Examples of "aryl" include 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 a "soluble polymer that dissolves in an organic solvent" 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 with a molecular weight of 100 to 40,000 and having a substituent (SUNBRIGHT (registered trademark) series).

[0025] <Elongation 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]: [In the formula, B Ｐ , Q １ , W, X, G, T, n, L′, and p are as defined above], and the method may comprise: step (1): condensation; and step (2): oxidative amination.

[0026] One aspect of the present invention is a compound represented by formula [A-1]: [In the formula, B Ｐ , W, X, G, T, n, and p are as defined above] and a compound of formula [B-1]: [In the formula, B Ｐ , Q １ , W, X, L′, and p are as defined above], and a compound of formula [C-1]: [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above.

[0027] <Step (1): Condensation>

[0028] <Condensing Agent in 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 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 even more preferably diphenyl chlorophosphate, bis(2,6-dimethylphenyl) chlorophosphate, or PyBrop.

[0030] In one embodiment of the present invention, the phosphorus reagent 1 is represented by the formula [P-2]: wherein W is an oxygen atom or a sulfur atom; １ and R ２are the same or different and selected from the group consisting of an optionally substituted aromatic ring -O- and an optionally substituted heterocycle, and are preferably selected from the group consisting of phenyl, 2,6-dimethylphenyl, and 2-oxooxazolidide, and are 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 represented by the formula [P-3]: [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 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 is selected 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 preferably１ , 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 embodiment of the present invention, the onium reagent has the formula [O]: [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 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 is 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, and 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 １－６alkylideneaminooxy, 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, and 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 and DMINTP, more preferably BOI.

[0033] <Base in Step (1)> In one aspect 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 within the range 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 within the range 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 in Step (1)> In one embodiment of the present invention, the solvent used in the 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 of 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 embodiment of the present invention, the condensing agent can be used in an amount within the range 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 aspect 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 within the range 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 in Step (2)> In one embodiment of the present invention, the solvent used in the 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, the reaction time of the step (1) 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]. [In the formula, B Ｐ , Q １ , W, X, G, T, and n are as defined above, and n is an integer from 1 to 50, preferably an integer from 1 to 30, more preferably an integer from 1 to 25, and most preferably an integer from 1 to 15.

[0050] The compound 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: Compounds of formula (I), for example, compounds listed in Table 1 below.

[0053]

[0054] <Method for Obtaining 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]: [In the formula, B Ｐ , Q １ , W, X, L′, and p are as defined above], and the method may comprise: step (1): phosphonation; step (2): hydrolysis; and optionally step (3): ion exchange.

[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]

[0058] <Step (1): Phosphonation> One aspect of the present invention is a compound represented by formula [B-1-0]: [In the formula, B Ｐ , Q １ , W, X, and p are as defined above] with a compound of formula [P-1]: 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 to obtain a compound of formula [B-1'-0]: [In the formula, B Ｐ , Q １ , W, X, R １ and p has the same meaning as defined above.

[0059] <Phosphonylating Reagent in Step (1)> In one embodiment of the present invention, a phosphonylating reagent represented by the formula [P-1]: wherein W is as defined above and R １ and R ２are 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 an 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 aspect 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 Phosphonylating 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 Phosphonylating Reagent Used in Step (1)> In one embodiment of the present invention, the compound of Formula [P-1] can be used in an amount within the range 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 within the range 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 in Step (1)> In one embodiment of the present invention, the solvent used in the 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 of 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 embodiment of the present invention is a compound represented by the formula [B-1′-0]: [In the formula, B Ｐ , Q １ , W, X, R １ and p has the same meaning as defined above] is treated with a hydrolysis solution to obtain a compound of formula [B-1]: [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 ＋ In one embodiment of the present invention, in the 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 ＋ When B is H (aromatic amine), the compound of formula [B-1] is an aliphatic amine salt, a cyclic amine salt, or an aromatic amine salt thereof, for example, a compound of 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. 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 (CH ２ CH ３ ) ３ and -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 in 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 within the range 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 in Step (2)> In one embodiment of the present invention, the solvent used in the 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 in 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 embodiment of the present invention is a compound represented by the formula [B-1′-0]: [In the formula, B Ｐ , Q １ , W, X, R １ and p has the same meaning as defined above] is treated with a hydrolysis solution to obtain a compound of formula [B-1']: [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 of formula [B-1] is further treated with a hydrolysis solution to obtain a compound of formula [B-2]: [In the formula, B Ｐ , Q １ , W, X, and p are as defined above; L' is -O － N ＋ H (cyclic amine), and —O － N ＋ H (cyclic amine) is, for example, —O － (HDBU) ＋ The method includes the step of obtaining a compound of the formula:

[0075] <Hydrolysis Solution in 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 within the range 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 in Step (3)> In one embodiment of the present invention, the solvent used in the 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 One embodiment of the present invention is a method for removing Q from a compound of formula [C-1]. １ to obtain a compound of formula [C-0-1]: [In the formula, B Ｐ , W, X, G, T, n, and p are as defined above; 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]: [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above] is treated with an acid alone or an acid and a scavenger to obtain a compound of formula [C-0-1]: [In the formula, B Ｐ , W, X, G, T, n, and p are as defined above.

[0082] <The above-mentioned acid: Q １Removal Reagent> 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 before use.

[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-mentioned acid: Q １ Amount of Removal Reagent Used> In one embodiment of the present invention, the acid can be used in an amount within the range 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-mentioned acid: Q １ Amount of Removal Reagent Used> In one aspect 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 embodiment of the present invention, １ The solvent used in the step of <Q removal> 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. １Examples of the solvent used in the step <Q removal> 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 embodiment of the present invention, １ The solvent used in the step (3) 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 Reagent> In one aspect 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 Scavenger Used> In one embodiment of the present invention, the scavenger can be used in an amount within the range 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] <The above Q １ Removal Reaction Temperature> In one embodiment 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] <The above Q １ Removal Reaction Time> In one embodiment of the present invention, １The 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], which is 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 present 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], wherein Q １ to obtain a compound of formula [A-1]: [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 represented by the formula [A-0]: [In the formula, B Ｐ , Q １ , W, X, G, T, and n are as defined above] is treated with an acid alone or an acid and a scavenger to obtain a compound of formula [A-1]: [In the formula, B Ｐ, W, X, G, T, and n are as defined above.

[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 Group> One embodiment 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]: [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above.

[0098] One aspect of the present invention is a compound represented by formula [C-1]: [In the formula, B Ｐ , Q １ , W, X, and p are as defined above; G is (1) a silicon substituent; (2) an 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) a group represented by the following general formula [7]: (wherein * represents the bonding position with T; Z is (1) (a soluble polymer that dissolves in an organic solvent)-oxy, (2) (a soluble polymer that dissolves in an organic solvent)-amino, (3) a long-chain alkyloxy, a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or a benzyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, (4) a solid phase support, or (5) a group represented by the following general formulas [8A] to [8N]: (wherein * represents the bonding position with L; j represents an integer of 0 to 4; k represents an integer of 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 represent the following general formulas [9A] to [9E]: (wherein * represents the bonding position; and R ９ represents a long chain alkyl and / or long chain alkenyl; 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 represents (1) a long chain alkyl, (2) a long chain alkyl-carbonyl, or (3) a benzoyl substituted with 1 to 5 long chain alkyloxy and / or long chain alkenyloxy; and R ８ｆ represents a substituent represented by the general formula

[10] : (1) long-chain alkyl, (2) long-chain alkyl-carbonyl, or (3) long-chain alkenyl-carbonyl; and L represents a substituent represented by the general formula

[10] : (Wherein, * represents the bonding position to Z; ** represents the bonding position to 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]: (Wherein, * represents the bonding position to Z; ** represents the bonding position to T; and L １ is optionally substituted C １－１０ Alkylene or optionally substituted C ６－１０ represents an arylene; represents a substituent represented by the following formula: (I) represents a substituent represented by the following formula: (II ... (wherein X and W are as defined above; * represents the bonding position to **O, *O or *N in the above general formulae [4a] to [4d]; ** represents the bonding position to G; and q represents an integer of 0 to 10), but when G is a silicon substituent, T is a single bond. ] is treated under conditions for removing a protecting group for a hydroxyl group to obtain a compound of formula [C-0-2]: [In the formula, B Ｐ , Q １ , W, X, n, and p are as defined above.

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

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

[10] : (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 L represents a substituent represented by the general formula

[10] : (Wherein, * represents the bonding position to Z; ** represents the bonding position to 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]: (Wherein, * represents the bonding position to Z; ** represents the bonding position to T; and L １ is optionally substituted C １－１０ Alkylene or optionally substituted C ６－１０ represents an arylene; represents a substituent represented by the following formula: (I) represents a substituent represented by the following formula: (II ... (wherein X and W are as defined above; * represents the bonding position to **O, *O or *N in the above general formulae [4a] to [4d]; ** represents the bonding position to G; and q represents an integer of 0 to 10), but when G is a silicon substituent, T is a single bond.

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

[0102] <Conditions for Removal of 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 a group in which G is (1) the 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 selected from the group consisting of: (2) optionally substituted C １－１８ Alkyl-carbonyl, optionally substituted C １－１８ (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, 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 present invention, G is (4) the general formula [7]: (wherein Z, L, and * are as defined above) and T is a single bond, the conditions for removing the hydroxyl-protecting group include, for example, treatment with (1) aqueous ammonia, (2) aqueous ammonia / ethanol, or (3) a mixed solution of aqueous ammonia and aqueous methylamine solution.

[0107] <Conditions for Removal of Hydroxyl-Protecting Group: Amount of Reagent Used> In one embodiment of the present invention, the reagent can be used in an amount within the range 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 Removal of Hydroxyl Protecting Group: Reaction Solvent> In one embodiment of the present invention, the solvent used in the <Removal of 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. In one embodiment of the present invention, examples of the solvent used in the <Removal of 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; and halogenated solvents. These solvents may be used in combination.

[0109] <Conditions for Removal of Hydroxyl-Protecting Group: Reaction Temperature> In one embodiment of the present invention, the reaction temperature for the <Removal of 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 Removal of Hydroxyl-Protecting Group: Reaction Time> In one embodiment of the present invention, the reaction time for the <Removal of Q1> 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] <Regarding the final deprotection and isolation step of nucleic acid compound> When compound [C-1] has protecting groups in its molecule, a compound from which all protecting groups have been removed can be produced by carrying out a deprotection treatment according to the type or properties of the protecting groups. For example, all protecting groups possessed by the compound can be removed according to the deprotection method described in "Green's PROTECTIVE GROUPS in ORGANIC SYNTHESIS, 4th Edition, 2006". Specifically, for example, by treating compound [C-1] with (1) aqueous ammonia, (2) aqueous ammonia / ethanol, or (3) a mixed solution of aqueous ammonia and aqueous methylamine, the protecting groups of the substituent [6] possessed in the molecule of compound [C-1] and the amino or hydroxyl group of the nucleic acid base can be removed. In addition, for example, the protecting groups of the substituent [6] possessed in the molecule of compound [C-1] and the amino or hydroxyl group of the nucleic acid base can be removed by treating compound [C-1] with, for example, ammonia water, (2) aqueous ammonia / ethanol, or (3) a mixed solution of aqueous ammonia and aqueous methylamine. 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 prepared by diluting hydrochloric acid or acetic acid in an appropriate solvent.

[0112] <Regarding the purification and separation step> The compound [C-1] from which all of the protecting groups have been removed can be separated and purified from the reaction mixture by a conventional separation and purification method, for example, extraction, concentration, neutralization, filtration, centrifugation, recrystallization, C ８ From C １８ The compound 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 purifying the desired compound using reverse-phase chromatography, a mixed solution of 20 mM triethylamine / acetic acid buffer and acetonitrile can be used as an elution solvent. When purifying the desired compound using ion exchange chromatography, a mixed solution of 1 M saline and 10 mM aqueous sodium hydroxide, or 0.3 M saline in 50 mM phosphate buffer can be used.

[0113] The present invention will be described in more detail below with reference to 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). The "conversion yield (%)" refers to the percentage of the raw material converted into the target product, and is calculated by the following formula: {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 subjected to HPLC analysis under the following conditions. The conversion efficiency was calculated by using the integrated value of the peak area obtained by HPLC absorption at UV = 264 nm. <ODS conditions 1> Column: Waters 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 conditions 2> Column: Waters XBridge C8 (5μm, 4.6 x 75mm), 60°C Detection wavelength: 264nm Mobile phase A: 50mM TEAAaq Mobile phase B: MeOH Flow rate: 0.75mL / min Gradient: 70-95%B (0-20min) 95%B (20-26min) 75%B (26-35min) <LC / MS conditions 1> Equipment used Ultra high performance fluid chromatograph ACQUITY UPLC (waters) Quadrupole time-of-flight mass spectrometer SYNAPT-MS (waters) Column: ACQUITY UPLC BEH C18 1.7μm,2.1 x 50mm (waters) Temperature: 50°C Flow rate: 0.2mL / min Mobile phase A: 10mM aqueous ammonia Mobile phase B: MeCN Gradient: 50-95%B (4min) Detector 1: UV264nm Detector 2: Quadrupole time-of-flight mass spectrometer Ionization method: ESI+ Measurement range: 100-2000 m / z <LC / MS conditions 2> Equipment used Ultra high performance fluid chromatograph ACQUITY UPLC (waters) Quadrupole time-of-flight mass spectrometer SYNAPT-MS (waters) Column: ACQUITY UPLC BEH C18 1.7μm, 2.1×50mm (waters) Temperature: 50℃ Flow rate: 0.2mL / min Mobile phase A: 10mM ammonia water Mobile phase B: MeOH Gradient: 70-95% B (4 min) Detector 1: UV 264 nm Detector 2: Quadrupole time-of-flight mass spectrometer Ionization method: ESI+ Measurement range: 100-2000 m / z <LC / MS condition 3> Instrument used: Agilent 6100 series single quadrupole LC / MS system (Agilent) Column: Waters XBridge C18 (3.5 μm, 4.6 × 150 mm) Temperature: 35°C Flow rate: 1.0 mL / 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 [(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")

[0115] Step 1: Preparation of 4-(octadecylamino)-4-oxobutanoic acid (hereinafter referred to as "G1-suc") Succinic anhydride (8.96 g, 1.1 eq) and triethylamine (17 mL, 1.5 eq.) were added to a solution of octadecane-1-amine (21.94 g) in dichloromethane (500 mL), and the mixture was 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 by suction, washed with acetone (400 mL), and then dried under reduced pressure at 30°C for 3 hours to obtain G1-suc (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") 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (8.56 g, 1.2 eq.) was added to a solution of G1-suc (14.4 g) in tetrahydrofuran (150 mL), and the mixture was stirred at room temperature. Thereafter, 18 g (1.0 eq.) of 1-((2R,6S)-6-(hydroxymethyl)-4-tritylmorpholin-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (hereinafter referred to as "morT-OH") and 4.57 g of 4-(N,N-dimethylamino)pyridine were added, and the mixture was stirred on a 70°C water bath for 1 hour. After cooling to room temperature, 0.1 M aqueous sodium dihydrogen phosphate solution was added and stirred for a while, after which the aqueous layer was removed, and the organic layer was washed once with 0.1 M aqueous sodium dihydrogen phosphate solution and once with saturated saline diluted two-fold with water. The aqueous layers were combined and extracted with dichloromethane, and the organic layers were combined and dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated and the residue was dried under reduced pressure to give [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methyl 4-(octadecylamino)-4-oxobutanoate (hereinafter referred to as "G1-suc-morT-ON") (white amorphous, 28.1 g, 89.5%). This was dissolved in 140 mL of dichloromethane, and while stirring on 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 completion of the dropwise addition, the reaction solution was poured into a solution cooled by adding ice to 100 mL of saturated aqueous sodium bicarbonate, while stirring. After confirming that the pH of the aqueous layer was 7 to 8, the aqueous layer was extracted with dichloromethane. The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by column chromatography on silica gel using a dichloromethane-methanol mixture as a mobile phase, and dried under reduced pressure to obtain 19.89 g of G1-suc-morT-OFF as a powder.

[0117] １ H-NMR (CDCl ３ ): δ 8.90 (1H, bs); 7.25 (1H, d, J = 1.6 Hz); 5.72 (1H, dd, J = 9.6 Hz, 2.6 Hz); 5.65 (1H, m); 4.14 (2H, d, J = 5.2 Hz); 3.98 (1H, m); 3.23 (2H, dd, J = 12.8 Hz, 7.0 Hz); 3.12 (2H, dd, J = 12 Hz, 2.6 Hz); 2.95 (2H, dd, J = 12.8 Hz, 1.8 Hz); 2.60 - 2.75 (4H, m); 2.47 (2H, t, J = 6.8 Hz); 1.95 (3H, d, J = 1.6 Hz); 1.48 (2H, m), 1.21 - 1.34 (29H, m); 0.88 (3H, t, J = 6.4 Hz) ESI-MS(+): 593.36 (M + H) (LC / MS condition 1)

[0118] Reference Example 1-1 In the same manner as in Reference Example 1, (1) <4-(octadecylamino)-4-oxobutanoate [(2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl]methyl> (G1-suc-morA-ON), <4-(octadecylamino)-4-oxobutanoate [(2S,6R)-6-(6-benzamido-9H-purin-9-yl)morpholin-2-yl]methyl> (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-oxobutanoate [(2S,6R)-6-(6-(2-cyanoethoxy)-2-(phenoxyacetamido)-9H-purin-9-yl)-4-tritylmorpholin-2-yl]methyl> (G1-suc-morG-ON), <4-(octadecylamino)-4-oxobutanoate [(2S,6R)-6-(6-(2-cyanoethoxy)-2-(phenoxyacetamido)-9H-purin-9-yl)morpholin-2-yl]methyl> (G1-suc-morG-OFF) were prepared.

[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: Production 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 the addition of 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 completion of the reaction, a 1 M aqueous solution of sodium dihydrogen phosphate was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated to give G2-suc (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 the addition of 500 mg (1.03 mmol) of morT-OH and 132 mg (1.09 mmol) of 4-(N,N-dimethylamino)pyridine, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, 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 removed by evaporation. The resulting residue was purified by silica gel chromatography to give G2-suc-morT-ON (1.09 g, 89%).

[0122] １ H-NMR (CDCl ３): δ8.04 (1H, s); 7.17 to 7.51 (15H, m); 6.98 (1H, s); 6.12 (1H, dd, J = 9.6Hz, 2.4Hz) ; 5.25 (1H, m); 4.34-4.37 (1H, m); 4.26-4.30 (2H, m); 4.11-4.16 (2H, m); 4.00-4.0 8 (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 to 1.44 (2H, m); 1.24 (60H, m); 0.87 (6H, t, J = 6.8Hz)

[0123] Step 3: Preparation 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 the mixture was 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 G2-suc-morT-OFF (373 mg, 95%).

[0124] １ H-NMR (CDCl ３ ): δ8.04 (1H, bs); 7.24 (1H, s); 5.70 (1H, d, J=2Hz); 5.21 to 5.26 (1H, m ); 4.28-4.31 (2H, m); 4.13-4.17 (4H, m); 3.96-4.00 (1H, m); 3.11 (1H, dd, J=12.4, 2Hz); 2.94 (1H, dd, J=12.8, 2.4Hz); 2.57 to 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") Using 2-hydroxypropane-1,3-diyl dioleate as a starting material, 4-((1,3-bis(oleoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (hereinafter referred to as "G3-suc") was prepared in the same manner as in Step 1 of Reference Example 2. Then, G3-suc-morT-ON was prepared in the same manner as in Step 2 of Reference Example 2.

[0127] １ H-NMR (CDCl ３ ): δ8.00 (1H, s); 7.17 to 7.51 (15H, m); 6.99 (1H, s); 6.09 to 6.12 (1H, m); 5.29 to 5.38 (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 = 7.2Hz) ); 1.97 to 2.02 (8H, m); 1.83 (3H, s); 1.57 to 1.61 (2H, m); 1.28 (44H, m); 0.89 (6H, t, J = 6.8Hz)

[0128] Step 2: Preparation of G3-suc-morT-OFF G3-suc-morT-OFF was prepared in the same manner as in Step 3 of Reference Example 2.

[0129] １ H-NMR (CDCl ３):δ7.97(1H,bs);7.24(1H,s);5.69~5.72(1H,m);5.29~5.38(4H,m);5.21~5.25(1H,m);4.27~4.31(2H,m);4.13~4.17(4H,m);3.97~3.99(1H,m);3.11(1H,d,J=12Hz);2.94(1H,d,J=13.2Hz);2.57~2.67(4H,m)2.31(4H,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: Preparation 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, followed by 1.45 mL (10.7 mmol) of triethylamine and 1 g (5.37 mmol) of tert-butyl piperazine-1-carboxylate, and the mixture was stirred at room temperature for 16 hours. 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 4-stearoylpiperazine-1-carboxylate tert-butyl (1.64 g; 67%). 18 mL of dichloromethane was added to the mixture, and the mixture was stirred at 0°C. 2.77 mL (36.2 mmol) of trifluoroacetic acid was added at 0°C, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, a saturated aqueous solution of 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 completion of the reaction, 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 555 mg (2.90 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, and the mixture was stirred at 70° C. Subsequently, morT-OH (1 g, 2.07 mmol) and 265 mg (2.17 mmol) of 4-(N,N-dimethylamino)pyridine were added, and the mixture was stirred at 70° C. for 30 minutes. After the reaction was completed, the reaction mixture was cooled to room temperature, and 0.1 M aqueous sodium dihydrogen phosphate solution was added to the reaction mixture. 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 give G4-suc-morT-ON (1.68 g, 89%).

[0132] １ H-NMR (CDCl ３ ): δ8.00 (1H, s); 7.16 to 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); 3.44-3.4 8 (4H, m); 3.32 to 3.34 (1H, m); 3.09 to 3.12 (1H, m); 2.60 to 2.64 (4H, m); 2.3 1 (2H, t, J = 7.6Hz); 1.82 (3H, s); 1.23 to 1.42 (32H, m); 0.86 (3H, t, J = 6.8)

[0133] Step 3: Preparation of G4-suc-morT-OFF Preparation was carried out in the same manner as in Step 3 of Reference Example 2.

[0134] １ H-NMR (CDCl ３): δ8.32 (1H, bs); 7.23 (1H, s); 5.67 to 5.70 (1H, m); 4.12 to 4.19 (2H, m); 3.96 to 4.01 (1H, m); 3.47 to 3.67 (8H, m) 3.10 to 3.13 (1H , m); 2.95 to 2.98 (1H, m); 2.60 to 2.72 (4H, m); 2.33 (2H, t, J = 7.2Hz); 1.95 (3H, s); 1.25 to 1.31 (32H, m); 0.88 (3H, t, J = 7.6Hz)

[0135] Reference Example 5: 4-(octadecylcarbamoyl)benzoate [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl (hereinafter referred to as "G5-tpa-morT-OFF") Step 1: Preparation of 4-(octadecylcarbamoyl)benzoate [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methyl (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 (CDCl ３ ): δ8.24 (1H, s); 7.97 (2H, d, J = 8Hz); 7.77 (2H, d, J = 8Hz); 7.17 to 7.46 (15H, m); 6.95 (1H, s); 6.12 to 6.16 (1H, m); 4.49 to 4.51 (1H, 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: Preparation of G5-tpa-morT-OFF Preparation was carried out in the same manner as in Step 3 of Reference Example 2.

[0138] １ H-NMR (CDCl ３): δ8.24 (1H, bs); 8.11 (2H, d, J = 8.4Hz); 7.84 (2H, d, J = 8.4Hz); 7.24 (1H, s); 6.14 to 6.17 (1H, m); 5.74 to 5.77 (1H, m); 4.40 to 4.45 (2H, m) ; 4.13 to 4.19 (1H, m); 3.45 to 3.50 (2H, m); 3.14 to 3.18 (1H, m); 3.05 to 3.08 (1H, m); 1.93 (3H, s); 1.26 to 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 prepared 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") Preparation was carried out in the same manner as in Step 2 of Reference Example 2.

[0141] １ H-NMR (CDCl ３ ): δ8.08 (1H, bs); 7.81 (2H, d, J = 7.6Hz); 7.16 to 7.50 (17H, m); 6.97 (1H, s); 6.08 to 6.10 (1H, m); 4.33 to 4.39 (1H, m); 4.02 to 4.0 4 (2H, m); 3.31 to 3.79 (11H, m); 3.08 to 3.11 (1H, m); 2.60 to 2.69 (4H, m); 1.81 (3H, s); 1.23 to 1.44 (34H, m); 0.86 (3H, t, J = 6.4Hz)

[0142] Step 3: Preparation 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 the mixture was 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 (CDCl ３ ): δ8.05 (1H, bs); 7.79 (2H, d, J = 7.6Hz); 7.45 (2H, d, J = 7.6Hz); 7.23 (1 H, 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 to 3.79 (8H, m); 3.08 to 3.11 (1H, m); 2.93 to 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 prepared in the same manner as in Step 2 of Reference Example 2, using 3,4,5-trioctadecoxybenzoic acid.

[0146] １ H-NMR (CDCl ３): δ7.90 (1H, bs); 7.12 to 7.45 (17H, m); 6.97 (1H, s); 6.12 to 6.14 (1H, m); 4.46 to 4.51 (1H, m); 4.28 to 4.32 (1H, m); 4.16 to 4.20 (1H , m); 3.90 to 4.00 (6H, m); 3.37 to 3.40 (1H, m); 3.22 to 3.25 (1H, m); 1.78 to 1.82 (5H, m); 1.23 to 1.50 (96H, m); 0.86 (9H, t, J = 6.8Hz)

[0147] Step 2: Preparation of G7-morT-OFF G7-morT-OFF was prepared in the same manner as in Step 3 of Reference Example 2.

[0148] １ H-NMR (CDCl ３ ): δ7.98 (1H, bs); 7.22 (3H, m); 5.69 to 5.72 (1H, m); 4.32 to 4.36 (2H, m); 4.08 to 4.12 (1H, m); 3.94 to 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 8.1 mL of chloroform was added to 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, followed by 120 mg (1.90 mmol) of ethylene glycol, and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, 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 2-hydroxyethyl 3,4,5-trioctadecyloxybenzoate (882 mg; 56%).

[0151] １ H-NMR (CDCl ３ ): δ7.26 (2H, s); 4.45 to 4.47 (2H, m); 3.95 to 4.03 (8H, m); 1.25 to 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 the same manner as in Step 1 of Reference Example 2, and then G8-suc-morT-ON was obtained in the same manner as in Step 2 of Reference Example 2.

[0153] １ H-NMR (CDCl ３): δ7.87 (1H, bs); 7.12 to 7.43 (17H, m); 6.97 (1H, s); 6.07 to 6.10 (1H, m); 4.33 to 4.46 (5H, m); 3.91 to 4.07 (8H, m); 3.31 ~3.34 (1H, m); 3.07 to 3.10 (1H, m); 2.56 to 2.60 (4H, m); 1.68 to 1.80 (5H, m); 1.23 to 1.50 (96H, m); 0.86 (9H, t, J=7.2Hz)

[0154] Step 3: Production 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 (CDCl ３ ): δ7.96 (1H, bs); 7,23 (3H, m); 5.67 to 5.69 (1H, m); 4.40 to 4.47 (5H, m); 3.94 to 4.11 (8H, m); 3.09 to 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 4-(Dioctadecylamino)-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl ester (hereinafter referred to as "G9-suc-morT-OFF") Step 1 Preparation of 4-(dioctadecylamino)-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl ester (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 (CDCl ３): 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); 1.8 2 (3H, s); 1.23~1.52 (64H, m); 0.85 (6H, t, J=6.8Hz)

[0158] Process 2 G9-suc-morT-OFF manufacturing reference example 2 and process 3 are manufactured using the same method.

[0159] １ H-NMR (CDCl ３ ): 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: 4-[{1-(octadecylamino)-1-oxo-3-phenylpropan-2-yl}amino]-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl ester (hereinafter referred to as "G10-suc-morT-OFF"). Step 1: Preparation of 4-[{1-(octadecylamino)-1-oxo-3-phenylpropan-2-yl}amino]-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl ester (hereinafter referred to as "G10-suc-morT-ON"). To 500 mg (1.88 mmol) of 2-tert-butoxycarbonylamino-3-phenyl-propanoic acid was added 9.4 mL of tetrahydrofuran, 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, and 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, 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-butoxycarbonylamino-N-octadecyl-3-phenyl-propanamide (779 mg, 80%).

[0161] １ H-NMR (CDCl ３ ): δ7.18 to 7.28 (5H, m); 5.57 (1H, bs); 5.06 (1H, bs); 4.20 to 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 was added 15 mL of dichloromethane, 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. Then, a reaction similar to that in Step 2 of Reference Example 2 was performed to produce G10-suc-morT-ON.

[0163] １ H-NMR (CDCl ３ ): δ8.08 (1H, bs); 7.16 to 7.32 (20H, m); 6.98 (1H, s); 6.29 to 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 (1 H, m); 4.02 (2H, d, J = 5.6Hz); 3.73 to 3.77 (1H, m); 2.92 to 3.33 (6H, m); 2.3 9 to 2.67 (4H, m); 1.84 (3H, s); 1.21 to 1.45 (32H, m); 0.88 (3H, t, J = 7.6Hz)

[0164] Step 2: Preparation 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 (CDCl ３ ): δ8.33 (1H, bs); 7.16 to 7.32 (6H, m); 6.38 to 6.40 (1H, m); 5.67 to 5.71 (2H, m); 4.54 to 4.58 (1H, m); 4.08 to 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 formulae of the compounds described in Reference Examples 1 to 10 above are shown in Table 3 below.

[0167]

[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) 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. To this solution, 481 μL (2.51 mmol) of diphenyl phosphite was added and the mixture was stirred at room temperature for 2 hours. After 2 hours, 100 mL of 1 M triethylammonium bicarbonate solution was added and the mixture was 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 condition 1). １ H-NMR (CDCl ３): 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, 7.3 Hz, 6H); 7.19 (d, J = 7.3, 7.3 Hz, 3H); 6.40 (dd, J = 9.6, 2.3 Hz, 1H); 4.43 to 4.49 (m, 1H) ), 3.81-3.95 (m, 2H); 3.50 (ddd, J = 11.0, 2.3, 2.3 Hz, 1H); 3.27 (ddd, J = 11.9, 2.3, 2.3 Hz, 1H); 2.95 (q, J = 7.3 Hz, 7.2H); 1.81 (dd, J = 10.0, 10.0 Hz, 1H); 1.61 (dd, J = 11.0, 11.0 Hz, 1H); 1.23 (t, J = 7.3 Hz, 10.8H). Ｂｚ 500 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 (CDCl ３ ): 9.01 (brs, 1H); 8.80 (s, 1H); 8.61 to 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, 1.8H z, 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, 1H); 4.4 4 to 4.49 (m, 1H); 3.82 to 3.97 (m, 2H); 3.49 (ddd, J = 11.0, 2.3, 2.3Hz, 1H); 3.34 to 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 to 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).

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

[0172] Example 2-1: H-phosphonate-PMO[C Ｂｚ -A Ｂｚ ]-ON 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. To this solution, 148 μL (0.739 mmol) of 5 M sodium methoxide / methanol solution was added 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 saline and saturated aqueous sodium bicarbonate. The organic layer was dried over sodium sulfate, and the solvent was distilled off. The resulting residue was dissolved in dichloromethane and reprecipitated with diisopropyl ether to obtain HO-PMO[C Ｂｚ -A Ｂｚ ]-ON was obtained (3.67 g, 96%). Ｂｚ -A Ｂｚ1.5 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 condition 1).

[0173] Example 2-2: H-phosphonate-PMO[C Ｂｚ -A Ｂｚ -T-C Ｂｚ Preparation of [-T]-ON AcO-PMO[C Ｂｚ -A Ｂｚ -T-C Ｂｚ -T]-ON is AcO-PMO[C Ｂｚ ]-ON by a known method. Ｂｚ -A Ｂｚ -T-C Ｂｚ 4 g (1.88 mmol) of [-T]-ON was suspended in 38 mL of tetrahydrofuran. To this solution, 450 μL (2.26 mmol) of 5 M sodium methoxide / methanol solution was added 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 distilled off. The resulting residue was dissolved in dichloromethane and reprecipitated with diisopropyl ether to obtain HO-PMO[C Ｂｚ -A Ｂｚ -T-C Ｂｚ -T]-ON was obtained (3.92 g, 99%). Ｂｚ -A Ｂｚ -T-C Ｂｚ6.2 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 -T-C Ｂｚ -T]-ON (triethylamine salt) was obtained (6.56 g, 98%). 1 g of the obtained 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 -T-C Ｂｚ 960 mg of [-T]-ON (DBU salt) was obtained. ESI-MS (+): 2158.7 (LC / MS condition 2).

[0174] Example 2-3: H-phosphonate-PMO[C Ｂｚ -G ＣＥ，Ｐａｃ -A Ｂｚ -A Ｂｚ -G ＣＥ，Ｐａｃ ]-ON Preparation of 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，Ｐａｃ19 g (7.07 mmol) of HO-PMO[C]-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 distilled off. The resulting residue was purified by silica gel chromatography to obtain HO-PMO[C]-ON. 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 obtained 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 490 mg of 4-ON (DBU salt) was obtained. ESI-MS (+): 2606.9 (LC / MS condition 2).

[0175] Example 2-4: H-phosphonate-PMO[G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -TTC Ｂｚ]-ON TrocO-PMO[G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -TTC Ｂｚ 1 g (0.40 mmol) of HO-PMO[G]-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 (4.01 mmol) of zinc powder were added and stirred for 1 hour. After 1 hour, the reaction solution was filtered through Celite, and the filtrate was evaporated to remove the solvent. 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%). 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 condition 3).

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

[0177] Example 3: PMO Oligomer Preparation

[0178] Example 3-1: G1-suc-PMO[C Ｂｚ -A Ｂｚ ]-ON G1-suc-PMO[C Ｂｚ]-OFF 17 mg (0.025 mmol) and H-phosphonate-PMO[A 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, followed by stirring at room temperature. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After completion of the reaction, 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: 11.4 min, target product Rt: 17.2 min, conversion yield 92.1%, ODS condition 1). ESI-MS (+): 1367.8 (M+H) (LC / MS condition 1).

[0179] Example 3-2: G1-suc-PMO[C Ｂｚ -A Ｂｚ ]-ON 2 mL of solution a shown below, delivered at a flow rate of 0.25 mL / min, and 2 mL of solution b shown below, delivered 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 completion of the reaction, 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 solution 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 Ｂｚ 119 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 -CＢｚ ]-ON 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, followed by stirring at room temperature. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After completion of the reaction, 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: 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 condition 1).

[0181] Example 3-4: TBSO-PMO[C Ｂｚ -C Ｂｚ ]-ON TBSO-PMO[C Ｂｚ ]-OFF 11 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, followed by stirring at room temperature. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After completion of the reaction, 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: 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 condition 1).

[0182] Example 3-5: G1-suc-PMO[T-G ＣＥ，Ｐａｃ -AＢｚ ]-ON G1-suc-PMO[T-G 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 the mixture was stirred at 40°C. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After completion of the reaction, 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: 12.9 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] Example 3-6: G1-suc-PMO[T-G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -A Ｂｚ ]-ON G1-suc-PMO[T-G 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 the mixture was stirred at 40°C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidation 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 mixture, 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: 13.6 min, target product Rt: 19.5 min, conversion yield 91.6%, ODS condition 2). ESI-MS (+): 2362.0 (LC / MS condition 2).

[0184] Example 3-7: G1-suc-PMO[T-G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -A Ｂｚ -A Ｂｚ ]-ON Production of G1-suc-PMO[T-G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -A Ｂｚ ]-OFF 67 mg (0.0251 mmol) and H-phosphonate-PMO[A Ｂｚ 29 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 the mixture was stirred at 40°C. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After completion of the reaction, 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: 14.4 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 -T-C Ｂｚ -C Ｂｚ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -TTC Ｂｚ -A Ｂｚ ]-ON (using diphenyl chlorophosphate as a condensation agent) Ｂｚ -C Ｂｚ -T-C 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 the mixture was stirred at 40°C. After 10 minutes, 100 μL of the reaction mixture was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After the reaction was completed, 10% aqueous sodium thiosulfate solution was added to the reaction mixture, 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: 16.1 min, target product Rt: 19.5 min, conversion yield 94.4%, ODS condition 2). ESI-MS (+): 5117.9 (LC / MS condition 2).

[0186] Example 3-9: G1-suc-PMO[C Ｂｚ -C Ｂｚ -T-C Ｂｚ -C Ｂｚ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -TTC Ｂｚ -A Ｂｚ ]-ON (using bromotripyrrolidinophosphonium hexafluorophosphate as a condensation agent) Ｂｚ -C Ｂｚ -T-C 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, followed by stirring at 40°C. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After completion of the reaction, 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: 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 -T-C Ｂｚ -C Ｂｚ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -TTC Ｂｚ -A Ｂｚ ]-ON (using diphenylbis(2,6-dimethylphenyl) chlorophosphate as a condensation agent) Ｂｚ -C Ｂｚ -T-C 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 the mixture was stirred at 40°C. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After completion of the reaction, 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: 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 -T-C Ｂｚ -C Ｂｚ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -TTC Ｂｚ -A Ｂｚ ]-ON (using bis(2-oxo-3-oxazolidinyl)phosphinic chloride as a condensation agent) Ｂｚ -C Ｂｚ -T-C 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, followed by stirring at 40°C. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After completion of the reaction, 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: 16.1 min, target product Rt: 19.5 min, conversion yield 95.9%, ODS condition 2).

[0189] Example 3-12: G1-suc-PMO[T-A Ｂｚ ]-ON (using piperidine instead of dimethylamine as the oxidizing solution) 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 the mixture was 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 completed, 10% aqueous sodium thiosulfate solution was added to the reaction mixture, 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: 12.0 min, target product Rt: 19.6, 20.1 min, conversion yield 98.9%, ODS condition 2). ESI-MS (+): 1317.7 (LC / MS condition 2).

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

[0191] Example 4-1: G1-suc-PMO[A Ｂｚ -CＢｚ Preparation of -TT]-ON G1-suc-PMO[A Ｂｚ -C Ｂｚ ]-OFF 28 mg (0.025 mmol) and H-phosphonate-PMO[T-T]-ON (triethylamine salt) 37 mg (0.0375 mmol) 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 solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After completion of the reaction, 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: 13.8 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 2 mL of the following solution a, delivered at a flow rate of 0.2 mL / min, and 2 mL of the following solution b, delivered 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, a 10% aqueous solution of sodium thiosulfate was added to the reaction solution, and the organic and aqueous layers were separated. The organic layer was diluted 20 times with an 80% aqueous acetonitrile solution 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: 257 mg (0.263 mmol) of H-phosphonate-PMO[TT]-ON and G1-suc-PMO[A Ｂｚ -C Ｂｚ197 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 G1-suc-PMO[TTA Ｂｚ ]-OFF 34 mg (0.025 mmol) and H-phosphonate-PMO[T-T]-ON (triethylamine salt) 37 mg (0.0375 mmol) 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 solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 10 minutes. After completion of the reaction, 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: 12.4 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[T-T-A Ｂｚ Preparation of -TT]-ON 2 mL of the following solution a, pumped at a flow rate of 0.2 mL / min, and 2 mL of the following solution b, 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, a 10% aqueous solution of sodium thiosulfate was added to the reaction solution, and the organic and aqueous layers were separated. The organic layer was diluted 20 times with an 80% aqueous acetonitrile solution 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[T-T]-ON 257 mg (0.263 mmol) and G1-suc-PMO[T-T-A Ｂｚ 239 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[T-G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -A Ｂｚ -C Ｂｚ -A Ｂｚ -T-C Ｂｚ -T]-ON (using triethylamine salt of H-phosphonate) ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -A Ｂｚ ]-OFF 33 mg (0.0123 mmol) and H-phosphonate-PMO[C Ｂｚ -A Ｂｚ -T-C Ｂｚ40 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 the mixture was stirred at 40°C. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After completion of the reaction, 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: 14.4 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] Example 4-6: G1-suc-PMO[T-G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -A Ｂｚ -C Ｂｚ -A Ｂｚ -T-C Ｂｚ -T]-ON (using DBU salt of H-phosphonate) ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -A Ｂｚ ]-OFF 33 mg (0.0123 mmol) and H-phosphonate-PMO[C Ｂｚ -A Ｂｚ -T-C Ｂｚ40 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, followed by stirring at 40°C. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After completion of the reaction, 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: 14.4 min, target product Rt: 18.6 min, conversion yield 95.8%, target product purity 53.1%, ODS condition 2).

[0197] Example 4-7: G1-suc-PMO[C Ｂｚ -C Ｂｚ -T-C Ｂｚ -C Ｂｚ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -TTC Ｂｚ -C Ｂｚ -G ＣＥ，Ｐａｃ -A Ｂｚ -A Ｂｚ -G ＣＥ，Ｐａｃ ]-ON Preparation of G1-suc-PMO[C Ｂｚ -C Ｂｚ -T-C 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 the mixture was 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 completed, 10% aqueous sodium thiosulfate solution was added to the reaction mixture, 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: 16.1 min, target product Rt: 19.8 min, conversion yield 91.5%, ODS condition 2). ESI-MS (+): 7064.4 (LC / MS condition 2).

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

[0199] Example 5: G1-suc-PMO[C Ｂｚ -C Ｂｚ -T-C Ｂｚ -C Ｂｚ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -TTC Ｂｚ -T-G ＣＥ，Ｐａｃ -A Ｂｚ -A Ｂｚ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -T-G ＣＥ，Ｐａｃ Preparation of -TT]-ON G1-suc-PMO[C Ｂｚ -C Ｂｚ -T-C Ｂｚ -C Ｂｚ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -TTC Ｂｚ ]-OFF 28 mg (0.00631 mmol) and H-phosphonate-PMO[T-G ＣＥ，Ｐａｃ -A Ｂｚ -A Ｂｚ -G ＣＥ，Ｐａｃ -G ＣＥ，Ｐａｃ -T-G ＣＥ，Ｐａｃ43 mg (0.00947 mmol) of [-T-T]-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, followed by stirring at 40°C. After 10 minutes, 100 μL of the reaction solution was mixed with 100 μL of an oxidation solution (0.1 M iodine, 2 M dimethylamine / THF solution) and stirred at room temperature for 5 minutes. After completion of the reaction, 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: 15.2 min, target product Rt: 17.6 min, conversion yield 91.0%, ODS condition 2). 190 μL of a 3:1 mixture of aqueous ammonia and ethanol was added to 10 μL of the reaction solution, and the mixture was stirred at 55° C. for 18 hours. The reaction solution was filtered, and the filtrate was diluted 50 times with 20% aqueous acetonitrile and subjected to MS measurement, confirming the production of the target product. ESI-MS (+): 6848.4 (LC / MS condition 1)

Claims

1. Formula [A-1]: 【Chemistry 1】 [In the formula, B Ｐ are the same or different and are optionally protected nucleobases; 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 is the same or different and is a hydroxyl group substituted with a group removable 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 removable under basic conditions １－６ alkyl)amino, di(amino-C substituted with a group removable under basic conditions １－６ alkyl)amino, or the general formula [2]: 【Chemistry 2】 (In the formula, * indicates the position of bond to P; a is an integer from 0 to 2; E is CH ２ , C-H-A １ or N-A ２ and A １ is C １－６ Alkyl, mono-substituted with groups removable under basic conditions (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 an amino substituted with an amidino substituted with a group removable under basic conditions, or a compound represented by the general formula [3]: 【Chemistry 3】 (In the formula, * indicates the position of attachment to 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-substituted with groups removable under basic conditions (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) a silicon-substituted group, (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, * is the binding position to T; Z is, (1) (Soluble polymer that dissolves in an organic solvent)-oxy, (2) (Soluble polymer that dissolves in an 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]: 【Chemistry 5】 (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 each is a long chain alkyl; R ８ｃ are the same or different and are represented by the general formulas [9A] to [9E]: 【Chemistry 6】 (In the formula, * is the bond position; and R ９ is a long chain alkyl and / or long chain alkenyl) A substituent selected from the group consisting of R ８ｄ are the same or different and each is a hydrogen atom, a halogen, a long-chain alkyl optionally substituted with 1 to 13 halogens, or a long-chain alkyloxy optionally substituted with 1 to 13 halogens; 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 A substituent selected from the group consisting of and L is represented by the general formula [10]: 【Chemistry 7】 (In the formula, * is the bonding position with Z; ** is the position of attachment to T; and L １ is an optionally substituted C ２－１０ Alkylene or optionally substituted C ６－１０ arylene), or general formula [10-1]: 【Chemistry 8】 (In the formula, * is the bonding position with Z; ** is the position of attachment to T; and L １ is an optionally substituted C １－１０ Alkylene or optionally substituted C ６－１０ It is an arylene. is a substituent represented by and 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 position of bond to O; ** is the position of attachment 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]: 【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 is removable 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:

2. Furthermore, the formula [C-1]: 【Chemistry 12】 [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, Formula [C-0-1]: 【Chemistry 13】 [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]: 【Chemistry 14】 [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) Treating with an oxidizing agent and an organic amine, Formula [C-1-1]: 【Chemistry 15】 [In the formula, B Ｐ , Q １ , W, X, G, T, n, and p are as defined above.

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

3. Furthermore, the formula [C-1]: 【Chemistry 16】 [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 17】 [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 18】 [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 is selected from the group consisting of alkyl, optionally substituted phenyl, and PH(=O)-OH. and a base, followed by (2) Treating with a hydrolysis solution; Formula [C-0-2-1]: 【Chemistry 19】 [In the formula, B Ｐ , Q １ , W, X, L′, n, and p are as defined above.

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

4. Furthermore, the formula [C-0-2-1]: 【Chemistry 20】 [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 21】 [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) Treating with an oxidizing agent and an organic amine, Formula [C-1-2]: 【Chemical 22】 [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 23】 [In the formula, B Ｐ , Q １ , W, X, and p are as defined above. The compound (1) Formula [P-1]: 【Chemistry 24】 [Wherein, W is an oxygen atom or a sulfur atom, R １ , and R ２ are each the same or different and selected from the group consisting of H, optionally substituted C 1-6 alkyl, optionally substituted phenyl, and PH(═O)—OH. and a base, followed by (2) Treating with a hydrolysis solution; Formula [B-1]: 【Chemistry 25】 [In the formula, B Ｐ , Q １ , W, X, L′, and p are as defined above.

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

6. Furthermore, the formula [A-0]: 【Chemistry 26】 [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, Formula [A-1]: 【Chemical 27】 [In the formula, B Ｐ , W, X, G, T, and n are as defined above.

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

7. G is of the formula: 【Chemistry 28】 [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] [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, 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 (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 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) no)phosphoryl)morpholin-2-yl)methoxy-(dimethylamino)phosphoryl)morpholin-2-yl)methoxy-(dimethylamino)phosphoryl)-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 The compound of formula [B-1] according to claim 8, selected from the group consisting of:

10. Formula [B-0-1]: 【Chemical 29】 [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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