Method for producing oligonucleotide compound

Abstract

The present invention relates to a method for producing a compound [C] represented by general formula [C] by subjecting a compound [A] having a hydroxyl group or a primary or secondary amino group and a compound [B] represented by general formula [1], which has a phosphorus atom-containing substituent group, to a condensation reaction, the method being characterized by being carried out in the presence of at least one reaction accelerator selected from the group consisting of a quaternary ammonium salt, a quaternary imidazolium salt, a quaternary morpholinium salt, a quaternary phosphonium salt, a quaternary piperidinium salt, a quaternary pyridinium salt, a quaternary pyrrolidinium salt and a quaternary sulfonium salt.

Description

[Technical field]

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

[0002] Known methods for producing oligonucleic acid compounds include the solid-phase method and the liquid-phase method. The solid-phase method is a heterogeneous reaction method in which a nucleic acid is extended while contacting a substrate supported on a solid-phase carrier with a solution containing a reaction reagent. In the solid-phase method, a so-called batch method is used 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). Also known is a pseudo-flow synthesis method in which a solid-phase carrier is placed in a column, as in an automatic nucleic acid synthesizer (e.g., DNA, RNA synthesizer), and a solution containing a reaction reagent is passed through the column to cause a reaction. On the other hand, the liquid phase method is a homogeneous reaction method in which nucleic acids are extended by reacting in a solution containing both a substrate and a reaction reagent. The liquid phase method also uses a batch method in which the reaction is carried out in a container (see, for example, Patent Document 2 and Patent Document 3).

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

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

[0005] On the other hand, as homogeneous reaction methods, synthesis methods using liquid phase methods and hydrophobic group-bound nucleosides or pseudo solid-phase protected nucleosides are known. The liquid phase method is a homogeneous reaction method in which the reaction is carried out in a solution containing both the substrate and the reaction reagent. Compared to the solid phase method, the reaction efficiency is higher and the reaction rate is faster, but column purification is required to remove impurities such as the reaction reagent and reaction solvent. The synthesis method using hydrophobic group-bonded nucleosides or pseudo solid-phase protected nucleosides can react in a homogeneous system, similar to the liquid phase method, and therefore has higher reaction efficiency and faster reaction rate than the solid phase method. 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). In these homogeneous reaction methods, a nonpolar solvent such as chloroform is used in the condensation reaction. However, as reported in the synthesis of morpholino nucleic acid (see, for example, Patent Document 5), the condensation reaction in a nonpolar solvent requires a very long time, and therefore the use of a nonpolar solvent in a homogeneous reaction is not preferable in terms of improving the reaction efficiency and reaction rate of the condensation reaction. [Prior art documents] [Patent documents]

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

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

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

[0009] The present inventors have found that phosphorus bonds can be formed efficiently by using a reaction promoter in the condensation reaction of an oligonucleic acid compound, and have completed the present invention.

[0010] The present invention relates to a compound [A] having a hydroxyl group or a primary or secondary amino group and a compound represented by the following general formula [1]: [ka] [In the formula, ** indicates the bond position with the residue of compound [B]; D is a halogen, a 5- to 6-membered saturated cyclic amino or a di(C 1-6 (alkyl)amino; W 0represents a lone pair, an oxygen atom, or a sulfur atom; and X is a hydroxyl group substituted with a group that can be removed under neutral conditions, 1,1,3,3-tetra(C 1-6 Alkyl) guanidyl, C 1-6 Alkoxy, di(C 1-6 Alkyl)amino, mono(amino-C substituted with a group removable 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 the following general formula [2]: [ka] (In the formula, * indicates the bond position with P; a represents an integer from 0 to 2; E is CH 2 , C.H.-A. 1 or NA 2 represents; A 1 is C 1-6 Alkyl, monosubstituted 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 Ammonio, amino, or the following general formula [3]: [ka] (In the formula, * indicates the bond position to E; b represents an integer of 0 to 2; c represents 0 or 1; R 11is C 1-6 represents alkyl; M is CH 2 , an oxygen atom, a sulfur atom, or N-(a group that can be removed under basic conditions). (hereinafter referred to as "substituent [3]"); and A 2 is C 1-6 Alkyl, monosubstituted 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 represents an alkyl group, a group that can be removed under basic conditions, an aryl group, or a heteroaryl group. (hereinafter referred to as "substituent [2]"). A compound [B] having a substituent containing a phosphorus atom represented by the following general formula [C] (hereinafter referred to as "substituent [1]") is subjected to a condensation reaction to obtain a compound represented by the following general formula [C]: [ka] (In the formula, W 0 and X is as defined above; A represents a residue obtained by removing one hydrogen atom from a hydroxyl group or a primary or secondary amino group of the compound [A]; and B represents the residue obtained by removing the substituent [1] from compound [B]. (hereinafter referred to as "compound [C]"), which is characterized in that the method is carried out in the presence of at least one reaction promoter selected from the group consisting of quaternary ammonium salts, quaternary imidazolium salts, quaternary morpholinium salts, quaternary phosphonium salts, quaternary piperidinium salts, quaternary pyridinium salts, quaternary pyrrolidinium salts, and quaternary sulfonium salts. Effect of the Invention

[0011] An oligonucleic acid compound is a compound having a structure in which two or more nucleoside units are linked together via phosphorus bonds. In order to produce an oligonucleic acid compound, it is necessary to carry out a condensation reaction many 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 shortened production time for oligonucleic acid compounds. [Brief description of the drawings]

[0012] [Figure 1] Figure 1 illustrates a schematic diagram of a reactor used in a continuous reaction. F-1 to F-5 indicate solution containers, P-1 to P-5 indicate pumps, R-1 to R-4 indicate flow reactors, and S-1 to S-5 indicate supply channels. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] The present invention is a method for producing compound [C] by subjecting compound [A] having a hydroxyl group or a primary or secondary amino group to a condensation reaction with compound [B] having a substituent [1], characterized in that the reaction is carried out in the presence of at least one reaction promoter selected from the group consisting of quaternary ammonium salts, quaternary imidazolium salts, quaternary morpholinium salts, quaternary phosphonium salts, quaternary piperidinium salts, quaternary pyridinium salts, quaternary pyrrolidinium salts, and quaternary sulfonium salts.

[0014] (A) Regarding compound [A] The compound [A] that can be used in the present production method includes, for example, compounds having a hydroxyl group or a primary or secondary amino group.

[0015] One specific embodiment of compound [A] is a compound containing one to several nucleoside units in its molecule. Specifically, a compound containing 1 to 50 nucleoside units is suitable, a compound containing 1 to 30 nucleoside units is preferred, and a compound containing 1 to 25 nucleoside units is more preferred. Examples of the nucleoside units contained in the compound [A] include those represented by the following general formulae [4a] to [4d]: [ka] [In the formula, *teeth, (1) the position of the bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond; (2) The position of the bond to a hydrogen atom or (3) The following general formula [6]: [ka] (In the formula, * indicates the bond position with the residue of compound [A]; G is (1) silicon-substituted groups, (2) long chain alkyl-carbonyl, (3) Benzoyl substituted with 1 to 5 long chain alkyloxy and / or long chain alkenyloxy groups or (4) The following general formula [7]: [ka] (In the formula, * indicates the binding position with T; Z is (1) (soluble polymer soluble in organic solvent)-oxy, (2) (soluble polymer soluble in organic solvent)-amino, (3) long chain alkyloxy, (4) a solid support or (5) The following general formulas [8A] to [8N]: [ka] (In the formula, * indicates the bonding position with L; j represents an integer from 0 to 4; k represents an integer from 0 to 5; R 8a is a hydrogen atom or C 1-6 represents alkyl; R 8b are the same or different and each represents a long chain alkyl; R 8c are the same or different and are represented by the following general formulas [9A] to [9E]: [ka] (In the formula, * represents the bond position; and R 9 represents a long chain alkyl and / or a long chain alkenyl. (hereinafter referred to as "substituent [9A]", "substituent [9B]", "substituent [9C]", "substituent [9D]", and "substituent [9E]", respectively); R 8d are the same or different and represent a hydrogen atom, a halogen, a long-chain alkyl which may be substituted with 1 to 13 halogens, or a long-chain alkyloxy which may be substituted with 1 to 13 halogens; R 8e teeth, (1) Long-chain alkyl, (2) Long chain alkyl-carbonyl or (3) benzoyl substituted with 1 to 5 long chain alkyloxy and / or long chain alkenyloxy; and R 8f teeth, (1) Long-chain alkyl, (2) Long chain alkyl-carbonyl or (3) Long-chain alkenyl-carbonyl. (hereinafter referred to as "substituent [8A]", "substituent [8B]", "substituent [8C]", "substituent [8D]", "substituent [8E]", "substituent [8F]", "substituent [8G]", "substituent [8H]", "substituent [8I]", "substituent [8J]", "substituent [8K]", "substituent [8L]", "substituent [8M]", and "substituent [8N]", respectively); and L is the general formula

[10] : [ka] (In the formula, * represents the bond position to Z; ** indicates the bond position to the oxygen atom; and L 1 is optionally substituted C 2-10 Alkylene or optionally substituted C 6-10 It represents arylene. (hereinafter referred to as "substituent

[10] "). (hereinafter referred to as "substituent [7]"); and T is a single bond or the following general formula

[11] : [ka] (In the formula, X has the same meaning as above; W represents a lone pair, an oxygen atom, or a sulfur atom; * indicates the bond position to O; ** represents the bond position with G; and q represents an integer from 0 to 10. (hereinafter referred to as "substituent

[11] "), but when G is a silicon substituent, T is a single bond. (hereinafter referred to as "substituent [6]") represents the bonding position; **teeth, (1) the position of the bond between the phosphorus bond and the 3' oxygen atom or 3' nitrogen atom of the adjacent nucleoside unit; (2) The position of the bond to a hydrogen atom or (3) The bond position to the substituent [6]; d represents 0 or 1; B P represents an optionally protected nucleobase; R 4a is a hydrogen atom, a hydroxyl group substituted with a group that can be removed under neutral conditions, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkoxy-C 1-6 represents alkyl, halogen, nitro or cyano; R 4b1 and R 4b2 are the same or different and each represents a hydrogen atom or C 1-6 Represents alkyl or R 4b1 and R 4b2 together with the adjacent carbon atom to form a carbonyl; and J is an oxygen atom or NR 4b3 (R 4b3 is C 1-6 represents an alkyl group. (hereinafter referred to as “nucleoside unit [4a]”, “nucleoside unit [4b]”, “nucleoside unit [4c]”, and “nucleoside unit [4d]”, respectively).

[0016] Preferred embodiments of the nucleoside units [4a] to [4d] include, for example, the following general formulae [4a1] to [4d1]: [ka] [In the formula, d, B P ,J.R. 4a , R 4b1 and R 4b2 is as defined above; *teeth, (1) the position of the bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond; (2) The position of the bond to a hydrogen atom or (3) represents the bonding position to the substituent [6]; and **teeth, (1) the position of the bond between the phosphorus bond and the 3' oxygen atom or 3' nitrogen atom of the adjacent nucleoside unit; (2) The position of the bond to a hydrogen atom or (3) Represents the bonding position with the substituent [6]. (hereinafter referred to as “nucleoside unit [4a1]”, “nucleoside unit [4b1]”, “nucleoside unit [4c1]”, and “nucleoside unit [4d1]”, respectively).

[0017] When compound [A] contains a plurality of nucleoside units in its molecule, adjacent nucleoside units in the compound are preferably bonded to each other via a phosphorus bond. The phosphorus bonds between the nucleoside units constituting the compound [A] may be the same or different, and may be, for example, represented by the following general formula [5]: [ka] [In the formula, X has the same meaning as above; One of * and ** represents a bonding position between the 3' oxygen atom or the 3' nitrogen atom of a nucleoside unit, and the other represents a bonding position between the 5' oxygen atom of a nucleoside unit different from the nucleoside unit; and W represents a lone pair, an oxygen atom, or a sulfur atom. An example of such a bond is the phosphorus bond [5]. W preferably represents an oxygen atom or a sulfur atom, and more preferably represents an oxygen atom.

[0018] Representative examples of compound [A] are shown below.

[0019] (A-1) Compound [A] composed of one to several nucleoside units [4d] In the nucleoside unit represented by the general formula [4d], *but, (1) The position of the bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond, or (2) The position of the bond to a hydrogen atom; **but, (1) The position of the bond between the nitrogen atom at the 3' position of the adjacent nucleoside unit and the phosphorus bond, or (2) The bonding position with the substituent [6].

[0020] One embodiment of compound [A] is, for example, a compound in which the oxygen atom at the 5'-position of the 5'-terminal nucleoside unit is substituted with, for example, a substituent [6]. In this case, the phosphorus bonds between the nucleoside units constituting the compound [A] may be, for example, the same or different phosphorus bonds [5], where in the phosphorus bond represented by the general formula [5], one of * and ** represents the bonding position to the nitrogen atom at the 3' position of the nucleoside unit, and the other represents the bonding position to the oxygen atom at the 5' position of a nucleoside unit different from the nucleoside unit.

[0021] More specifically, the compound [A] may be represented by the following general formula [A-1]: [ka] [In the formula, B P , G, T, X and W are as defined above; and n represents an integer from 1 to 50. (hereinafter referred to as "compound [A-1]"). n is suitably an integer of 1 to 50, preferably an integer of 1 to 30, and more preferably an integer of 1 to 25. More specifically, the compound [A] may be represented by the following general formula [A-1-2]: Formula [A-1-2]: [ka] [In the formula, BP is an optionally protected nucleobase; Q 2 is H or a group removable under acidic conditions; W represents a lone electron pair, an oxygen atom, or a sulfur atom, preferably an oxygen atom or a sulfur atom, and more preferably an oxygen atom; X is di(C1-6 alkyl)amino; G is a member of the formula: [ka] [In the formula, * represents the bonding position with T; T is a single bond; n is 1 to 25. The compounds include:

[0022] (A-2) Compound [A] composed of one or more nucleoside units selected from the group consisting of nucleoside unit [4a], nucleoside unit [4b] and nucleoside unit [4c]. In the nucleoside units represented by the general formulae [4a], [4b] and [4c], *but, (1) The position of the bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond, or (2) the bonding position to the substituent [6]; **but, (1) The position of the bond between the oxygen atom at the 3' position of the adjacent nucleoside unit and the phosphorus bond, or (2) The bonding position with the hydrogen atom.

[0023] One embodiment of compound [A] is, for example, a compound in which the oxygen atom at the 3'-position of the 3'-terminal nucleoside unit is substituted with, for example, a substituent [6]. In this case, the phosphorus bond between each nucleoside unit constituting compound [A] is suitably, for example, phosphorus bond [5], where in the phosphorus bond represented by the general formula [5], one of * and ** represents the oxygen atom at the 3' position of a nucleoside unit, and the other represents the bond position between the nucleoside unit and the oxygen atom at the 5' position of a nucleoside unit different from the nucleoside unit. More specifically, the compound [A] may be represented by the following general formula [A-2]: [ka] [In the formula, n, B P , G, R 4a , T, X and W are as defined above.] (hereinafter referred to as "compound [A-2]").

[0024] Specific examples of the substituent [7] in the compound [A-1] and the compound [A-2] include the following substituents. [ka] [In the formula, * indicates the bond position with T.]

[0025] (B) Regarding compound [B] The compound [B] that can be used in this production method includes, for example, a compound having a substituent [1].

[0026] One specific embodiment of the compound [B] is, for example, a compound containing one to a plurality of nucleoside units in its molecule. More specifically, a compound containing 1 to 10 nucleoside units is suitable, a compound containing 1 to 7 nucleoside units is preferred, and a compound containing 1 to 5 nucleoside units is more preferred.

[0027] Examples of the nucleoside units contained in the compound [B] include those represented by the following general formulae [4e] to [4h]: [ka] [In the formula, d, B P ,J.R. 4a , R 4b1 and R 4b2 is as defined above; ***teeth, (1) the position of the bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond; (2) The bonding position of the substituent [1] or (3) represents the bond position to a group that can be removed under acidic conditions; ****teeth, (1) the position of the bond between the phosphorus bond and the 3' oxygen atom or 3' nitrogen atom of the adjacent nucleoside unit; (2) The bonding position of the substituent [1] or (3) Represents the bond position to a group that can be removed under acidic conditions.] (hereinafter referred to as “nucleoside unit [4e]”, “nucleoside unit [4f]”, “nucleoside unit [4g]”, and “nucleoside unit [4h]”, respectively).

[0028] Preferred embodiments of the nucleoside units [4e] to [4h] include, for example, the following general formulae [4e1] to [4h1]: [ka] [In the formula, d, B P ,J.R. 4a , R 4b1 and R 4b2 is as defined above; ***teeth, (1) the position of the bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond; (2) The bonding position of the substituent [1] or (3) represents the bond position to a group that can be removed under acidic conditions; and ****teeth, (1) the position of the bond between the phosphorus bond and the 3' oxygen atom or 3' nitrogen atom of the adjacent nucleoside unit; (2) The bonding position of the substituent [1] or (3) Represents the bond position to a group that can be removed under acidic conditions.] (hereinafter referred to as “nucleoside unit [4e1]”, “nucleoside unit [4f1]”, “nucleoside unit [4g1]”, and “nucleoside unit [4h1]”, respectively).

[0029] When compound [B] contains a plurality of nucleoside units in its molecule, adjacent nucleoside units in the compound are preferably bonded to each other via a phosphorus bond. In this case, the phosphorus bonds between the nucleoside units constituting the compound [B] may be, for example, the same or different phosphorus bonds [5], where one of * and ** in the phosphorus bond represented by the general formula [5] represents the 3' oxygen atom of the nucleoside unit, and the other represents the bond position between the nucleoside unit and the 5' oxygen atom of a nucleoside unit different from the nucleoside unit.

[0030] Representative examples of compound [B] are shown below.

[0031] (B-1) Compound [B] composed of one or more nucleoside units [4h] In the nucleoside unit represented by the general formula [4h], ***but, (1) The position of the bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond, or (2) a bond site to a group that can be removed under acidic conditions; ****but, (1) The position of the bond between the nitrogen atom at the 3' position of the adjacent nucleoside unit and the phosphorus bond, or (2) The bonding position with the substituent [1].

[0032] One embodiment of compound [B] is, for example, a compound in which the nitrogen atom at the 3'-position of the 3'-terminal nucleoside unit is substituted with a group that can be removed under acidic conditions. In this case, the phosphorus bonds between the nucleoside units constituting the compound [B] may be, for example, the same or different phosphorus bonds [5], where one of * and ** in the phosphorus bond represented by the general formula [5] represents the nitrogen atom at the 3' position of the nucleoside unit, and the other represents the bond position between the nucleoside unit and the oxygen atom at the 5' position of a nucleoside unit different from the nucleoside unit. The oxygen atom at the 5' position of the nucleoside unit on the 5' end of compound [B] has the following general formula [1A]: [ka] [In the formula, D, W and X are as defined above; and ** indicates the bonding position with the residue of compound [B].] It is suitable that the aryl group has a phosphorus atom-containing substituent represented by the following formula: More specifically, the compound [B] may be represented by the following general formula [B-1]: [ka] [In the formula, B P D, X and W are as defined above; p represents an integer from 1 to 10; and Q 1 represents a group that can be removed under acidic conditions.] (hereinafter referred to as "compound [B-1]"). In addition, p is suitably an integer of 1 to 10, preferably an integer of 1 to 7, and more preferably an integer of 1 to 5.

[0033] Specific examples of the compound [B-1] where p=1 include the compounds shown in Table 1 below.

[0034] [Table 1]

[0035] (B-2) A compound [B] composed of one or more nucleoside units selected from the group consisting of the nucleoside unit [4e], the nucleoside unit [4f] and the nucleoside unit [4g]. In the nucleoside units represented by the general formulae [4e], [4f] and [4g], ***but, (1) The position of the bond between the oxygen atom at the 3' position of the adjacent nucleoside unit and the phosphorus bond, or (2) the bonding position to the substituent [1]; ****but, (1) The position of the bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond, or (2) It is a bonding site with a group that can be removed under acidic conditions.

[0036] One embodiment of compound [B] is, for example, a compound in which the oxygen atom at the 5'-position of the 5'-terminal nucleoside unit is substituted with a group that can be removed under acidic conditions. In this case, the phosphorus bonds between the nucleoside units constituting the compound [B] may be, for example, the same or different phosphorus bonds [5], where one of the * and ** in the phosphorus bond represented by the general formula [5] represents the 3' oxygen atom of the nucleoside unit, and the other represents the bond position between the nucleoside unit and the 5' oxygen atom of a nucleoside unit different from the nucleoside unit. The oxygen atom at the 3' position of the nucleoside unit on the 3' end of compound [B] has the following general formula [1B]: [ka] [In the formula, D and X are as defined above; and ** indicates the bonding position with the residue of compound [B].] It is suitable that the aryl group has a phosphorus atom-containing substituent represented by the following formula: A more specific embodiment of the compound [B] is, for example, the compound represented by the following general formula [B-2]: [ka] [In the formula, p, B P , D, Q 1 , R 4a , X and W are as defined above.] (hereinafter referred to as "compound [B-2]").

[0037] Specific examples of the compound [B-2] where p=1 include the compounds shown in Table 2 below. In Table 2, DMTr represents dimethoxytrityl, and TBDMS represents tert-butyldimethylsilyl.

[0038] [Table 2]

[0039] (C) Regarding compound [C] An example of the compound [C] is a compound that can be produced by subjecting a compound [A] and a compound [B] to a condensation reaction.

[0040] Representative examples of compound [C] are shown below.

[0041] (C-1) Compound [C] composed of one or more nucleoside units [4d] and one or more nucleoside units [4h] Specific examples of the compound [C] include compounds represented by the following general formula [C-1]: [ka] [In the formula, n, p, B P , G, Q 1 , T, W and X are as defined above.] (hereinafter referred to as "compound [C-1]"). As described later, an example of the phosphorus bond newly formed in the method for producing compound [C-1] by reacting compound [A-1] with compound [B-1] is phosphorus bond [5]. In the phosphorus bond represented by the general formula [5], one of * and ** represents the nitrogen atom at the 3' position of a nucleoside unit, and the other represents the bond position between the nucleoside unit and the oxygen atom at the 5' position of a nucleoside unit different from the nucleoside unit.

[0042] (C-2) Compound [C] composed of one or more nucleoside units selected from the group consisting of nucleoside unit [4a], nucleoside unit [4b] and nucleoside unit [4c], and one or more nucleoside units selected from the group consisting of nucleoside unit [4e], nucleoside unit [4f] and nucleoside unit [4g].

[0043] One specific embodiment of the compound [C] is, for example, the compound represented by the following general formula [C-2]: [ka] [In the formula, n, p, B P , G, Q 1 , R 4a , T, W and X are as defined above.] (hereinafter referred to as "compound [C-2]"). As described later, examples of the phosphorus bond newly formed in the method for producing compound [C-2] by reacting compound [A-2] with compound [B-2] include the following general formula [5a]: [ka] [In the formula, X is as defined above; and One of * and ** represents the bonding position to the 3' oxygen atom of a nucleoside unit, and the other represents the bonding position to the 5' oxygen atom of a nucleoside unit different from the aforementioned nucleoside unit.] An example is a bond containing a phosphorus atom represented by the following formula (hereinafter referred to as "phosphorus bond [5a]").

[0044] By reacting the compound [C-2] with an oxidizing agent, the phosphorus atom on the phosphorus bond in the molecule is oxidized to the following general formula [D-2]: [ka] [In the formula, n, p, B P , G, Q 1 , R 4a , T, W and X are as defined above.] The compound represented by the formula (referred to as "compound [D-2]") can be converted into a compound represented by the formula (referred to as "compound [D-2]").

[0045] (D) Explanation of Terms Here, examples of "nucleic acid bases" include adenine, guanine, hypoxanthine, cytosine, thymine, uracil, and modified bases thereof. Examples of such modified bases include pseudouracil, 3-methyluracil, dihydrouracil, 5-alkylcytosine (e.g., 5-methylcytosine), 5-alkyluracil (e.g., 5-ethyluracil), 5-halouracil (5-bromouracil), 6-azapyrimidine, 6-alkylpyrimidine (6-methyluracil), 2-thiouracil, 4-thiouracil, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, 5'-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyluracil, 1-methyladenine, 1-methylhypoxanthine, 2,2-dimethylguanine, 3-methylcytosine, 2-methyladenine, 2-methylguanine, N 6 -Methyladenine, 7-methylguanine, 5-methoxyaminomethyl-2-thiouracil, 5-methylaminomethyluracil, 5-methylcarbonylmethyluracil, 5-methyloxyuracil, 5-methyl-2-thiouracil, 2-methylthio-N 6 -isopentenyladenine, uracil-5-oxyacetic acid, 2-thiocytosine, purine, 2,6-diaminopurine, 2-aminopurine, isoguanine, indole, imidazole, and xanthine, but are not limited thereto. P The amino group or hydroxyl group of the nucleic acid base of the formula (I) may be protected. As used herein, "optionally protected nucleobase" includes both unprotected and protected "nucleobases", such as adenine, guanine, hypoxanthine, cytosine, thymine, uracil, and the like, in which the amino and / or hydroxyl groups are unprotected or protected. The protecting group for the amino group is not particularly limited as long as it is used as a protecting group for nucleic acid, and specific examples thereof include benzoyl, 4-methoxybenzoyl, acetyl, propionyl, butyryl, isobutyryl, phenylacetyl, phenoxyacetyl, 4-tert-butylphenoxyacetyl, 4-isopropylphenoxyacetyl, and (dimethylamino)methylene. The protecting group for the amino group is preferably benzoyl, acetyl, phenylacetyl, or 4-tert-butylphenoxyacetyl. Examples of the protective group for the hydroxyl group include 2-cyanoethyl, 4-nitrophenethyl, phenylsulfonylethyl, methylsulfonylethyl, trimethylsilylethyl, phenyl which may be 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). Examples of the protective group for the hydroxyl group include 2-cyanoethyl, 4-nitrophenethyl, and 4-(tert-butylcarboxy)benzyl. The protective group for the 6-position hydroxyl group of guanine is preferably 2-cyanoethyl. In one embodiment, protected nucleobases include, for example, those shown below: [ka] [In the formula, Pg represents a protecting group]. A more specific embodiment of the protected nucleic acid base is adenine (A) in which the amino group is protected with benzoyl. Bz), cytosine with the amino group protected by benzoyl (C Bz ), guanine (G) whose hydroxyl group is protected with 2-cyanoethyl and whose amino group is protected with phenoxyacetyl; CE,Pac ), but are not limited to these. The "long-chain alkyl" refers to, for example, a linear or branched alkyl having 10 to 300 carbon atoms, preferably a linear or branched alkyl having 10 to 100 carbon atoms, and more preferably a linear or branched alkyl having 10 to 30 carbon atoms. The "long chain alkyl" portion of "long chain alkyl-carbonyl" and "long chain alkyloxy" can be the same as the "long chain alkyl" defined above. "Long-chain alkenyl" refers to, for example, linear or branched alkenyl having 10 to 300 carbon atoms, preferably linear or branched alkenyl having 10 to 100 carbon atoms, and more preferably 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" defined above. Examples of the "halogen" include a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom. Examples of the "5- to 6-membered saturated cyclic amino" include a 5- to 6-membered saturated cyclic amino group having 1 or 2 N, which may have 1 O or S 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. "C 1-6 "Alkyl" refers to linear or branched alkyl having 1 to 6 carbon atoms, and specific examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl. "C 1-6"Alkoxy" refers to straight-chain or branched-chain alkoxy having 1 to 6 carbon atoms, and specific examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentyloxy, and n-hexyloxy. "C 1-6 Alkoxy-C 1-6 "C" in "Alkyl" 1-6 The "alkoxy" portion is 1-6 The same as "alkoxy" can be mentioned. "Ji (C 1-6 Alkyl)amino", mono(under basic conditions Detachment Amino-C substituted with possible groups 1-6 Alkyl)amino, di(basic conditions Detachment Amino-C substituted with possible groups 1-6 Alkyl)amino, mono(C 1-6 Alkyl)amino-C 1-6 Alkyl, di(C 1-6 Alkyl)amino-C 1-6 Alkyl, tri(C 1-6 Alkyl)ammonio-C 1-6 Alkyl, mono(C 1-6 Alkyl)amino, di(C 1-6 Alkyl)amino, tri(C 1-6 Alkyl)ammonio, mono(amino-C 1-6 Alkyl)amino and di(amino-C 1-6 (alkyl)amino "C 1-6 The "C alkyl" portion is 1-6 The same can be mentioned as "alkyl". "C 2-10 "Alkylene" is a divalent group generated by removing one hydrogen atom bonded to a different constituent carbon atom from a linear or branched alkyl group 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 halogens at any substitutable position. L 1As the "alkylene" in the above, ethylene is particularly preferred. "C 6-10 "Arylene" is a divalent group generated 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 an "arylene" may be substituted with 1 to 6 halogens at any substitutable position. L 1 As the "arylene" in the above, phenylene is particularly preferable. "1,1,3,3-tetra(C 1-6 alkyl) guanidyl", "C 1-6 Alkoxy-C 1-6 Alkyl, Di(C 1-6 alkyl)amino", "di(C 1-6 Alkyl)amino-C 1-6 Alkyl, Tri(C 1-6 alkyl)ammonio," "tri(C 1-6 Alkyl)ammonio-C 1-6 Alkyl," "mono (C 1-6 "Mono (C alkyl) amino" or "mono (C 1-6 Alkyl)amino-C 1-6 Alkyl, mono(amino-C substituted with a group that can be removed under basic conditions 1-6 "Di(alkyl)amino" and "Di(amino-C substituted with a group that can be removed under basic conditions 1-6 (alkyl)amino" 1-6 The "C alkyl" portion is 1-6 The same can be mentioned as "alkyl". "Under acidic conditions Detachment Examples of the "possible groups" include trityl, monomethoxytrityl, tert-butyldimethylsilyl, and dimethoxytrityl. "Under basic conditions Detachment An example of a "possible group" is trifluoroacetyl. "Under neutral conditions DetachmentExamples of the "removable group" include groups that can be removed by the action of tetrabutylammonium fluoride or hydrogen trifluoride-triethylamine salt, such as 2-cyanoethoxymethoxy, 2-cyanoethoxy-2-ethoxy, and tert-butyldimethylsilyl. Examples of the "silicon substituent" include triphenylsilyl, diisopropylphenylsilyl, tert-butyldimethylsilyl, and tert-butyldiphenylsilyl. "Aryl" can be, for example, phenyl. Examples of "heteroaryl" include pyridyl, pyrimidyl, pyridazyl, pyrazinyl, thienyl, and furanyl. As the "solid phase support", generally, any support that can be used in the solid phase synthesis of nucleic acids, peptides, peptide nucleic acids, sugars, and the like can be used without any particular problems. Examples of such support include controlled pore glass (CPG), oxalyl-controlled pore glass (see, for example, Nucleic Acids Research, Vol. 19, 1527 (1991)), TentaGel support-amino polyethylene glycol derivatized support (see, for example, Tetrahedron Letters, Vol. 34, 3373 (1993)), Poros-polystyrene / divinylbenzene copolymer, polystyrene resin, and polyacrylamide resin. Examples of the "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 polystyrene derivatives that are not crosslinked with divinylbenzene and have a spacer such as polyethylene glycol (TentaGel series, ArgoGel series). Examples of the "polyethylene glycol derivative" include derivatives having a substituent in polyethylene glycol with a molecular weight of 100 to 40,000 (SUNBRIGHT (registered trademark) series).

[0046] (E) Method for producing compound [C] For example, compound [C] can be produced by subjecting compound [A] having a hydroxyl group or a primary or secondary amino group to a condensation reaction with compound [B] having a substituent [1]. As described in the Examples and Test Examples below, in the production of compound [C], phosphorus bonds can be efficiently formed in the presence of a reaction promoter.

[0047] The solvent that can be used in the present production method 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. Examples of solvents that can be used in the present production method include aromatic solvents such as benzene, toluene, xylene, and mesitylene, ester solvents such as ethyl acetate and isopropyl acetate, and aliphatic solvents such as hexane, pentane, heptane, octane, nonane, and cyclohexane. These solvents may be used in combination. In this production method, a base may be used as necessary. Examples of the "base" that can be used in this production method include diisopropylamine, N,N-diisopropylethylamine, triethylamine, N-ethylmorpholine, and 2,6-lutidine. The amount of base that can be used in the present production method is, for example, appropriately within the range of 1 to 100 times, preferably 1 to 10 times, and more preferably 1 to 5 times, the molar ratio of the base to 1 mole of compound [A]. In this production method, a reaction promoter is used. The "reaction promoter" that can be used in this production method can be, for example, at least one selected from the group consisting of quaternary ammonium salts, quaternary imidazolium salts, quaternary morpholinium salts, quaternary phosphonium salts, quaternary piperidinium salts, quaternary pyridinium salts, quaternary pyrrolidinium salts, and quaternary sulfonium salts, and preferably at least one selected from the group consisting of quaternary ammonium salts, quaternary imidazolium salts, quaternary phosphonium salts, quaternary pyridinium salts, and quaternary pyrrolidinium salts. More preferably, a salt containing a quaternary nitrogen cation selected from the group consisting of quaternary ammonium salts, quaternary imidazolium salts, quaternary pyridinium salts, and quaternary pyrrolidinium salts, and even more preferably, at least one selected from the group consisting of quaternary ammonium salts, quaternary imidazolium salts, and quaternary pyrrolidinium salts can be used. Examples of quaternary ammonium salts that can be used in the present production method include amyltriethylammonium bis(trifluoromethanesulfonyl)imide, butyltrimethylammonium bis(trifluoromethanesulfonyl)imide, benzyl(ethyl)dimethylammonium bis(trifluoromethanesulfonyl)imide, cyclohexyltrimethylammonium bis(trifluoromethanesulfonyl)imide, diethyl(methyl)propylammonium bis(fluorosulfonyl)imide, diethyl(2-methoxyethyl)methylammonium bis(fluorosulfonyl)imide, ethyl(2-methoxyethyl)dimethylammonium bis(fluorosulfonyl)imide, ethyl(2-methoxyethyl)dimethylammonium bis(trifluoromethanesulfonyl)imide, ethyl(3-methoxypropyl)dimethylammonium bis(trifluoromethanesulfonyl)imide, ethyl(dimethyl)(2-phenylethyl)ammonium bis(trifluoromethanesulfonyl)imide, methyltri-n-octylammonium bis(trifluoromethanesulfonyl)imide, tetrafluoroethylene ... tetrabutylammonium bromide, tetrabutylammonium chloride, tetrabutylammonium iodide, tetrabutylammonium tetrafluoroborate, tetrahexylammonium iodide, tetraamylammonium iodide, tetra-n-octylammonium iodide, tetrabutylammonium hexafluorophosphate, tetraheptylammonium iodide iodide, tetraamylammonium bromide, tetraamylammonium chloride, tetrabutylammonium trifluoromethanesulfonate, tetrahexylammonium bromide, tetraheptylammonium bromide, tetra-n-octylammonium bromide, tetrapropylammonium chloride, tributylmethylammonium bis(trifluoromethanesulfonyl)imide, tetrabutylammonium acetate, trimethylpropylammonium bis(trifluoromethanesulfonyl)imide, tributyl(methyl)ammonium dicyanamide, tetrabutylammonium p-toluenesulfonate, tributylmethylammonium iodide, and the like. The quaternary ammonium salt that can be used in this production method is, for example, tetra C 1-18 Alkyl ammonium salts (e.g., tetra C 1-18 Alkyl ammonium chloride), Tri C 1-18 Alkyl(hydroxy C 1-18 alkyl) ammonium salts, etc. (wherein 1-18 alkyl may be the same or different), preferably tetramethylammonium chloride, tetraethylammonium chloride, tetrapropylammonium chloride, tetrabutylammonium chloride, tetrabutylammonium bromide, dodecyltrimethylammonium chloride, choline chloride, N,N,N-trimethylbutane-1-amino chloride. Examples of quaternary imidazolium salts that can be used in this production method include 1-allyl-3-methylimidazolium chloride, 1-allyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, 1-butyl-3-methylimidazolium bromide, 1-butyl-3-methylimidazolium chloride, 1-butyl-3-methylimidazolium tetrafluoroborate, 1-butyl-3-methylimidazolium hexafluorophosphate, 1-butyl-3-methylimidazolium trifluoromethanesulfonate, and 1-butyl-2,3-dimethylimidazolium tetrafluoroborate. Midazolium chloride, 1-butyl-2,3-dimethylimidazolium hexafluorophosphate, 1-butyl-2,3-dimethylimidazolium tetrafluoroborate, 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, 1-butyl-3-methylimidazolium tetrachloroferrate, 1-butyl-3-methylimidazolium iodide, 1-butyl-2,3-dimethylimidazolium bis(trifluoromethanesulfonyl)imide, 1-butyl-3-methylimidazolium methanesulfonate, 1-butyl -3-Methylimidazolium trifluoro(trifluoromethyl)borate, 1-butyl-3-methylimidazolium tribromide, 1-butyl-3-methylimidazolium thiocyanate, 1-butyl-2,3-dimethylimidazolium trifluoromethanesulfonate, 3,3'-(butane-1,4-diyl)bis(1-vinyl-3-imidazolium)bis(trifluoromethanesulfonyl)imide, 1-butyl-3-methylimidazolium dicyanamide, 1-butyl-3-methylimidazolium tricyanomethan ...1-butyl-2,3-dimethylimidazolium trifluoromethanesulfonate, 1-butyl-2,3-dimethylimidazolium trifluoromethanesulfonate, 1-butyl-2,3-dimethylimidazolium trifluoromethanesulfonate, 1-butyl-2,3-dimethylimidazolium trifluoromethanesulfonate, 1-butyl-2,3-dimethylimidazolium trifluoromethanesulfonate, 1-butyl-2,3-dimethylimidazolium trifluoromethanesulfonate, 1-butyl-2,3-dimethylimidazolium trifluoromethanesulfonate, 1-butyl-2,3-dimethylimidazolium trifluoromethanesulfonate, 1-butyl-2 1-Butyl-3-methylimidazolium trifluoroacetate, 1-Butyl-3-methylimidazolium methylsulfate, 1-Benzyl-3-methylimidazolium chloride, 1-Butyl-3-methylimidazolium hydrogen sulfate, 1-Butyl-3-methylimidazolium dibutyl phosphate, 1-Butyl-3-methylimidazolium hexafluoroantimonate, 1-Benzyl-3-methylimidazolium tetrafluoroborate, 1-Benzyl-3-methylimidazolium hexafluorophosphate, 1,3-Dimethylimidazolium dimethyl phosphate, 1,3-Dimethylimidazolium chloride, 1,2-dimethyl-3-propylimidazolium iodide, 2,3-dimethyl-1-propylimidazolium bis(trifluoromethanesulfonyl)imide, 1-decyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, 1,3-dimethylimidazolium iodide, 1,3-dimethylimidazolium methylsulfate, 1,3-dimethylimidazolium bis(trifluoromethanesulfonyl)imide, 1-decyl-3-methylimidazolium bromide, 1-decyl-3-methylimidazolium chloride, 1-dodecyl-3-methylimidazolium bromide, 1-decyl-3-methylimidazolium tetrafluoroborate, 1-dodecyl-3-methylimidazolium bis(trifluoro methanesulfonyl)imide, 1-ethyl-3-methylimidazolium chloride, 1-ethyl-3-methylimidazolium hexafluorophosphate, 1-ethyl-3-methylimidazolium trifluoromethanesulfonate, 1-ethyl-3-methylimidazolium tetrafluoroborate, 1-ethyl-3-methylimidazolium bromide, 1-ethyl-3-methylimidazolium iodide, 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, 1-ethyl-3-methylimidazolium ethyl sulfate, 1-ethyl-3-methylimidazolium p-toluenesulfonate, 1-ethyl-3-methylimidazolium dicyanamide, 1-ethyl-3-methylimidazolium tetrachloroferrate, 1-ethyl-2,3-Dimethylimidazolium bis(trifluoromethanesulfonyl)imide, 1-ethyl-3-methylimidazolium hydrogen sulfate, 1-ethyl-3-methylimidazolium methanesulfonate, 1-ethyl-3-methylimidazolium nitrate, 1-ethyl-3-methylimidazolium thiocyanate, 1-ethyl-3-methylimidazolium trifluoro(trifluoromethyl)borate, 1-ethyl-3-methylimidazolium acetate, 3-ethyl-1-vinylimidazolium bis(trifluoromethanesulfonyl)imide, 1-ethyl-3-methylimidazolium tricyanomethanide , 1-ethyl-3-methylimidazolium trifluoroacetate, 1-ethyl-3-methylimidazolium methylsulfate, 1-ethyl-3-methylimidazolium diethylphosphate, 1-hexyl-3-methylimidazolium chloride, 1-hexyl-3-methylimidazolium hexafluorophosphate, 1-hexyl-3-methylimidazolium tetrafluoroborate, 1-hexyl-3-methylimidazolium trifluoromethanesulfonate, 1-hexyl-3-methylimidazolium bromide, 1-(2-hydroxyethyl)-3-methylimidazolium chloride, 1-hexyl-2,3-Dimethylimidazolium iodide, 1-hexyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, 1-(2-hydroxyethyl)-3-methylimidazolium tetrafluoroborate, 1-hexyl-3-methylimidazolium iodide, 1-(2-hydroxyethyl)-3-methylimidazolium bis(trifluoromethanesulfonyl)imide, 1-methyl-3-propylimidazolium iodide, 1-methyl-3-N-octylimidazolium bromide, 1-methyl-3-N-octylimidazolium chloride, 1-methyl-3-N-octylimidazolium hexafluorophosphate, 1-methyl-3-N-octylimidazolium trifluoromethanesulfonate, 1-methyl-3-N-octylimidazolium tetrafluoroborate ester, 1-methyl-3-propylimidazolium bromide, 1-methyl-3-propylimidazolium chloride, 1-methyl-3-propylimidazolium tetrafluoroborate, 1-methyl-3-pentylimidazolium bromide, 1-methyl-3-N-octylimidazolium bis(trifluoromethanesulfonyl)imide, 1-methyl-3-propylimidazolium bis(trifluoromethanesulfonyl)imide, 1-methyl-3-(4-sulfobutyl)imidazolium bis(trifluoromethanesulfonyl)imide, 1-methyl-3-(4-sulfobutyl)imidazolium hydrogen sulfate, 1-methylimidazole, preferably 1-ethyl-3-methylimidazolium chloride, 1-methyl-3-N-octylimidazolium chloride. The quaternary imidazolium salt that can be used in this production method is, for example, 1-C 1-18 Alkyl-3-C 1-18 Alkyl imidazolium chloride, 1-C 1-18 Alkyl-3-NC 1-18 The alkylimidazolium chloride is preferably 1-ethyl-3-methylimidazolium chloride or 1-methyl-3-N-octylimidazolium chloride. The quaternary phosphonium salt that can be used in this production method is, for example, tetra C 1-18 Alkylphosphonium salts, TriC 1-18Alkyl(hydroxy C 1-18 alkyl)phosphonium salts, 1-18 The alkyl groups may be the same or different, preferably trihexyltetradecylphosphonium chloride. The quaternary pyridinium salt that can be used in this production method is, for example, 1-C 1-18 The alkylpyridinium salt is preferably 1-ethylpyridinium bromide, 1-ethylpyridinium chloride, or 1-butylpyridinium chloride. Examples of quaternary pyrrolidinium salts that can be used in this production method include 1-allyl-1-methylpyrrolidinium bis(trifluoromethanesulfonyl)imide, 1-butyl-1-methylpyrrolidinium bis(trifluoromethanesulfonyl)imide, 1-butyl-1-methylpyrrolidinium chloride, 1-butyl-1-methylpyrrolidinium bromide, 1-butyl-1-methylpyrrolidinium bis(fluorosulfonyl)imide, 1-butyl-1-methylpyrrolidinium dicyanamide, 1-butyl-1-methylpyrrolidinium trifluoromethanesulfonate, 1-butyl-1-methylpyrrolidinium hexafluorophosphate, 1-ethyl 1-methylpyrrolidinium tetrafluoroborate, 1-ethyl-1-methylpyrrolidinium bromide, 1-methyl-1-propylpyrrolidinium bis(trifluoromethanesulfonyl)imide, 1-methyl-1-propylpyrrolidinium bis(fluorosulfonyl)imide, 1-(2-methoxyethyl)-1-methylpyrrolidinium bis(fluorosulfonyl)imide, 1-methyl-1-N-octylpyrrolidinium bis(trifluoromethanesulfonyl)imide, 1-methyl-1-pentylpyrrolidinium bis(trifluoromethanesulfonyl)imide, and preferably 1-butyl-1-methylpyrrolidinium chloride. The quaternary pyrrolidinium salt that can be used in this production method is, for example, 1-C 1-18 Alkyl-1-C 1-18 Alkylpyrrolidinium chloride (where C 1-18 The alkyl groups may be the same or different, and preferably 1-butyl-1-methylpyrrolidinium chloride. Examples of reaction accelerators that can be used in this production method include: Tetramethylammonium chloride, Tetraethylammonium chloride, Tetrapropylammonium chloride, Tetrabutylammonium chloride, tetrabutylammonium bromide, dodecyltrimethylammonium chloride, Trioctylmethylammonium chloride, N,N,N-trimethylbutane-1-amino chloride, 1-ethylpyridinium bromide, 1-ethylpyridinium chloride, 1-butylpyridinium chloride, 1-ethyl-3-methylimidazolium chloride, 1-Methyl-3-N-octylimidazolium chloride, 1-Butyl-1-methylpyrrolidinium chloride, and Trihexyltetradecylphosphonium chloride At least one selected from the group consisting of: In one embodiment, a preferred reaction promoter that can be used in the present production method is Tetra C 1-18 Alkyl ammonium chloride, 1-C 1-18 Alkyl-1-C 1-18 Alkylpyrrolidinium chloride, 1-C 1-18 Alkyl-3-C 1-18 Examples of the alkylimidazolium chloride include alkylimidazolium chloride, and preferably at least one selected from the group consisting of tetrabutylammonium chloride, 1-butyl-1-methylpyrrolidinium chloride, and 1-methyl-3-N-octylimidazolium chloride. The amount of the reaction accelerator that can be used in the present production method is, in terms of molar ratio, within the range of 1 to 100 times, preferably within the range of 1 to 50 times, and more preferably within the range of 1.5 to 20 times, per 1 mole of compound [A]. The reaction temperature is suitably within the range of -78°C to 130°C, for example, preferably within the range of -40°C to 100°C, and more preferably within the range of 0°C to 80°C. The reaction time varies depending on the type of compound [A], type of compound [B], type of reaction solvent, type of base, and reaction temperature used, but is, for example, appropriately within the range of 1 minute to 300 minutes, and preferably within the range of 5 minutes to 120 minutes.

[0048] When compound [C] which is an oligonucleic acid compound can be produced, the production method can be applied as either a batch method or a flow method. Furthermore, the present production method can also be applied to known methods for producing oligonucleic acid compounds, such as a solid-phase method, a liquid-phase method, and a liquid-phase method using hydrophobic group-bonded nucleosides or pseudo-solid-phase protected nucleosides.

[0049] When the oligonucleic acid compound, compound [C], can be produced using a solid phase method, compound [A] can be supported on a solid phase carrier at the 3' oxygen atom of the nucleoside unit at the 3' end or at the 5' oxygen atom of the nucleoside unit at the 5' end.

[0050] When an oligonucleic acid compound can be produced using a liquid phase method, the 3'-position oxygen atom of the nucleoside unit on the 3'-terminal side of compound [A] or the 5'-position oxygen atom of the nucleoside unit on the 5'-terminal side of compound [A] can be supported on a soluble polymer that dissolves in an organic solvent.

[0051] When an oligonucleic acid compound can be produced by a liquid phase method using a hydrophobic group-bound nucleoside or a pseudo solid phase-protected nucleoside, for example, a nucleoside unit having a hydrophobic group bound to the 3' oxygen atom of the 3'-terminal nucleoside unit of compound [A] or a nucleoside unit having a 5' oxygen atom of the 5'-terminal nucleoside unit of compound [A] can be used (see, for example, JP 2010-275254 A and WO 2012 / 157723 A).

[0052] The compound [C-1] and the compound [C-2] will be described in detail below as examples.

[0053] (E-1) Method for producing compound [C-1] [ka] [In the formula, n, p, B P , D, G, Q 1 , T, W and X are as defined above.]

[0054] Compound [C-1] can be produced by subjecting compound [A-1] to a condensation reaction with compound [B-1]. The solvent that can be used in the present production method is not particularly limited as long as it is a solvent that is commonly used in the technical field, and the solvent may be used alone or in a mixture of two or more solvents. For example, aromatic solvents such as benzene, toluene, xylene, mesitylene, etc.; ester solvents such as ethyl acetate, isopropyl acetate, etc.; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, cyclohexane, etc. Two or more of these solvents may be used in a mixture. Solvents that can be used in the present production method include, for example, polar solvents and halogenated solvents. Examples of polar solvents that can be used in the present production method include dimethylacetamide, dimethylsulfoxide, dimethylformamide, sulfolane, N-methylpiperidone, 1,3-dimethyl-2-imidazolidinone, N,N'-dimethylpropyleneurea, and mixtures thereof. Among these, dimethylacetamide, dimethylsulfoxide, N-methylpiperidone, 1,3-dimethyl-2-imidazolidinone, and N,N'-dimethylpropyleneurea are preferred. Examples of halogen-based solvents that can be used in the present production method include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, and mixtures thereof. Among these, chloroform, dichloromethane, 1,1-dichloroethane, and 1,2-dichloroethane are preferred. The ratio of the polar solvent in the mixed solvent of the polar solvent and the halogen-based solvent that can be used in the present production method is 1.0% by weight, preferably 2.0% by weight, more preferably 3.0% by weight, and even more preferably 5.0% by weight. The upper limit is 90% by weight, preferably 75% by weight, more preferably 50% by weight, even more preferably 40% by weight, and particularly preferably 30% by weight. Furthermore, the upper and lower limits can be used in appropriate combination, and for example, the range of 1% to 90% is appropriate, the range of 5% to 75% is preferable, and the range of 5% to 50% is particularly preferable. In this production method, a base may be used as necessary. Examples of the "base" that can be used in this production method include diisopropylamine, N,N-diisopropylethylamine, triethylamine, N-ethylmorpholine, and 2,6-lutidine. The amount of base that can be used in the present production method is, for example, appropriately within the range of 1 to 100 times, preferably 1 to 10 times, and more preferably 1 to 5 times, the molar ratio of the base to 1 mole of compound [A]. In the present production method, an additive may be used as necessary. Preferred examples of the "additive" that can be used in the present production method include LiBr, LiCl, LiI, and NaI. The amount of additives that can be used in the present production method is, for example, appropriately within the range of 0.2 to 6.0 times the molar amount per 1 mole of compound [A], preferably within the range of 0.4 to 3.0 times the molar amount, and more preferably within the range of 1.0 to 2.5 times the molar amount. The reaction temperature is suitably within the range of -78°C to 130°C, for example, preferably within the range of -40°C to 100°C, and more preferably within the range of 0°C to 80°C. The reaction time varies depending on the type of compound [A], type of compound [B], type of reaction solvent, type of base, and reaction temperature used, but is, for example, appropriately within the range of 1 minute to 300 minutes, and preferably within the range of 5 minutes to 120 minutes.

[0055] When compound [A-1] has a solid phase carrier in its molecule, i.e., when G in compound [A-1] is a substituent [7] and Z is a solid phase carrier, this condensation reaction can be carried out, for example, by (1) packing compound [A-1] into an appropriate column and eluting a reaction solution containing compound [B-1], or (2) shaking or stirring a reaction solution containing compound [A-1] and compound [B-1] in a reaction vessel equipped with a filter.

[0056] In the compound [A-1], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (4) a substituent [7] (excluding the case where Z is a solid phase support), for example, the present condensation reaction can be carried out by (1) stirring compound [A-1] and compound [B-1] in a reaction solvent in a suitable reaction vessel, or (2) supplying a solution containing compound [A-1] and a solution containing compound [B-1] independently through a supply flow path into a flow reactor or a reaction flow path, and mixing the solutions in the flow reactor or the like.

[0057] Here, the term "supply flow path" refers to a flow path for continuously supplying a solution, the term "reaction flow path" refers to a flow path through which a solution can be reacted while flowing, and the term "flow reactor" refers to a reactor in which the introduction of a solution, reaction, and recovery of the product are carried out simultaneously, allowing uninterrupted operation. Examples of a means for supplying a solution containing compound [A-1] and a solution containing compound [B-1] to the supply flow path include pumps typically used in this field for supplying liquids, specifically, for example, a syringe pump, a plunger pump, a diaphragm pump, and a gear pump. Examples of the flow reactor include a microreactor and an in-line mixer such as a static mixer. An example of a means for introducing the solution containing the compound [A-1] and the solution containing the compound [B-1] from the supply channel to the reaction channel is a multi-stage collision type micromixer. Examples of materials for the supply flow path and the reaction flow path include tubes made of synthetic resins selected from the group consisting of fluorine-based resins such as perfluoroalkoxyalkane (PFA), vinyl chloride-based resins, polyamide-based resins, and aromatic polyether ketone-based resins, and tubes made of metals selected from the group consisting of stainless steel, copper and its alloys, and titanium and its alloys. The inner diameter of the supply channel and the reaction channel may be appropriately selected, for example, from a range of usually 0.1 mm to 1.0 mm, and is preferably selected, for example, from a range of 0.2 mm to 1.0 mm.

[0058] (E-2) Method for producing compound [C-2] [ka] [In the formula, n, p, B P , D, G, Q 1 , R 4a , T, W and X are as defined above.]

[0059] Compound [C-2] can be produced by subjecting compound [A-2] to a condensation reaction with compound [B-2]. In this production method, a base may be used as necessary. Examples of the "base" that can be used in this production method include diisopropylamine, N,N-diisopropylethylamine, triethylamine, N-ethylmorpholine, and 2,6-lutidine.

[0060] When compound [A-2] has a solid phase carrier in its molecule, i.e., when G in compound [A-2] is a substituent [7] and Z is a solid phase carrier, this condensation reaction can be carried out, for example, by (1) packing compound [A-1] into an appropriate column and eluting a reaction solution containing compound [B-2], or (2) shaking or stirring a reaction solution containing compound [A-2] and compound [B-2] in a reaction vessel equipped with a filter.

[0061] In the compound [A-2], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (4) a substituent [7] (excluding the case where Z is a solid phase support), for example, the present condensation reaction can be carried out by stirring compound [A-2] and compound [B-2] in a reaction solvent in a suitable reaction vessel, or by supplying a solution containing compound [A-2] and a solution containing compound [B-2] independently through a supply flow path into a flow reactor or a reaction flow path, and mixing the solutions in the flow reactor or the like. Furthermore, after the condensation reaction, (1) the reaction mixture can be purified using a column to obtain compound [C-2], or (2) an appropriate solvent can be added to the reaction mixture to obtain a precipitate, which can be collected by filtration and washed with an appropriate solvent to obtain compound [C-2].

[0062] Compound [C-2] is a compound in which each nucleoside unit constituting the compound is nucleoside unit [4a] and nucleoside unit [4e]. However, even if the compound is one in which all or a part of nucleoside unit [4a] or nucleoside unit [4e] is replaced with nucleoside unit [4b], nucleoside unit [4c], nucleoside unit [4f] or nucleoside unit [4g], it can be produced in the same manner as above.

[0063] (F) Purification method for compound [C] When compound [C] has a substituent in its molecule that exhibits extremely high lipophilicity, it can be easily isolated and purified by crystallization or extraction alone, without the need for complicated procedures such as column purification. Examples of such compounds include compound [C-1] and compound [C-2] in which G is (1) a long-chain alkyl-carbonyl, (2) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (3) a substituent [7] (excluding the case where Z is a solid phase support). On the other hand, when compound [C] has a solid phase carrier in its molecule, it can be purified, for example, by packing compound [C] in an appropriate column and washing compound [C] with an appropriate solvent to remove undesired substances. Examples of such compounds include compounds [C-1] and [C-2] in which G is a substituent [7] and Z is a solid phase carrier. In addition, in the compounds [C-1] and [C-2], when G is a silicon substituent, the target compound can be isolated and purified by performing operations such as column purification using an appropriate solvent.

[0064] (G) Q in the molecule of compound [C] 1 Method of detachment When compound [C] is a compound containing two or more nucleoside units, the molecule may have a hydroxyl group or a primary or secondary amino group protected by a group that can be removed under acidic conditions. In such cases, the groups that can be removed under acidic conditions in the molecule are selectively Detachment By carrying out the condensation reaction described in "(E) Method for producing compound [C]" above on the new compound produced by the above process, compound [C] having one more nucleoside unit can be produced.

[0065] The compound [C-1] and the compound [C-2] will be described in detail below as examples.

[0066] (G-1) Q in the molecule of compound [C-1] 1 Method of detachment The compound [C-1] is reacted with an acid to obtain a compound [C-2] having a nitrogen atom at the 3' position of the nucleoside unit at the 3' end of the compound [C-2], which is substituted with Q. 1 Q in the molecule of compound [C-1] can be eliminated. 1 By removing the above, a compound represented by the following general formula [E-1] (hereinafter referred to as "compound [E-1]") can be produced.

[0067] [ka] [In the formula, n, p, B P , G, Q 1 , T, W and X are as defined above.]

[0068] In one embodiment, the Q substituted at the 5'-position oxygen atom of the 5'-terminal nucleoside of the compound [C-1-1] 1 Before the elimination, the phosphorus atom on the phosphorus bond formed in the condensation reaction can first be oxidized from trivalent to pentavalent using an oxidizing agent, thereby converting it into a compound represented by general formula [C-1] (hereinafter referred to as "compound [C-1]"). [ka]

[0069] When G is a substituent [7] and Z is a solid phase support in compound [C-1], the elimination reaction can be carried out, for example, by (1) packing compound [C-1] into an appropriate column and eluting it with a solution containing an acid, or (2) shaking or stirring a solution containing compound [C-1] and an acid in a reaction vessel equipped with a filter. The solvent that can be used in this elimination reaction is not particularly limited as long as it is a solvent that is commonly used in the technical field, and the solvent may be used alone or in a mixture of two or more solvents. For example, aromatic solvents such as benzene, toluene, xylene, mesitylene, etc.; ester solvents such as ethyl acetate, isopropyl acetate, etc.; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, cyclohexane, etc. Two or more of these solvents may be used in a mixture. Solvents that can be used in this elimination reaction include, for example, polar solvents and halogenated solvents. The solvent that can be used in this elimination reaction is not particularly limited, but for example, a mixed solvent of a polar solvent and a halogen-based solvent can be used. The ratio of the polar solvent in the mixed solvent of the polar solvent and the halogen-based solvent is 1.0% by weight, preferably 2.0% by weight, more preferably 3.0% by weight, and even more preferably 5.0% by weight. The upper limit is 90% by weight, preferably 75% by weight, more preferably 50% by weight, even more preferably 40% by weight, and particularly preferably 30% by weight. Furthermore, the upper and lower limits can be used in appropriate combination, and for example, the range of 1% to 50% is appropriate, the range of 1% to 40% is preferable, and the range of 1% to 30% is particularly preferable. Examples of the "acid" that can be used in this elimination reaction 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. The amount of the acid that can be used in this elimination reaction is suitably within the range of 1 to 500 times, and preferably 2 to 200 times, the molar ratio of which is based on 1 mole of the compound [C-1]. The acid that can be used in this elimination reaction is suitably diluted with a suitable solvent to a concentration within the range of, for example, 5% to 80%, and preferably diluted to a concentration within the range of 5% to 50%. The solvent for dissolving the acid that can be used in this elimination reaction is not particularly limited, but examples thereof include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, and mixed solvents thereof. In this elimination reaction, a scavenger may be used, if necessary. Examples of the "scavenger" that can be used in this elimination reaction include ethanol, triisopropylsilane, 1-hydroxybenzotriazole, pyrrole, indole, 2,2,2-trifluoroethanol, methanol, anisole, mercaptoethanol, and thioanisole. The amount of the scavenger that can be used in this elimination reaction is suitably within the range of, for example, 1 to 100 times, and preferably 1 to 50 times, the molar ratio relative to 1 mole of the compound [C-1].

[0070] In compound [C-1], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (4) a substituent [7] (excluding the case where Z is a solid phase support), for example, the elimination reaction can be carried out by (1) stirring compound [C-1] and an acid in a suitable reaction solvent in a suitable reaction vessel, or (2) supplying a solution containing compound [C-1] and a solution containing an acid independently through supply channels into a flow reactor or reaction channel, and mixing the solutions in the flow reactor or the like. The solvent that can be used in this elimination reaction is not particularly limited, and for example, a mixed solvent of a polar solvent and a halogen-based solvent can be used. The ratio of the polar solvent in the mixed solvent of a polar solvent and a halogen-based solvent is 1.0% by weight, preferably 2.0% by weight, more preferably 3.0% by weight, and even more preferably 5.0% by weight. The upper limit is 90% by weight, preferably 75% by weight, more preferably 50% by weight, even more preferably 40% by weight, and particularly preferably 30% by weight. Furthermore, the upper and lower limits can be used in appropriate combination, and the ratio of the polar solvent in the mixed solvent of a polar solvent and a halogen-based solvent is, for example, appropriately within the range of 1% to 50%, preferably within the range of 1% to 40%, and particularly preferably within the range of 1% to 30%. Examples of the "acid" that can be used in this elimination reaction include the same as those mentioned above. When using these acids, they may be used in combination with a base (e.g., triethylamine) to adjust the acidity. The amount of the acid that can be used in this elimination reaction is suitably within the range of 1 to 500 times, and preferably 2 to 200 times, the molar ratio of which is based on 1 mole of the compound [C-1]. The acid that can be used in this elimination reaction is suitably diluted with a suitable solvent to a concentration within the range of, for example, 5% to 80%, and preferably diluted to a concentration within the range of 5% to 50%. The solvent for dissolving the acid that can be used in this elimination reaction is not particularly limited, but examples thereof include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, and mixed solvents thereof. In this step, a scavenger may be used, if necessary. Examples of the "scavenger" that can be used in this elimination reaction include the same as those described above. The amount of the scavenger that can be used in this elimination reaction is suitably within the range of, for example, 1 to 100 times, and preferably 1 to 50 times, the molar ratio relative to 1 mole of the compound [C-1]. Examples of means for supplying a liquid to a supply flow path that can be used in this elimination reaction include pumps that are commonly used in this field for supplying liquids, specifically, for example, a syringe pump, a plunger pump, a diaphragm pump, and a gear pump. Examples of flow reactors that can be used in the present elimination reaction include microreactors and in-line mixers such as static mixers. As a means for guiding the mixture from the supply channel to the reaction channel that can be used in this elimination reaction, for example, a multi-stage collision type micromixer can be mentioned. Examples of materials for the supply flow path and the reaction flow path that can be used in this elimination reaction include tubes made of synthetic resins selected from the group consisting of fluorine-based resins such as perfluoroalkoxyalkane (PFA), vinyl chloride-based resins, polyamide-based resins, and aromatic polyether ketone-based resins, and tubes made of metals selected from the group consisting of stainless steel, copper and its alloys, and titanium and its alloys. The inner diameter of the supply channel and reaction channel that can be used in this elimination reaction may be appropriately selected, for example, from a range of usually 0.1 mm to 1.0 mm, and is preferably selected, for example, from a range of 0.2 mm to 1.0 mm.

[0071] As described in the Test Examples and Examples below, in the method for producing compound [E-1], a reaction promoter is used, and a solution containing an acid is added to a reaction mixture containing compound [C-1] produced by subjecting compound [A-1] and compound [B-1] to a condensation reaction, thereby obtaining Q 1 The elimination reactions of the above can be carried out in the same system as the present continuous reaction. In addition, in the method for producing compound [C-1], a reaction promoter is used to remove Q of compound [A-1-1]. 1and then subjecting compound [A-1] and compound [B-1] to a condensation reaction in the same system to form compound [C-1]. The solvent that can be used in this 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. For example, aromatic solvents such as benzene, toluene, xylene, mesitylene, etc.; ester solvents such as ethyl acetate, isopropyl acetate, etc.; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, cyclohexane, etc. Two or more of these solvents may be mixed and used. Solvents that can be used in the present continuous reaction include, for example, polar solvents and halogenated solvents. The solvent that can be used in the continuous reaction is not particularly limited, but for example, a mixed solvent of a polar solvent and a halogen-based solvent can be used. The ratio of the polar solvent in the mixed solvent of the polar solvent and the halogen-based solvent is 1.0% by weight, preferably 2.0% by weight, more preferably 3.0% by weight, and even more preferably 5.0% by weight. The upper limit is 90% by weight, preferably 75% by weight, more preferably 50% by weight, even more preferably 40% by weight, and particularly preferably 30% by weight. Furthermore, the upper and lower limits can be used in appropriate combination, and the ratio of the polar solvent in the mixed solvent of the polar solvent and the halogen-based solvent is, for example, appropriately within the range of 1% to 50%, preferably within the range of 1% to 40%, and particularly preferably within the range of 1% to 30%.

[0072] This continuous reaction can be carried out, for example, In the presence of a reaction promoter, Formula [A-1-1]: [ka] [In the formula, B P is an optionally protected nucleobase; Q 1 is a group removable under acidic conditions; W is an oxygen atom or a sulfur atom; X is di(C1-6 alkyl)amino or a group represented by the general formulas [2-1] to [2-8]: [ka] [In the formula, * indicates the bonding position with P] The substituents are selected from those represented by , good Preferably it is di(C1-6 alkyl)amino, more preferably dimethylamino; G is a group represented by the general formula [7]: [ka] (In the formula, * indicates the binding position with T; Z is the general formula [8A]~[8D], [8E], [8G], [8H], [8J], [8K], [8N]: [ka] (In the formula, * indicates the bonding position with L; k represents an integer from 0 to 5; R 8a is a hydrogen atom or C 1-6 represents alkyl; R 8b are the same or different and each represents a long chain alkyl; R 8c are the same or different and have the following general formula [9A]: [ka] (In the formula, * represents the bond position; and R 9 represents a long chain alkyl and / or a long chain alkenyl. represents a substituent represented by the formula: R 8d are the same or different and represent a hydrogen atom, a halogen, a long-chain alkyl which may be substituted with 1 to 13 halogens, or a long-chain alkyloxy which may be substituted with 1 to 13 halogens; R 8e teeth, (1) Long-chain alkyl, (2) Long chain alkyl-carbonyl or (3) benzoyl substituted with 1 to 5 long chain alkyloxy and / or long chain alkenyloxy; and R 8f teeth, (1) Long-chain alkyl, (2) Long chain alkyl-carbonyl or (3) Long-chain alkenyl-carbonyl. is a substituent represented by the formula: L is the general formula

[10] : [ka] (In the formula, * represents the bond position to Z; ** indicates the bond position to the oxygen atom; and L 1 is optionally substituted C 2-10 Alkylene or optionally substituted C 6-10 It represents arylene. It is a substituent represented by the following formula: is a substituent represented by the formula: T is a single bond or a group of the following general formula

[11] : [ka] (In the formula, X and W are as defined above; * indicates the bond position to O; ** represents the bond position with G; and q represents an integer from 0 to 10. is a substituent represented by n is 1 to 25. From the compound, Q1 Detachment do, Formula [A-1]: [ka] [In the formula, BP , W, X, G, T, and n are as defined above.] The compound is Formula [B-1]: [ka] [In the formula, B P , Q 1 , W, X, G, and T are as defined above; D is a halogen; p is an integer from 1 to 10. By reacting with the compound Formula [C-1]: [ka] [where n, p, B P , Q 1 , W, X, G, and T are as defined above.] The present invention relates to a method for preparing a compound of formula (1).

[0073] This continuous reaction can be carried out, for example, In the presence of a reaction promoter, Formula [A-1-1]: [ka] [In the formula, Q 1 is trityl, monomethoxytrityl, or dimethoxytrityl, and n, B P , W, X, G, and T are as defined above.] From the compound in the presence of trifluoroacetic acid and 2,2,2-trifluoroethanol, optionally triisopropylsilane or ethanol; Q1 Detachment This may include: The continuous reaction can be carried out in a flow reactor. For example, A solution containing the compound of the general formula [A-1-1] and a solution containing an acid are supplied to a flow reactor to form Q1. Detachmentto obtain a compound of formula [A-1], and A method in which a solution containing the compound of the general formula [A-1] and a solution containing the compound of the general formula [B-1] are supplied to a subsequent flow reactor to prepare the compound of the general formula [C-1] is exemplified. In some cases, a flow reactor may be used which supplies a solution containing the compound of formula [A-1] and a solution containing a scavenger, or a flow reactor which supplies a solution containing an excess of the compound of formula [B-1] and the compound of formula [C-1], and a solution containing at least one selected from the group consisting of morpholine, 1-methylpiperazine, and N-ethylmorpholine.

[0074] (G-2) Q in the molecule of compound [C-2] 1 Method of detachment Since compound [C-2] is an unstable compound, the Q substituted at the 5'-position oxygen atom of the 5'-terminal nucleoside of compound [C-2] is 1 Before removing the phosphorus atom, it is preferable to first use an oxidizing agent to oxidize the phosphorus atom on the phosphorus bond formed in the condensation reaction from trivalent to pentavalent, thereby converting it into a compound represented by the following general formula [D-2] (hereinafter referred to as "compound [D-2]").

[0075] [ka] [In the formula, n, p, B P , G, Q 1 , R 4a , T, W and X are as defined above.]

[0076] Step 1: Preparation of compound [D-2] In the compound [C-2], when G is a substituent [7] and Z is a solid support, the oxidation reaction of the phosphorus atom can be carried out according to a method known per se (Current Protocols in Nucleic Acid Chemistry). Examples of oxidizing agents that can be used in this step include commercially available oxidizing solutions for nucleic acid synthesis [oxidizing solution-2, 0.1 mol / L iodine / 78% tetrahydrofuran / 20% pyridine / 2% water, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.; oxidizing solution, 0.5 M acetone solution of 0.5 M (1S)-(+)-(10-camphorsulfonyl)-oxaziridine, manufactured by Glen Research]. When a phosphorus atom is oxidized to form a phosphorothioate, the oxidation reaction of the phosphorus atom can be carried out according to a method known per se (see, for example, Current Protocols in Nucleic Acid Chemistry). Examples of the oxidizing agent that can be used in this step include commercially available sulfurizing reagents for nucleic acid synthesis [3-{(N,N-dimethylaminomethylidene)amino})-3H-1,2,4-dithiazole-5-thione (DDTT), manufactured by Glen Research; 5-phenyl-3H-1,2,4-dithiazole-3-one for nucleic acid synthesis, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.]. In this step, it is appropriate to use these oxidizing agents by dissolving them in a suitable solvent.

[0077] In compound [C-2], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (4) a substituent [7] (except when Z is a solid phase support), the oxidation reaction of the phosphorus atom can be carried out in accordance with a method known per se (see, for example, Nucleic Acids Research, Vol. 21, No. 5, pp. 1213-1217 (1993)). Examples of the oxidizing agent that can be used in this step include (+)-camphorylsulfonyloxaziridine (CSO), (+)-(8,8-dichlorocamphorylsulfonyl)-oxaziridine (DCSO), methyl ethyl ketone peroxide, and tert-butyl hydroperoxide (TBHP).

[0078] Step 2: Preparation of compound [E-2] By reacting compound [D-2] with an acid, the Q substituted at the 5'-position of the nucleoside unit on the 5'-terminal side of compound [D-2] is obtained. 1 can be eliminated from compound [D-2]. 1 By eliminating the above, a compound represented by the above general formula [E-2] (hereinafter referred to as "compound [E-2]") can be produced.

[0079] When G in compound [D-2] is a substituent [7] and Z is a solid phase support, the purification can be carried out, for example, by packing compound [D-2] into an appropriate column and eluting with a solution containing an acid, or by shaking or stirring a solution containing compound [D-2] and an acid in a reaction vessel equipped with a filter. Q in the molecule of compound [D-2] 1 The elimination reaction of can be carried out according to a method known per se (see, for example, Current Protocols in Nucleic Acid Chemistry). Acids that can be used in this step include, for example, commercially available deblocking solutions 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)].

[0080] In compound [D-2], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (4) a substituent [7] (excluding the case where Z is a solid phase support), for example, the reaction can be carried out by (1) stirring compound [D-2] and an acid in a suitable reaction solvent in a suitable reaction vessel, or (2) supplying a solution containing compound [D-2] and a solution containing an acid independently through a supply flow path into a flow reactor or a reaction flow path, and mixing the solutions in the flow reactor or the like. Q in the molecule of compound [D-2] 1The elimination reaction of can be carried out according to a method known per se (see, for example, Nucleic Acids Research, Vol. 21, No. 5, 1213-1217 (1993)). Acids that can be used in this step include, for example, dichloroacetic acid and trichloroacetic acid.

[0081] (H) Final deprotection and isolation of nucleic acid compounds When the compound [C-1], the compound [D-2], the compound [E-1] or the compound [E-2] has a protecting group in the molecule, a deprotection treatment according to the type or nature of the protecting group can be carried out to produce a compound from which all the protecting groups have been removed. For example, all the protecting groups of the compound can be removed according to the deprotection method described in "Green's PROTECTIVE GROUPS in ORGANIC SYNTHESIS, 4th Edition, 2006". Detachment Specifically, for example, by treating with (1) ammonia water, (2) ammonia water / ethanol, or (3) a mixed solution of ammonia water and methylamine aqueous solution, the protecting groups of the amino group or hydroxyl group of the nucleic acid base and the substituent [6] in the molecule of compound [C-1], compound [D-2], compound [E-1], or compound [E-2] can be removed. Detachment It is possible. In addition, for example, the Q 1 The same as the "acid" described in the "Method of Removal of Compound [C-2]" above, 1 The protecting group of the amino group at the 3'-position of the nucleoside at the 3'-end of compound [C-1] and the hydroxyl group at the 5'-position of the nucleoside at the 5'-end of compound [D-2] are substituted with a group that can be removed under acidic conditions by treating with the same "acid" described in "Step 2: Preparation of compound [E-2]" of "Method for removing nucleoside from compound [C-1]" or a solution of hydrochloric acid or acetic acid diluted with an appropriate solvent. Detachment It is possible. After removing the protecting group of the nucleic acid base portion, a group that can be removed under acidic conditions, which is substituted on the 5'-position hydroxyl group of the 5'-terminal nucleoside of compound [D-2], is added. DetachmentWhen the nucleic acid base moiety is protected, the acid is diluted with water and used. When the nucleic acid base moiety is protected, the acid is diluted with a suitable organic solvent.

[0082] (I) Purification and separation process All of the protecting groups in compound [C-1] or compound [E-1] are Detachment The separated compounds can be separated from the reaction mixture by conventional separation and purification methods, such as extraction, concentration, neutralization, filtration, centrifugation, recrystallization, C 8 From C 18 The protein can be isolated by using 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, either alone or in combination (see, for example, International Publication No. 1991 / 09033A1). When the desired compound is purified using reverse phase chromatography, for example, a mixed solution of 20 mM triethylamine / acetic acid buffer and acetonitrile can be used as the elution solvent. Furthermore, when ion exchange chromatography is used to purify the desired compound, for example, a mixture of 1 M saline and 10 mM aqueous sodium hydroxide solution or 0.3 M saline in 50 mM phosphate buffer can be used.

[0083] All of the protecting groups in compound [D-2] or compound [E-2] are Detachment The compound thus obtained can be separated and purified from the reaction mixture by conventional separation and purification methods, such as extraction, concentration, neutralization, filtration, centrifugation, recrystallization, C 8 From C 18 Reverse phase column chromatography, C 8 From C 18 The compound can be isolated and purified by using, alone or in combination, means such as reverse phase cartridge column, cation exchange column chromatography, anion exchange column chromatography, gel filtration column chromatography, high performance liquid chromatography, dialysis, and ultrafiltration. Examples of the "elution solvent" include acetonitrile, methanol, ethanol, isopropyl alcohol, or water alone, or a mixture of these solvents in any ratio. In this case, an additive such as sodium phosphate, potassium phosphate, sodium chloride, potassium chloride, ammonium acetate, triethylammonium acetate, sodium acetate, potassium acetate, tris hydrochloric acid, or ethylenediaminetetraacetic acid may be added at a concentration of 1 mM to 2 M to adjust the pH of the solution to within the range of 1 to 9.

[0084] (J) Preparation of compound [A] Compound [A] can be produced, for example, by introducing a substituent [6] into a hydroxyl group of a compound corresponding to compound [A] in accordance with a known method.

[0085] Representative examples are introduced below to explain the production method of compound [A].

[0086] (J-1) Production of compound [A-1] Compound [A] which is composed of one to more nucleoside units [4d] and in which the phosphorus bond between each nucleoside unit is a phosphorus bond [5], can be produced, for example, according to the methods described in (i) to (iv) below.

[0087] (i) Preparation of compound [A-1], wherein G is a silicon substituent and T is a single bond [ka] [In the formula, n, B P , Q 1 , X and W are as defined above; Hal stands for halogen; G 1 represents a silicon substituent. The compound represented by the above general formula [A-1a] (hereinafter referred to as "compound [A-1a]") is compound [A-1] in which G is a silicon substituent and T is a single bond. An example of a method for producing the compound [A-1a] is described below.

[0088] Step 1: Preparation of the compound represented by the above general formula [A-1a-Q1] (hereinafter referred to as "compound [A-1a-Q1]") Compound [A-1a-Q1] can be produced by introducing a silicon substituent into the hydroxyl group at the 5' end of compound

[21] using a compound [20A] (hereinafter referred to as "compound [20A]") represented by the above general formula

[21] (hereinafter referred to as "compound

[21] "). This reaction for introducing a silicon substituent can be carried out according to a method known per se.

[0089] Step 2: Preparation of compound [A-1a] Compound [A-1a] can be produced by treating compound [A-1a-Q1] with an acid. The "acid" that can be used in this step is the above-mentioned "Q 1 Examples of the "acid" include the same as those described in "Method for removing the acid". The amount of the acid that can be used in this step is suitably within the range of 1 to 500 times, and preferably 2 to 200 times, the molar ratio of which is based on 1 mole of the compound [A-1a-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, which is not particularly limited, and examples thereof include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixture thereof.

[0090] In this step, a scavenger may be used, if necessary. The "scavenger" that can be used in this step is the Q 1 Examples of the "scavenger" include the same ones as those described in "Method for removing the scavenger". The amount of the scavenger that can be used in this step is suitably within the range of 1 to 100 times, and preferably 1 to 50 times, the molar ratio of 1 mole of the compound [A-1a-Q1].

[0091] (ii) Preparation of compound [A-1], in which G is (1) a long-chain alkyl-carbonyl, (2) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (3) a substituent [7], and T is a single bond. [ka] [In the formula, n, B P , Q 1 X and W are as defined above; G 2 represents (1) a long chain alkyl-carbonyl, (2) a benzoyl substituted with 1 to 5 long chain alkyloxy and / or long chain alkenyloxy, or (3) a substituent [7]; and Y represents a hydroxyl group or a halogen.

[0092] The compound represented by the above general formula [A-1b] (hereinafter referred to as "compound [A-1b]") is a compound [A-1] in which G is (1) a long-chain alkyl-carbonyl, (2) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (3) a substituent [7], and T is a single bond. An example of a method for producing the compound [A-1b] is described below.

[0093] Step 1: Preparation of the compound represented by the above general formula [A-1b-Q1] (hereinafter referred to as "compound [A-1b-Q1]") Compound

[21] can be condensed with a compound represented by the above general formula [20B] (hereinafter referred to as "compound [20B]") to produce compound [A-1b-Q1]. The condensation reaction can be carried out according to a method known per se. When a compound [20B] in which Y is a hydroxyl group is used in this step, the reaction can be carried out in the presence or absence of a base using a condensing agent at a temperature within the range of -20°C to 100°C. When a compound [20B] in which Y is a halogen is used in this step, the reaction can be carried out in the presence of a base at a temperature within the range of -20°C to 100°C. Examples of condensing agents that can be used in this step include 1,1'-oxalyldiimidazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, dicyclohexylcarbodiimide, diethyl cyanophosphonate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate. Examples of the base that can be used in this step include organic bases such as triethylamine, N,N-diisopropylethylamine, N,N-dimethylaniline, pyridine, and 1,8-diazabicyclo[5,4,0]-7-undecene. The solvent that can be used in this step is not particularly limited, but examples thereof include ethers such as THF, 1,4-dioxane, diethyl ether, etc.; amides such as dimethylformamide, dimethylacetamide, etc.; nitriles such as acetonitrile, propionitrile, etc.; hydrocarbons such as benzene, toluene, etc.; halogenated hydrocarbons such as chloroform, methylene chloride, etc.; and mixed solvents thereof. When the compound [20B] in which Y is a hydroxyl group is used in this step, an additive can be used, if necessary. Additives that can be used in this step include, for example, 4-dimethylaminopyridine, 1-hydroxybenzotriazole, and 1-hydroxy-7-azabenzotriazole. The reaction time varies depending on the types of raw materials used, the reaction temperature, etc., but is usually within the range of 10 minutes to 24 hours. The amounts of compound

[21] and the condensing agent used are suitably within a range of, for example, 1 to 1.5 moles per mole of compound [20B]. The amount of the base used is, for example, within the range of 1 to 10 equivalents, and preferably within the range of 1 to 4 equivalents, relative to compound [20B].

[0094] Step 2: Preparation of compound [A-1b] Compound [A-1b] can be produced by treating compound [A-1b-Q1] with an acid.

[0095] The "acid" that can be used in this step is the above-mentioned "Q 1 Examples of the "acid" include the same as those described in "Method for removing the acid". The amount of the acid that can be used in this step is suitably within the range of 1 to 500 times, and preferably 2 to 200 times, the molar ratio of which is based on 1 mole of the compound [A-1b-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, which is not particularly limited, and examples thereof include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixture thereof.

[0096] In this step, a scavenger may be used, if necessary. The "scavenger" that can be used in this step is the Q 1 Examples of the "scavenger" include the same ones as those described in "Method for removing the scavenger". The amount of the scavenger that can be used in this step is suitably within the range of 1 to 100 times, and preferably 1 to 50 times, the molar ratio of 1 mole of the compound [A-1b-Q1].

[0097] (iii) Preparation of compound [A-1], in which G is (1) a long-chain alkyl-carbonyl, (2) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (3) a substituent [7], and T is a substituent

[11] . [ka] [In the formula, n, q, B P , D, G 2 , Q 1 , T, X and W are as defined above; Trt is trityl.

[0098] The compound represented by the above general formula [A-1c] (hereinafter referred to as "compound [A-1c]") is a compound [A-1] in which G is (1) a long-chain alkyl-carbonyl, (2) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (3) a substituent [7], and T is a substituent

[11] . An example of a method for producing compound [A-1c] is described below.

[0099] Step 1: Preparation of the compound represented by the above general formula

[23] (hereinafter referred to as "compound

[23] ") Compound

[23] can be produced by condensing a compound represented by the above general formula [20C] (hereinafter referred to as "compound [20C]") with a compound represented by the above general formula

[22] (hereinafter referred to as "compound

[22] "). Compound [20C] is a carboxylic acid, but its reactive derivative can also be used in this step. Examples of the reactive derivative of compound [20C] include those that are commonly used in ester condensation reactions, such as acid halides (e.g., acid chlorides, acid bromides). Compound

[22] can be produced in accordance with a known method (see, for example, U.S. Patent Application Publication No. 2014 / 0330006A1). In addition, according to a known method (for example, see International Publication No. 2014 / 077292A1), compound [20C] in which G is benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy can be produced.

[0100] Step 2: Preparation of the compound represented by the above general formula

[24] (hereinafter referred to as "compound

[24] ") Compound

[24] can be produced by removing the trityl group in the compound

[23] with an acid.

[0101] Step 3: Preparation of the compound represented by the above general formula [A-1c-Q1] (hereinafter referred to as "compound [A-1c-Q1]") Compound

[24] and the compound represented by the above general formula

[25] (hereinafter referred to as "compound

[25] ") can be condensed to produce compound [A-1c-Q1]. The condensation reaction and deprotection reaction can be carried out according to a method known per se.

[0102] Step 4: Preparation of compound [A-1c] Compound [A-1c] can be produced by treating compound [A-1c-Q1] with an acid. The "acid" that can be used in this step is the above-mentioned "Q 1 Examples of the "acid" include the same as those described in "Method for removing the acid". The amount of the acid that can be used in this step is suitably within the range of 1 to 500 times, and preferably 2 to 200 times, the molar ratio of which is based on 1 mole of the compound [A-1c-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, which is not particularly limited, and examples thereof include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixture thereof.

[0103] In this step, a scavenger may be used, if necessary. The "scavenger" that can be used in this step is the Q 1 Examples of the "scavenger" include the same ones as those described in "Method for removing the scavenger". The amount of the scavenger that can be used in this step is suitably within the range of, for example, 1 to 100 times, and preferably 1 to 50 times, the molar ratio of 1 mole of the compound [A-1c-Q1].

[0104] (iv) Preparation of compound [A-1], wherein G is a substituent [7] and T is a single bond [ka] [In the formula, n, B P , L 1 , Q 1 , X, W and Z are as defined above.]

[0105] The compound represented by the above general formula [A-1d] (hereinafter referred to as "compound [A-1d]") is compound [A-1] in which G is a substituent [7] and T is a single bond. An example of a method for producing compound [A-1d] is described below.

[0106] Step 1: Preparation of the compound represented by the above general formula [A-1d-Q1] (hereinafter referred to as "compound [A-1d-Q1]") The compound represented by the above general formula [20D] (hereinafter referred to as "compound [20D]") can be condensed with the compound represented by the above general formula

[21] (hereinafter referred to as compound

[21] ) to produce compound [A-1d-Q1]. The condensation reaction can be carried out in accordance with a method known per se. Compound [20D] is a carboxylic acid, but its reactive derivative can also be used in this step. Examples of the reactive derivative of compound [20D] include those that are commonly used in ester condensation reactions, such as acid halides (e.g., acid chlorides, acid bromides). When compound [20D] is used, the reaction can be carried out in the presence or absence of a base using a condensing agent at a temperature within the range of -20°C to 100°C. Examples of condensing agents that can be used in this step include 1,1'-oxalyldiimidazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, dicyclohexylcarbodiimide, diethyl cyanophosphonate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate. Examples of the base that can be used in this step include organic bases such as triethylamine, N,N-diisopropylethylamine, N,N-dimethylaniline, pyridine, and 1,8-diazabicyclo[5,4,0]-7-undecene. Solvents that can be used in this step include, but are not limited to, ethers such as THF, 1,4-dioxane, diethyl ether, etc., amides such as dimethylformamide, dimethylacetamide, etc., nitriles such as acetonitrile, propionitrile, etc., hydrocarbons such as benzene, toluene, etc., halogenated hydrocarbons such as chloroform, methylene chloride, etc., or mixed solvents thereof. In addition, additives can be used as necessary. Additives that can be used in this step include, for example, 4-dimethylaminopyridine, 1-hydroxybenzotriazole, and 1-hydroxy-7-azabenzotriazole. The reaction time varies depending on the types of raw materials used, the reaction temperature, etc., but is usually within the range of 10 minutes to 24 hours. The amounts of compound

[21] and the condensing agent used are suitably within the range of, for example, 1 to 1.5 moles per mole of compound [20D]. The amount of the base used is, for example, within the range of 1 to 10 equivalents, preferably 1 to 4 equivalents, relative to compound [20D].

[0107] Step 2: Preparation of compound [A-1d] Compound [A-1d] can be produced by treating compound [A-1d-Q1] with an acid. The "acid" that can be used in this step is the above-mentioned "Q 1 Examples of the "acid" include the same as those described in "Method for removing the acid". The amount of the acid that can be used in this step is suitably within the range of 1 to 500 times, and preferably 2 to 200 times, the molar ratio of which is based on 1 mole of the compound [A-1d-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, which is not particularly limited, and examples thereof include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixture thereof. In this step, a scavenger may be used, if necessary. The "scavenger" that can be used in this step is the Q 1 Examples of the "scavenger" include the same ones as those described in "Method for removing the scavenger". The amount of the scavenger that can be used in this step is suitably within the range of 1 to 100 times, and preferably 1 to 50 times, the molar ratio of 1 mole of the compound [A-1d-Q1].

[0108] For example, compound [20D] can be produced according to the process described below. [ka] [In the formula, L 1 and Z is as defined above; R is C 1-6 represents alkyl.

[0109] Step 1: Preparation of the compound represented by the above general formula

[28] (hereinafter referred to as "compound

[28] ") Compound

[28] can be produced by condensing the compound represented by the above general formula

[26] (hereinafter referred to as "compound

[26] ") with the compound represented by the above general formula

[27] (hereinafter referred to as "compound

[27] "). The condensation reaction can be carried out in accordance with a method known per se. The reagents and reaction conditions that can be used in this step can be the same as those in the above "Production of compound [A-1b-Q1]".

[0110] Step 2: Preparation of compound [20D] Compound [20D] can be produced by ester hydrolysis of compound

[28] . The ester hydrolysis reaction can be carried out in accordance with a method known per se. The solvent that can be used in this step is not particularly limited, but examples thereof include water, alcohols such as methanol and ethanol, ethers such as tetrahydrofuran, 1,4-dioxane, and diethyl ether, nitriles such as acetonitrile and propionitrile, hydrocarbons such as benzene and toluene, halogenated hydrocarbons such as chloroform and methylene chloride, and mixed solvents thereof. This step is carried out in the presence of a base such as sodium hydroxide, potassium hydroxide, or lithium hydroxide at a temperature within the range of 20°C to 100°C. The reaction time varies depending on the types of raw materials used, the reaction temperature, etc., but is usually within the range of 10 minutes to 24 hours.

[0111] Moreover, the compound

[26] can be produced, for example, according to the methods described in the following (a) to (j). (a) For example, by using commercially available primary amine compounds or by amminating commercially available alkyl halides, Z is a substituent [8A] and R 8a is a hydrogen atom, and R 8b Compound

[26] can be prepared in which is a long chain alkyl. (b) For example, by alkylating a commercially available primary amine compound, Z is a substituent [8A] or a substituent [8B], and R 8a C 1-6 is alkyl, R 8b are the same or different and are long-chain alkyl groups, the compound

[26] can be produced. The alkylation reaction can be carried out according to a method known per se. (c) Compound

[26] in which Z is a substituent [8C] can be produced according to known methods (see, for example, Cancer Res., 2008 Nov 1; 68(21):8843-8851, Chem. Sci., 2016, 7, 2308-2321). (d) For example, compound

[26] in which Z is a substituent [8D] can be produced by condensing methyl phthalate with 1-(tert-butoxycarbonyl)piperazine, hydrolyzing the ester moiety using an alkali such as sodium hydroxide, and then condensing with compound

[26] in which Z is a substituent [8A], followed by removing the tert-butoxycarbonyl group using an acid such as trifluoroacetic acid. The condensation reaction, the hydrolysis reaction using an alkali, and the deprotection reaction of the tert-butoxycarbonyl group using an acid can be carried out in accordance with a method known per se. (e) For example, one hydroxyl group of ethane-1,2-diol can be alkylated with an alkyl halide to give a compound in which Z is a substituent [8E] and R 8e is a long chain alkyl group, compound

[26] can be prepared. For example, one hydroxyl group of ethane-1,2-diol can be converted to a long-chain alkyl-carbonyl group, where Z is a substituent [8E] and R 8e Compound

[26] can be produced in which is a long-chain alkyl-carbonyl. The compound used for the long-chain alkyl-carbonylation can be, for example, the corresponding carboxylic acid compound or a reactive derivative thereof. The reactive derivative can be, for example, an acid halide (e.g., acid chloride, acid bromide), or the like, which is usually used in an ester condensation reaction. For example, one hydroxyl group of ethane-1,2-diol can be condensed with compound [20C] to give a compound in which Z is a substituent [8E] and R 8e is a benzoyl group substituted with 1 to 5 long chain alkyloxy and / or long chain alkenyloxy groups, it is possible to prepare a compound

[26] . (f) For example, compound

[26] in which Z is a substituent [8F] can be prepared according to a method similar to that for preparing compound (18) in which Z is a substituent [8E], using 2-amino-ethanol instead of ethane-1,2-diol. (g) For example, compound

[26] in which Z is a substituent [8G] can be produced by condensing 9-fluorenylmethyloxycarbonyl-phenylalanine with compound

[26] in which Z is a substituent [8A], and then removing the 9-fluorenylmethyloxycarbonyl group with piperidine. The condensation reaction and the deprotection reaction of the 9-fluorenylmethyloxycarbonyl group can be carried out in accordance with a method known per se. (h) For example, compound

[26] in which Z is a substituent [8H] can be produced by the same method as in the production method of compound

[26] in which Z is a substituent [8E] using 1-(tert-butoxycarbonyl)piperazine instead of ethane-1,2-diol, and then deprotecting the tert-butoxycarbonyl group in the molecule with an acid. (i) According to a known method (see, for example, Japanese Patent No. 5705512, Tetrahedron Letters, Vol. 53, 1936-1939 (2012), International Publication No. 2014 / 189142A1, International Publication No. 2016 / 060135A1, International Publication No. 2016 / 140232A1), Z is the substituent [8I], the substituent [8J], the substituent [8K], the substituent [8L], the substituent [8N], the compound

[26] can be produced. (j) For example, 9H-xanthene-9-one having a hydroxyl group can be treated with a base such as sodium hydride, and then treated with an appropriate long-chain alkyl halide to produce the corresponding 9H-xanthene-9-one having a long-chain alkyloxy. Compound

[26] in which Z is a substituent group [8M] can be produced by further treating with optionally substituted phenylmagnesium bromide. In addition, compound

[26] in which Z is a substituent group [8M] can be produced by adjusting 9H-xanthene-9-one derivatives or phenylmagnesium bromide derivatives having various substituent groups according to known methods.

[0112] In addition, compound

[21] where n=1 can be prepared according to known methods (see, for example, International Publication No. 91 / 09033A1), and compound

[21] where n>1 can be prepared according to the method described below. [ka] [In the formula, n, B P , D, Q 1 , X and W are as defined above; Ac stands for acetyl.

[0113] Step 1: Preparation of the compound represented by the above general formula

[30] (hereinafter referred to as "compound

[30] ") The compound represented by the above general formula

[29] (see, for example, International Publication WO 91 / 09033A1) is acetylated with acetic anhydride in the presence of a base to produce the compound

[30] . The acetylation reaction can be carried out according to a method known per se.

[0114] Step 2: Preparation of the compound represented by the above general formula

[31] (hereinafter referred to as "compound

[31] ") Compound

[31] can be produced by treating compound

[30] with an acid. 1 The elimination reaction of can be carried out in accordance with a method known per se.

[0115] Step 3: Preparation of the compound represented by the above general formula

[33] (hereinafter referred to as "compound

[33] ") Compound

[33] can be produced by condensing compound

[31] with a compound represented by the above general formula

[32] (hereinafter referred to as "compound

[32] "). The condensation reaction can be carried out in accordance with a method known per se (see, for example, International Publication No. 91 / 09033A1). Compound

[32] can be produced, for example, according to a known method (see, for example, International Publication No. 91 / 09033A1).

[0116] Step 4: Preparation of compound

[21] For example, the acetyl group of compound

[33] can be selectively eliminated using an alkali metal alkoxide such as sodium methoxide to produce compound

[21] . The acetyl elimination reaction can be carried out in accordance with a method known per se (see, for example, Tetrahedron Letters, Vol. 50, 1751-1753 (2009)).

[0117] (J-2) Production of compound [A-2] Compound [A], which is composed of one to more nucleoside units selected from the group consisting of nucleoside unit [4a], nucleoside unit [4b] and nucleoside unit [4c], and in which the phosphorus bond between each nucleoside unit is phosphorus bond [5], can be produced, for example, by the methods described in (i) to (iv) below.

[0118] (i) Preparation of compound [A-2], wherein G is a silicon substituent and T is a single bond [ka] [In the formula, n, B P , G 1 , Hal, Q. 1 , R 4a , X and W are as defined above.] The compound represented by the above general formula [A-2a] (hereinafter referred to as "compound [A-2a]") is compound [A-2] in which G is a silicon substituent and T is a single bond. An example of a method for producing the compound [A-2a] is described below.

[0119] Step 1: Preparation of the compound represented by the above general formula [A-2a-Q1] (hereinafter referred to as "compound [A-2a-Q1]") Compound [A-2a-Q1] can be produced by introducing a silicon substituent into the 3'-position hydroxyl group of the nucleoside unit at the 3'-end of compound

[34] (hereinafter referred to as "compound

[34] ") using compound [20A]. This reaction for introducing a silicon substituent can be carried out according to a method known per se.

[0120] Step 2: Preparation of compound [A-2a] Compound [A-2a] can be produced by treating compound [A-2a-Q1] with an acid. The "acid" that can be used in this step is the Q 1 Examples of the "acid" include the same as those described in the "Elimination reaction of the aryl group". The amount of the acid that can be used in this step is suitably within the range of 1 to 500 times, and preferably 2 to 200 times, the molar ratio of which is based on 1 mole of the compound [A-2a-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, which is not particularly limited, and examples thereof include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixture thereof.

[0121] In this step, a scavenger may be used, if necessary. The "scavenger" that can be used in this step is the Q 1 Examples of the "scavenger" include the same ones as those described in the "elimination reaction of the scavenger". The amount of the scavenger that can be used in this step is suitably within the range of, for example, 1 to 100 times, and preferably 1 to 50 times, the molar ratio relative to 1 mole of the compound [A-2a-Q1].

[0122] (ii) Preparation of compound [A-2], in which G is (1) a long-chain alkyl-carbonyl, (2) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (3) a substituent [7], and T is a single bond. [ka] [In the formula, n, B P , G 2 , Q 1 , R 4a , X, Y and W are as defined above.]

[0123] The compound represented by the above general formula [A-2b] (hereinafter referred to as "compound [A-2b]") is compound [A-2] in which G is (1) long-chain alkyl-carbonyl, (2) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (3) substituent [7], and T is a single bond.

[0124] An example of a method for producing the compound [A-2b] is described below.

[0125] Step 1: Preparation of the compound represented by the above general formula [A-2b-Q1] (hereinafter referred to as "compound [A-2b-Q1]") Compound [20B] can be condensed with compound

[34] to produce compound [A-2b-Q1]. The condensation reaction can be carried out according to a method known per se. When a compound [20B] in which Y is a hydroxyl group is used in this step, the reaction can be carried out in the presence or absence of a base using a condensing agent at a temperature within the range of -20°C to 100°C. When a compound [20B] in which Y is a halogen is used in this step, the reaction can be carried out in the presence of a base at a temperature within the range of -20°C to 100°C. Examples of condensing agents that can be used in this step include 1,1'-oxalyldiimidazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, dicyclohexylcarbodiimide, diethyl cyanophosphonate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate. Examples of the base that can be used in this step include organic bases such as triethylamine, N,N-diisopropylethylamine, N,N-dimethylaniline, pyridine, and 1,8-diazabicyclo[5,4,0]-7-undecene. The solvent that can be used in this step is not particularly limited, but examples thereof include ethers such as THF, 1,4-dioxane, diethyl ether, etc.; amides such as dimethylformamide, dimethylacetamide, etc.; nitriles such as acetonitrile, propionitrile, etc.; hydrocarbons such as benzene, toluene, etc.; halogenated hydrocarbons such as chloroform, methylene chloride, etc.; and mixed solvents thereof. When the compound [20B] in which Y is a hydroxyl group is used in this step, an additive can be used, if necessary. Examples of additives that can be used include 4-dimethylaminopyridine, 1-hydroxybenzotriazole, and 1-hydroxy-7-azabenzotriazole. The reaction time varies depending on the types of raw materials used, the reaction temperature, etc., but is usually within the range of 10 minutes to 24 hours. The amounts of compound [20B] and the condensing agent used are suitably within a range of, for example, 1 to 1.5 moles per mole of compound

[34] . The amount of the base used is, for example, within the range of 1 to 10 equivalents, preferably 1 to 4 equivalents, relative to the compound

[34] .

[0126] Step 2: Preparation of compound [A-2b] Compound [A-2b] can be produced by treating compound [A-2b-Q1] with an acid.

[0127] The "acid" that can be used in this step is the Q 1 Examples of the "acid" include the same as those described in the "Elimination reaction of the aryl group". The amount of the acid that can be used in this step is suitably within the range of 1 to 500 times, and preferably 2 to 200 times, the molar ratio of which is based on 1 mole of the compound [A-2b-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, which is not particularly limited, and examples thereof include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixture thereof.

[0128] In this step, a scavenger may be used, if necessary. The "scavenger" that can be used in this step is the Q 1 Examples of the "scavenger" include the same ones as those described in the "elimination reaction of the scavenger". The amount of the scavenger that can be used in this step is suitably within the range of, for example, 1 to 100 times, and preferably 1 to 50 times, the molar ratio relative to 1 mole of the compound [A-2b-Q1].

[0129] (iii) Preparation of compound [A-2], in which G is (1) a long-chain alkyl-carbonyl, (2) a benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (3) a substituent [7], and T is a substituent

[11] . [ka] [In the formula, n, q, B P , D, G 2 , Q 1 , R 4a , T, X and W are as defined above.]

[0130] The compound represented by the above general formula [A-2c] (hereinafter referred to as "compound [A-2c]") is compound [A-2] in which G is (1) long-chain alkyl-carbonyl, (2) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (3) substituent [7], and T is substituent

[11] .

[0131] An example of a method for producing the compound [A-2c] is described below.

[0132] Step 1: Preparation of the compound represented by the above general formula

[36] (hereinafter referred to as "compound

[36] ") Compound

[36] can be produced by condensing compound

[24] with the compound represented by the above general formula

[35] (hereinafter referred to as "compound

[35] "). The condensation reaction and deprotection reaction can be carried out according to a method known per se.

[0133] Step 2: Preparation of the compound represented by the above general formula [A-2c-Q1] (hereinafter referred to as "compound [A-2c-Q1]") Compound [A-2c-Q1] can be produced by reacting compound

[36] with an oxidizing agent. The oxidation reaction can be carried out in accordance with a method known per se. Examples of the "oxidizing agent" include iodine and tert-butyl hydroperoxide. The oxidizing agent that can be used in this step can be diluted with a suitable solvent to a concentration of 0.05 to 2M. The solvent is not particularly limited, but examples include pyridine, tetrahydrofuran, water, and a mixture of these. For example, iodine / water / pyridine-tetrahydrofuran or iodine / pyridine-acetic acid, or a peroxidizing agent (such as tert-butyl hydroperoxide / methylene chloride) can be used. The reaction temperature is preferably 20°C to 50°C. The reaction time varies depending on the type of oxidizing agent used and the reaction temperature, but is usually 1 to 30 minutes. The amount of the oxidizing agent used is preferably 1 to 100 times, more preferably 10 to 50 times, the molar amount of the compound

[36] .

[0134] Step 3: Preparation of compound [A-2c] Compound [A-2c] can be produced by treating compound [A-2c-Q1] with an acid. The "acid" that can be used in this step is the Q 1 Examples of the "acid" include the same as those described in the "Elimination reaction of the aryl group". The amount of the acid that can be used in this step is, for example, within the range of 1 to 500 times, and preferably 2 to 200 times, the amount of the compound [A-2c-Q1] per 1 mole. The acid that can be used in this step may be diluted with an appropriate solvent, which is not particularly limited, and examples thereof include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixture thereof. In this step, a scavenger may be used, if necessary. The "scavenger" that can be used in this step is the Q 1 Examples of the "scavenger" include the same ones as those described in the "elimination reaction of the scavenger". The amount of the scavenger that can be used in this step is suitably within the range of 1 to 100 times, and preferably 1 to 50 times, the molar ratio of 1 mole of the compound [A-2c-Q1].

[0135] (iv) Preparation of compound [A-2], in which G is a substituent [7] and T is a single bond [ka] [In the formula, n, B P , L 1 , Q 1 , R 4a , X, W and Z are as defined above.] The compound represented by the above general formula [A-2d] (hereinafter referred to as "compound [A-2d]") is compound [A-2] in which G is a substituent [7] and T is a single bond.

[0136] An example of a method for producing the compound [A-2d] is described below.

[0137] Process 1 The compound represented by the above general formula [A-2d-Q1] (hereinafter referred to as "compound [A-2d-Q1]") can be produced by condensing compound [20D] with compound

[34] . The condensation reaction can be carried out according to a method known per se. Compound [20D] is a carboxylic acid, but its reactive derivative can also be used in this step. Examples of the reactive derivative of compound [20D] include those that are commonly used in ester condensation reactions, such as acid halides (e.g., acid chlorides, acid bromides). When compound [20D] is used, the reaction can be carried out in the presence or absence of a base using a condensing agent at a temperature within the range of -20°C to 100°C. Examples of condensing agents that can be used in this step include 1,1'-oxalyldiimidazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, dicyclohexylcarbodiimide, diethyl cyanophosphonate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate. Examples of the base that can be used in this step include organic bases such as triethylamine, N,N-diisopropylethylamine, N,N-dimethylaniline, pyridine, and 1,8-diazabicyclo[5,4,0]-7-undecene. Solvents that can be used in this step include, but are not limited to, ethers such as THF, 1,4-dioxane, diethyl ether, etc., amides such as dimethylformamide, dimethylacetamide, etc., nitriles such as acetonitrile, propionitrile, etc., hydrocarbons such as benzene, toluene, etc., halogenated hydrocarbons such as chloroform, methylene chloride, etc., or mixed solvents thereof. In addition, additives can be used as necessary. Additives that can be used in this step include, for example, 4-dimethylaminopyridine, 1-hydroxybenzotriazole, and 1-hydroxy-7-azabenzotriazole. The reaction time varies depending on the types of raw materials used, the reaction temperature, etc., but is usually within the range of 10 minutes to 24 hours. The amounts of compound [20D] and the condensing agent used are suitably within a range of, for example, 1 to 1.5 moles per mole of compound

[34] . The amount of the base used is, for example, within the range of 1 to 10 equivalents, preferably 1 to 4 equivalents, relative to the compound

[34] .

[0138] Process 2 Compound [A-2d] can be produced by treating compound [A-2d-Q1] with an acid. The "acid" that can be used in this step is the Q 1 Examples of the "acid" include the same as those described in the "Elimination reaction of the aryl group". The amount of the acid that can be used in this step is, for example, within the range of 1 to 500 times, and preferably 2 to 200 times, the amount of the compound [A-2d-Q1] per 1 mole. The acid that can be used in this step may be diluted with an appropriate solvent, which is not particularly limited, and examples thereof include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixture thereof. In this step, a scavenger may be used, if necessary. The "scavenger" that can be used in this step is the Q 1 Examples of the "scavenger" include the same ones as those described in the "elimination reaction of the scavenger". The amount of the scavenger that can be used in this step is suitably within the range of 1 to 100 times, and preferably 1 to 50 times, the molar ratio of 1 mole of the compound [A-2d-Q1].

[0139] In addition, compound

[34] where n=1 can be prepared according to known methods (Current Protocols in Nucleic Acid Chemistry), and compound

[34] where n>1 can be prepared according to known methods (see, for example, U.S. Patent Application Publication No. 2010 / 273999A1).

[0140] Compound "A-2" is a compound in which each nucleoside unit constituting said compound is nucleoside unit [4a]. Compounds in which all or part of nucleoside unit [4a] is replaced with nucleoside unit [4b] or nucleoside unit [4c] can also be produced by using a method similar to that described above.

[0141] (K) Production of compound [B] Compound [B] can be produced, for example, by introducing a substituent [1] into a hydroxyl group of a compound corresponding to compound [B] in accordance with a known method.

[0142] Representative examples are introduced below to explain the production method of compound [B].

[0143] (K-1) Production of compound [B-1] Compound [B] which is composed of one to more nucleoside units [4h] and in which the phosphorus bond between each nucleoside unit is a phosphorus bond [5], can be produced, for example, according to the method described below. [ka] [In the formula, p, B P , Q 1 , D, X and W are as defined above.]

[0144] Step 1: Preparation of the compound represented by the above general formula

[39] (hereinafter referred to as "compound

[39] ") Compound

[39] can be produced by condensing a compound represented by the above general formula

[37] (hereinafter referred to as "compound

[37] ") with a compound represented by the above general formula

[38] (hereinafter referred to as "compound

[38] "). The condensation reaction can be carried out in accordance with a method known per se (see, for example, U.S. Patent Application Publication No. 2014 / 0330006A1, WO 2012 / 043730A1, and WO 2013 / 082548A1).

[0145] Step 2: Preparation of compound [B-1] Compound [B-1] can be produced by condensing the compound represented by the above general formula

[40] (hereinafter referred to as "compound

[40] ") with compound

[39] . The condensation reaction can be carried out in accordance with a method known per se (see, for example, U.S. Patent Application Publication No. 2014 / 0330006A1, WO 2012 / 043730A1, WO 2013 / 082548A1, and WO 91 / 09033A1). Compound

[40] can be produced by the same method as that for producing compound

[21] .

[0146] (K-2) Production of compound [B-2] Compound [B], which is composed of one to more nucleoside units selected from the group consisting of nucleoside unit [4e], nucleoside unit [4f] and nucleoside unit [4g], and in which the phosphorus bond between each nucleoside unit is phosphorus bond [5], can be produced, for example, by the method described below. [ka] [In the formula, p, B P , Hal, Q. 1 ,D.R. 4a , X and W are as defined above.]

[0147] Step 1: Preparation of the compound represented by the above general formula

[42] (hereinafter referred to as "compound

[42] ") Compound

[42] can be produced by reacting compound

[37] with a compound represented by the above general formula

[41] (hereinafter referred to as "compound

[41] "). The reaction can be carried out in accordance with a method known per se (see, for example, Helvetica Chimica Acta, Vol. 70, 175-186 (1987), International Publication No. 2003 / 106468A1, Acta Nature, 6, 116-118 (2014), Russian Journal of General Chemistry, Vol. 67, No. 1, 62-64 (1997)).

[0148] Step 2: Preparation of compound [B-2] According to a known method, compound "B-2" can be produced by reacting compound

[42] with a compound represented by the above general formula

[43] (hereinafter referred to as "compound

[43] ") to introduce a substituent containing a phosphorus atom into the 3'-position hydroxyl group of the 3'-terminal nucleoside unit. In this step, an activator can be used, if necessary. The solvent used in this step is not particularly limited, but examples thereof include acetonitrile and tetrahydrofuran. The amount of the compound

[42] used is suitably 1 to 20 times, and preferably 1 to 10 times, the molar amount of the compound

[43] . Examples of the "activator" include 1H-tetrazole, 5-ethylthiotetrazole, 4,5-dichloroimidazole, 4,5-dicyanoimidazole, benzotriazole triflate, imidazole triflate, pyridinium triflate, N,N-diisopropylethylamine, and 2,4,6-collidine / N-methylimidazole. The amount of the "activator" used is suitably 1 to 20 times the molar amount, preferably 1 to 10 times the molar amount, relative to compound

[43] . The reaction temperature is suitably 0°C to 120°C. The reaction time varies depending on the types of raw materials used, the reaction temperature, etc., but is usually suitably 30 minutes to 24 hours.

[0149] Compound [B-2] is a compound in which each nucleoside unit constituting the compound is nucleoside unit [4e], but a compound in which all or part of nucleoside unit [4e] is replaced by nucleoside unit [4f] or nucleoside unit [4g] can also be produced according to the same method as above.

Examples

[0150] The present invention will be described in more detail below with reference to Examples, Comparative Examples, and Test Examples, but the present invention is not limited thereto. Note that the "conversion yield (%)" means the ratio of the raw material converted into the target product, and is calculated as "{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 is dissolved in acetonitrile or an 80% aqueous acetonitrile solution, and HPLC analysis is performed under the following conditions. The coupling efficiency was calculated by using the integrated value of the peak area obtained by absorption at UV = 264 nm in HPLC. <ODS conditions> Column: Waters XBridge C8 (5μm, 4.6×75mm), 60°C Detection wavelength: 264 nm Mobile phase A: 50 mM TEAAaq. Mobile phase B: MeOH Flow rate: 0.75 mL / min Gradient: 70-95%B (0-20min) 95%B (20-25min) 70%B (25-35min) LC / MS conditions: Equipment used: Ultra-high performance liquid chromatograph ACQUITY UPLC (waters) Quadrupole time-of-flight mass spectrometer SYNAPT-MS(waters) Column:YMC-Triart C8 1.9μm 2.1x50mm(YMC) Temperature: 50℃ Flow rate: 0.4mL / min Mobile phase: 10 mM ammonia water Mobile phase: MeOH Gradient: 70-95%B (8min) Detector 1: UV 264 nm Detector 2: Quadrupole time-of-flight mass spectrometer Ionization method: ESI+ Measurement range: 100-2000m / z

[0151] Example 1 4-(octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl (hereinafter referred to as "G1-suc-morT-OFF") Step 1: Production of 4-(octadecylamino)-4-oxobutanoic acid (hereinafter referred to as "G1-suc") Succinic anhydride (8.96g, 1.1eq) and triethylamine (17mL, 1.5eq.) were added to a solution of octadecane-1-amine (21.94g) in dichloromethane (500mL) and stirred at room temperature for 7 hours. The mixture was concentrated under reduced pressure, and 150mL of acetone was added to the residue and stirred for 16 hours. The precipitate was filtered by suction, washed with acetone (400mL), and then dried under reduced pressure at 30℃ for 3 hours to obtain G1-suc (29.1g, 96.6%) as a white powder.

[0152] Step 2: Preparation of 4-(octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl (hereinafter referred to as "G1-suc-morT-OFF") 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. Then, 1-((2R,6S)-6-(hydroxymethyl)-4-tritylmorpholin-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (hereinafter referred to as "morT-OH") (18 g, 1.0 eq.) and 4-(N,N-dimethylamino)pyridine (4.57 g) were added and the mixture was stirred for 1 hour on a 70°C water bath. After cooling to room temperature, 0.1 M aqueous sodium dihydrogen phosphate 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 and once with saturated saline diluted twice 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 distilled off and dried under reduced pressure to obtain 4-(octadecylamino)-4-oxobutanoate [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methyl (hereinafter referred to as "G1-suc-morT-ON") (white amorphous, 28.1 g, 89.5%). This was dissolved in 140 mL of dichloromethane, and 20 mL of 2,2,2-trifluoroethanol and 10.3 mL of triisopropylsilane were added while stirring on an ice bath, and after stirring for a while, 5.1 mL of trifluoroacetic acid was added dropwise. One hour after the end 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, purified by column chromatography on silica gel using a mixture of dichloromethane and methanol as the mobile phase, and dried under reduced pressure to obtain 19.89 g of G1-suc-morT-OFF as a powder. 1 H-NMR (CDCl 3):δ8.90(1H,bs);7.25(1H,d,J=1.6Hz);5.72(1H,dd,J=9.6Hz,2.6Hz);5.65(1H,m) ;4.14(2H,d,J=5.2Hz);3.98(1H,m);3.23(2H,dd,J=12.8Hz,7.0Hz);3.12(2H,dd,J =12Hz,2.6Hz);2.95(2H,dd,J=12.8Hz,1.8Hz);2.60~2.75(4H,m);2.47(2H,t,J=6. 8Hz);1.95(3H,d,J=1.6Hz);1.48(2H,m), 1.21~1.34(29H,m);0.88(3H,t,J=6.4Hz) ESI-MS(+):593.36(M+H)

[0153] 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") Step 1: Production of 4-((1,3-bis(stearoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (hereinafter referred to as "G2-suc") Dichloromethane (8mL) was added to 1g (1.60mmol) of 2-hydroxypropane-1,3-diyl distearate, followed by 176mg (1.76mmol) of succinic anhydride and 293mg (2.40mmol) of 4-(N,N-dimethylamino)pyridine, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, 1M aqueous sodium dihydrogen phosphate solution was added to the reaction solution, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was removed to obtain G2-suc (1.40g).

[0154] 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 (5.2mL) was added to G2-suc (900mg, 1.24mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (277mg, 1.45mmol), followed by morT-OH (500mg, 1.03mmol) and 4-(N,N-dimethylamino)pyridine (132mg, 1.09mmol), and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, 0.1M aqueous sodium dihydrogen phosphate was added to the reaction solution, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was removed by distillation. The resulting residue was purified by silica gel chromatography to obtain G2-suc-morT-ON (1.09g, 89%). 1 H-NMR (CDCl3 ):δ8.04(1H,s);7.17~7.51(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);1.38~1.44(2H,m);1.24(60H,m);0.87(6H,t,J=6.8Hz)

[0155] Process 3 Manufacturing of G2-suc-morT-OFF Dichloromethane (4.2 mL) was added to G2-suc-morT-ON and stirred at 0°C. Then, 127 μL (0.62 mmol) of triisopropylsilane and 64 μL (0.82 mmol) of trifluoroacetic acid were added at 0°C and stirred at room temperature for 1 hour. After the reaction was completed, a saturated aqueous solution of sodium bicarbonate was added to the reaction solution, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was removed by distillation. The resulting residue was purified by silica gel chromatography to obtain G2-suc-morT-OFF (373 mg, 95%). 1 H-NMR (CDCl 3 ):δ8.04(1H,bs);7.24(1H,s);5.70(1H,d,J=2Hz);5.21~5.26(1H,m);4.28~4.31(2H,m);4.13~4.17(4H,m);3.96~4.00(1H,m);3.11(1H, dd,J=12.4,2Hz);2.94(1H,dd,J=12.8,2.4Hz);2.57~2.65(6H,m);2.32(4H,t,J=7.6Hz);1.95(3H,s);1.25(60H,m);0.88(6H,t,J=7.6Hz)

[0156] 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]succinic acid (hereinafter referred to as "G3-suc-morT-OFF") 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 raw material, 4-((1,3-bis(oleoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (hereinafter referred to as "G3-suc") was produced in the same manner as in step 1 of Example 2. Then, G3-suc-morT-ON was produced in the same manner as in step 2 of Example 2. 1 H-NMR (CDCl 3 ):δ8.00(1H,s);7.17~7.51(15H,m);6.99(1H,s);6.09~6.12(1H,m);5.29~5.38(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 );1.97~2.02(8H,m);1.83(3H,s);1.57~1.61(2H,m);1.28(44H,m);0.89(6H,t,J=6.8Hz)

[0157] Process 2 Manufacturing of G3-suc-morT-OFF It was produced in the same manner as in step 3 of Example 2. 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);5.2 1~5.25(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)

[0158] Example 4 4-oxo-4-(4-stearoylpiperazin-1-yl)butanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl (hereinafter referred to as "G4-suc-morT-OFF") Step 1: Production of 4-oxo-4-(4-stearoylpiperazin-1-yl)butanoic acid (hereinafter referred to as "G4-suc") 1.68g (5.91mmol) of stearic acid, 1.13g (5.91mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 0.79g (5.91mmol) of 1-hydroxybenzotriazole were added to 26mL of tetrahydrofuran, then 1.45mL (10.7mmol) of triethylamine and 1g (5.37mmol) of tert-butyl piperazine-1-carboxylate were added, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, a saturated aqueous solution of sodium bicarbonate was added to the reaction solution, extracted with dichloromethane, dried with sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel chromatography to obtain 4-stearoylpiperazine-1-carboxylate tert-butyl (1.64g; 67%). 18mL of dichloromethane was added to this and stirred at 0°C, and 2.77mL (36.2mmol) of trifluoroacetic acid was added at 0°C, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, a saturated aqueous solution of sodium bicarbonate was added to the reaction solution, extracted with dichloromethane, dried with sodium sulfate, and the solvent was distilled off to obtain 1-(piperazin-1-yl)octadecan-1-one (1.30 g). 18 mL of dichloromethane was added to 1.3 g (3.70 mmol) of the crude product, followed by 0.41 g (4.10 mmol) of succinic anhydride and 0.77 mL (5.50 mmol) of triethylamine, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was distilled off, and acetone was added to the residue and the mixture was slurry washed at room temperature for 16 hours. The insoluble matter was collected by suction filtration, washed with acetone, and dried to obtain G4-suc (1.20 g).

[0159] Step 2: Preparation of 4-oxo-4-(4-stearoylpiperazin-1-yl)butanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl (hereinafter referred to as "G4-suc-morT-ON") Tetrahydrofuran (10 mL) was added to G4-suc (982 mg, 2.17 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (555 mg, 2.90 mmol), and the mixture was stirred at 70°C. Then, morT-OH (1 g, 2.07 mmol) and 4-(N,N-dimethylamino)pyridine (265 mg, 2.17 mmol) were added, and the mixture was stirred at 70°C for 30 minutes. After the reaction was completed, the mixture was cooled to room temperature, and 0.1 M aqueous sodium dihydrogen phosphate was added to the reaction solution, which was extracted with dichloromethane, dried over sodium sulfate, and the solvent was removed by distillation. The resulting residue was purified by silica gel chromatography to obtain G4-suc-morT-ON (1.68 g, 89%). 1 H-NMR (CDCl 3 ):δ8.00(1H,s);7.16~7.50(15H,m);6.97(1H,s);6.10(1H,d,J=8Hz);4.34~4.36(1H,m);4.04(2H,d,J=4.8Hz);3.57~3.64(4H,m);3.44~3.4 8(4H,m);3.32~3.34(1H,m);3.09~3.12(1H,m);2.60~2.64(4H,m);2.3 1(2H,t,J=7.6Hz);1.82(3H,s);1.23~1.42(32H,m);0.86(3H,t,J=6.8)

[0160] Process 3 Manufacturing of G4-suc-morT-OFF It was produced in the same manner as in step 3 of Example 2. 1 H-NMR (CDCl 3 ):δ8.32(1H,bs);7.23(1H,s);5.67~5.70(1H,m);4.12~4.19(2H,m);3.96~4.01(1H,m);3.47~3.67(8H,m)3.10~3.13(1H ,m);2.95~2.98(1H,m);2.60~2.72(4H,m);2.33(2H,t,J=7.2Hz);1.95(3H,s);1.25~1.31(32H,m);0.88(3H,t,J=7.6Hz)

[0161] Example 5 4-(Octadecylcarbamoyl)benzoic acid [(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)benzoic acid [(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 Example 2 using 4-(octadecylcarbamoyl)benzoic acid. 1 H-NMR (CDCl 3 ):δ8.24(1H,s);7.97(2H,d,J=8Hz);7.77(2H,d,J=8Hz);7.17~7.46(15H,m);6.95(1H,s);6.12~6.16(1H,m);4.49~4.51(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)

[0162] Step 2: Production of G5-tpa-morT-OFF It was produced in the same manner as in step 3 of Example 2. 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);6.14~6.17(1H,m);5.74~5.77(1H,m);4.40~4.45(2H,m) ;4.13~4.19(1H,m);3.45~3.50(2H,m);3.14~3.18(1H,m);3.05~3.08(1H,m);1.93(3H,s);1.26~1.41(34H,m);0.89(3H,t,J=7.6Hz)

[0163] Example 6 4-(4-(4-(octadecylcarbamoyl)benzoyl)piperazin-1-yl)-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl (hereinafter referred to as "G6-suc-morT-OFF"). Step 1: Production of 4-(4-(4-(octadecylcarbamoyl)benzoyl)piperazin-1-yl)-4-oxobutanoic acid (hereinafter referred to as "G6-suc") This was produced in the same manner as in step 1 of Example 4, except that 4-(octadecylcarbamoyl)benzoic acid was used instead of stearic acid.

[0164] Step 2: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl 4-[4-{4-(octadecylcarbamoyl)benzoyl}piperazin-1-yl]-4-oxobutanoate (hereinafter referred to as "G6-suc-morT-ON"). It was produced in the same manner as in step 2 of Example 2. 1 H-NMR (CDCl 3):δ8.08(1H,bs);7.81(2H,d,J=7.6Hz);7.16~7.50(17H,m);6.97(1H,s);6.08~6.10(1H,m);4.33~4.39(1H,m);4.02~4.0 4(2H,m);3.31~3.79(11H,m);3.08~3.11(1H,m);2.60~2.69(4H,m);1.81(3H,s);1.23~1.44(34H,m);0.86(3H,t,J=6.4Hz)

[0165] Process 3 Manufacturing of G6-suc-morT-OFF To 493 mg (0.47 mmol) of G6-suc-morT-ON, 4.6 mL of dichloromethane and 0.4 mL of 2,2,2-trifluoroethanol were added and stirred at 0°C. Then, 145 μL (0.70 mmol) of triisopropylsilane and 53 μL (0.70 mmol) of trifluoroacetic acid were added at 0°C and stirred at room temperature for 1 hour. After the reaction was completed, a saturated aqueous solution of sodium bicarbonate was added to the reaction solution, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was removed by distillation. The resulting residue was purified by silica gel chromatography to obtain G6-suc-morT-OFF (372 mg; 98%). 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(1H,s);6.08~6.11(1H,m);5.67~5.69(1H,m);4.10~4.15(2H,m);3.96~3. 99(1H,m);3.36~3.79(8H,m);3.08~3.11(1H,m);2.93~2.96(1H,m);2. 57~2.70(6H,m);1.92(3H,s);1.23~1.38(34H,m);0.86(3H,t,J=7.2Hz)

[0166] Example 7 3,4,5-tris(octadecyloxy)benzoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl (hereinafter referred to as "G7-morT-OFF") Step 1: Preparation of 3,4,5-tris(octadecyloxy)benzoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-trityl-morpholin-2-yl}methyl (hereinafter referred to as "G7-morT-ON"). This was prepared in the same manner as in Step 2 of Example 2 using 3,4,5-trioctadecoxybenzoic acid. 1 H-NMR (CDCl 3):δ7.90(1H,bs);7.12~7.45(17H,m);6.97(1H,s);6.12~6.14(1H,m);4.46~4.51(1H,m);4.28~4.32(1H,m);4.16~4.20(1H ,m);3.90~4.00(6H,m);3.37~3.40(1H,m);3.22~3.25(1H,m);1.78~1.82(5H,m);1.23~1.50(96H,m);0.86(9H,t,J=6.8Hz)

[0167] Step 2: Preparation of G7-morT-OFF It was produced in the same manner as in step 3 of Example 2. 1 H-NMR (CDCl 3 ):δ7.98(1H,bs);7.22(3H,m);5.69~5.72(1H,m);4.32~4.36(2H,m);4.08~4.12(1H,m);3.94~4.01(6H,m);3 .11~3.14(1H,m);3.02~3.05(1H,m);2.64~2.72(2H,m);1.90(3H,m);1.23~1.45(96H,m)0.86(9H,t,J=7.2Hz)

[0168] 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"). 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 the reaction was completed, 1 M aqueous sodium dihydrogen phosphate was added to the reaction solution, extracted with dichloromethane, dried over sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel chromatography to obtain 2-hydroxyethyl 3,4,5-trioctadecyloxybenzoate (882 mg; 56%). 1 H-NMR (CDCl 3):δ7.26(2H,s);4.45~4.47(2H,m);3.95~4.03(8H,m);1.25~1.52(96H,m);0.88(9H,t,J=7.2Hz)

[0169] Step 2: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl (2-[{3,4,5-tris(octadecyloxy)benzoyloxy}oxy]ethyl) succinate (hereinafter referred to as "G8-suc-morT-ON"). 4-oxo-4-(2-[{3,4,5-tris(octadecyloxy)benzoyl}oxy]ethoxy)butanoic acid (hereinafter referred to as "G8-suc") was obtained in a manner similar to that of step 1 of Example 2, and then G8-suc-morT-ON was obtained in a manner similar to that of step 2 of Example 2. 1 H-NMR (CDCl 3 ):δ7.87(1H,bs);7.12~7.43(17H,m);6.97(1H,s);6.07~6.10(1H,m);4.33~4.46(5H,m);3.91~4.07(8H,m);3.31 ~3.34(1H,m);3.07~3.10(1H,m);2.56~2.60(4H,m);1.68~1.80(5H,m);1.23~1.50(96H,m);0.86(9H,t,J=7.2Hz)

[0170] Step 3: Preparation of G8-suc-morT-OFF G8-suc-morT-OFF was obtained in the same manner as in step 3 of Example 2. 1 H-NMR (CDCl 3 ):δ7.96(1H,bs);7,23(3H,m);5.67~5.69(1H,m);4.40~4.47(5H,m);3.94~4.11(8H,m);3.09~3.12 (1H,m);2.89~2.92(1H,m);2.53~2.65(6H,m)1.90(3H,s);1.23~1.45(96H,m);0.86(9H,t,J=6.8Hz)

[0171] 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 (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-octadecyl octadecane-1-amine as a raw material, 4-(dioctadecylamino)-4-oxobutanoic acid (hereinafter referred to as "G9-suc") was produced in the same manner as in step 1 of Example 2. Then, G9-suc-morT-ON was produced in the same manner as in step 2 of Example 2. 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);1.82(3H,s);1.23~1.52(64H,m);0.85(6H,t,J=6.8Hz)

[0172] Step 2: Preparation of G9-suc-morT-OFF It was produced in the same manner as in step 3 of Example 2. 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)

[0173] 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 (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 (hereinafter referred to as "G10-suc-morT-ON") 9.4mL of tetrahydrofuran was added to 500mg (1.88mmol) of 2-tert-butoxycarbonylamino-3-phenyl-propanoic acid, followed by 652μL (3.77mmol) of N-ethyl-N-isopropyl-propan-2-amine, 46mg (0.38mmol) of 4-(N,N-dimethylamino)pyridine, 505mg (2.64mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, and octadecane-1-amine, and the mixture was stirred at room temperature for 5 hours. After the reaction was completed, 1M aqueous sodium dihydrogen phosphate was added to the reaction solution, extracted with dichloromethane, dried with sodium sulfate, and the solvent was distilled off. The resulting residue was purified by silica gel chromatography to obtain tert-butoxycarbonylamino-N-octadecyl-3-phenyl-propanamide (779mg, 80%). 1 H-NMR (CDCl 3 ):δ7.18~7.28(5H,m);5.57(1H,bs);5.06(1H,bs);4.20~4.26(1H,m);2.95~3.12(4H,m);1.40(9H,s);1.14~1.28(32H,m);0.86(3H,t,J=6.8Hz)

[0174] 15 mL of dichloromethane was added to 779 mg (1.51 mmol) of tert-butoxycarbonylamino-N-octadecyl-3-phenyl-propanamide, 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 the same reaction as in step 1 of Example 2 to produce G10-suc. Then, the same reaction as in step 2 of Example 2 was performed to produce G10-suc-morT-ON. 1 H-NMR (CDCl 3):δ8.08(1H,bs);7.16~7.32(20H,m);6.98(1H,s);6.29~6.31(1H,m); 6.10~6.12(1H,m);5.56~5.59(1H,m);4.51~4.57(1H,m);4.35~4.37(1 H,m);4.02(2H,d,J=5.6Hz);3.73~3.77(1H,m);2.92~3.33(6H,m);2.39~2.67(4H,m);1.84(3H,s);1.21~1.45(32H,m);0.88(3H,t,J=7.6Hz)

[0175] Process 2 Manufacturing of G10-suc-morT-OFF The target substance was obtained in the same manner as in step 3 of Example 2. 1 H-NMR (CDCl 3 ):δ8.33(1H,bs);7.16~7.32(6H,m);6.38~6.40(1H,m);5.67~5.71(2H,m);4.54~4.58(1H,m);4.08~4.16(3H,m) ;3.94~4.01(1H,m);2.91~3.17(5H,m);2.46~2.79(5H,m);1.94(3H,s);1.13~1.36(32H,m);0.88(3H,t,J=7.6Hz)

[0176] The chemical structural formulas of the compounds described in Examples 1 to 10 above are shown in Table 3 below. [Table 3]

[0177] Example 11: G1-suc-PMO[A Bz -A Bz ]-ON (Tetrabutylammonium chloride was used as a reaction promoter, reaction time (30 min)) [ka] G1-suc-morA-OFF 14.1mg (0.02mmol), N-ethylmorpholine 7.6μL (0.06mmol) and tetrabutylammonium chloride 16.7mg (0.06mmol) were dissolved in dichloromethane 200μL. 21.7mg (0.03mmol) of morA was dissolved in dichloromethane 200μL and shaken at 40℃ for 30 minutes. After 30 minutes, 160μL of 1% 1-methylpiperazine / dichloromethane solution was added to 40μL of the reaction solution to stop the reaction. This solution was further diluted 10 times with acetonitrile and analyzed by HPLC (raw material Rt: 13.20min, target product Rt: 19.68min, conversion yield 99.4%). ESI-TOF-MS(+) Measured value: 1391.91 (M+H), theoretical value: 1390.71. G1-suc-morA-OFF: ((2S,6R)-6-(6-benzamido-9H-purin-9-yl)morpholin-2-yl)methyl 4-(octadecylamino)-4-oxobutanoate G1-suc-morA-OFF was synthesized by using "N-(9-((2R,6S)-6-(hydroxymethyl-4-tritylmorpholin-2-yl)-9H-purin-6-yl)benzamide (morA-OH)" instead of "morT-OH" in step 2 of Example 1.

[0178] Example 12: G1-suc-PMO[A Bz -T]-ON (1-butyl-1-methylpyrrolidinium chloride was used as the reaction promoter, reaction time (60 min)) [ka] G1-suc-morA-OFF 10mg (0.0142mmol), morT 12.9mg (0.0212mmol), N-ethylmorpholine 5.38μL (0.0425mmol) and 1-butyl-1-methylpyrrolidinium chloride 7.55mg (0.0425mmol) were dissolved in dichloromethane 283μL and stirred at room temperature for 60 minutes. After 60 minutes, 1-methylpiperazine 4.71μL (0.0425mmol) was added to the reaction solution to stop the reaction. This solution was further diluted 60 times with acetonitrile and analyzed by HPLC (raw material Rt: 13.21min, target product Rt: 18.77min, conversion yield 99.7%). ESI-TOF-MS(+) Measured value: 1278.76 (M+H), Theoretical value: 1277.68

[0179] Example 13: G1-suc-PMO[A Bz -G CE,Pac -TTTC Bz -TT]-OFF (1-butyl-1-methylpyrrolidinium chloride was used as a reaction promoter) The following liquid a was fed at 0.05 mL / min and liquid b was fed at 0.2 mL / min, and the two liquids were mixed and then reacted in a 30 mL tube reactor at 40 ° C for 120 minutes. After the reaction, it was mixed with the following liquid c fed at 0.6 mL / min, and reacted in a 5 mL tube reactor at 40 ° C for 5.9 minutes. Then, it was mixed with the following liquid d fed at 0.85 mL / min and collected in a dropping funnel. The organic layer in the dropping funnel was fed at 0.8 mL / min, mixed with the following liquid e fed at 0.4 mL / min, and reacted in a 15.6 mL tube reactor at room temperature for 13 minutes. After the reaction, it was further mixed with the following liquid f fed at a flow rate of 1.5 mL / min and collected in a separating funnel. The collected organic layer was washed with the following liquid g and saturated sodium bicarbonate water, dried with sodium sulfate, and the solvent was distilled off. After that, it was dissolved in a small amount of dichloromethane, diisopropyl ether was added, and the resulting precipitate was filtered and dried under reduced pressure overnight to obtain G1-suc-PMO[A Bz -G CE,Pac -TTTC Bz-TT]-OFF (white solid, 2.53 g) was obtained. A small amount was dissolved in dichloromethane, diluted with a 1:1 mixed solvent of methanol and acetonitrile, and analyzed by HPLC (raw material Rt: 13.19 min, target product Rt: 12.84 min, conversion yield 96.7%). ESI-TOF-MS(+) Actual value: 3317.39, Theoretical value: 3317.32

[0180] Solution a: 3.17 g (5.2 mmol) of morT was dissolved in 5 mL of dichloromethane. Solution b: G1-suc-PMO[A Bz -G CE,Pac -TTTC Bz 2.94 g (0.984 mmol) of -T]-OFF, 373 μL (2.95 mmol) of N-ethylmorpholine, and 2.95 mL (2.95 mmol) of a dichloromethane solution (1 M) of 1-butyl-1-methylpyrrolidinium chloride were dissolved in a mixed solution of 1.36 mL of 1,3-dimethyl-2-imidazolidinone and 6.09 mL of dichloromethane. Solution c: 3.33 mL (30 mmol) of 1-methylpiperazine was dissolved in 96.7 mL of dichloromethane. Liquid d: Aqueous solution of sodium dihydrogen phosphate (1M) and saturated saline were mixed in a 1:1 ratio, and 5% ethanol was added by volume. Solution e: 4 mL of trifluoroacetic acid, 1 mL of ethanol, 1 mL of triethylamine, 30 mL of trifluoroethanol, and 64 mL of dichloromethane were mixed. Liquid f: Saturated saline and water mixed in a 1:1 ratio. Solution g: 1% trifluoroacetic acid aqueous solution and saturated saline were mixed in a 1:1 ratio, and 5% ethanol was added by volume.

[0181] Example 14: The reaction promoter (1-butyl-1-methylpyrrolidinium chloride: BMPC) and solvent (1,3-dimethyl-2-imidazolidinone / dichloromethane) were used. G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -ABz -TC Bz -C Bz -A Bz -G CE,Pac -TA Bz ]-ON (1-butyl-1-methylpyrrolidinium chloride was used as the reaction promoter, 1,3-dimethyl-2-imidazolidinone / dichloromethane was used as the solvent, and the reaction time was 120 minutes) G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TC Bz -C Bz -A Bz -G CE,Pac -T]-OFF 1.26g (0.213mmol), morA 230mg (0.319mmol), N-ethylmorpholine 81μL (0.638mmol) and 1-butyl-1-methylpyrrolidinium chloride 113mg (0.638mmol) were dissolved in 1,3-dimethyl-2-imidazolidinone / dichloromethane solution 7.1mL and stirred at 40℃ for 120 minutes. After 120 minutes, 1-methylpiperazine 71μL (0.638mmol) was added to the reaction solution to stop the reaction. This solution was further diluted 60 times with 80% acetonitrile aqueous solution and analyzed by HPLC (raw material Rt: 15.16min, target product Rt: 18.49, 18.72min, conversion yield 97.8%).

[0182] G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TC Bz -C Bz -A Bz -G CE,Pac -TA Bz -T]-ON (1-butyl-1-methylpyrrolidinium chloride was used as the reaction promoter, 1,3-dimethyl-2-imidazolidinone / dichloromethane was used as the solvent, and the reaction time was 120 minutes) G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TCBz -C Bz -A Bz -G CE,Pac -TA Bz ]-OFF 1.17g (0.209mmol), morT 191mg (0.314mmol), N-ethylmorpholine 79μL (0.628mmol) and 1-butyl-1-methylpyrrolidinium chloride 186mg (1.05mmol) were dissolved in 1,3-dimethyl-2-imidazolidinone / dichloromethane solution 7.0mL and stirred at 40℃ for 120 minutes. After 120 minutes, 1-methylpiperazine 70μL (0.628mmol) was added to the reaction solution to stop the reaction. This solution was further diluted 60 times with 80% acetonitrile aqueous solution and analyzed by HPLC (raw material Rt: 15.18min, target product Rt: 18.17min, conversion yield 99.6%).

[0183] G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TC Bz -C Bz -A Bz -G CE,Pac -TA Bz -TA Bz ]-ON (1-butyl-1-methylpyrrolidinium chloride was used as the reaction promoter, 1,3-dimethyl-2-imidazolidinone / dichloromethane was used as the solvent, and the reaction time was 120 minutes) G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TC Bz -C Bz -A Bz -G CE,Pac -TA Bz-T]-OFF 1.08g (0.182mmol), morA 198mg (0.274mmol), N-ethylmorpholine 69μL (0.547mmol) and 1-butyl-1-methylpyrrolidinium chloride 162mg (0.912mmol) were dissolved in 1,3-dimethyl-2-imidazolidinone / dichloromethane solution 6.1mL and stirred at 40℃ for 120 minutes. After 120 minutes, 1-methylpiperazine 61μL (0.547mmol) was added to the reaction solution to stop the reaction. This solution was further diluted 60 times with 80% acetonitrile aqueous solution and analyzed by HPLC (raw material Rt: 15.02min, target product Rt: 18.12, 18.32min, conversion yield 99.4%). ESI-TOF-MS(+) Actual value: 6362.332, Theoretical value: 6362.320

[0184] Example 15: A reaction promoter (1-butyl-1-methylpyrrolidinium chloride: BMPC) and a solvent (dichloromethane) were used. G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TC Bz -C Bz -A Bz -G CE,Pac -TA Bz ]-ON (1-butyl-1-methylpyrrolidinium chloride was used as the reaction promoter, dichloromethane was used as the solvent, and the reaction time was 120 minutes) G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TC Bz -C Bz -A Bz -G CE,Pac-T]-OFF 2.14g (0.361mmol), morA 391mg (0.542mmol), N-ethylmorpholine 137μL (1.08mmol) and 1-butyl-1-methylpyrrolidinium chloride 193mg (1.08mmol) were dissolved in dichloromethane 12mL and stirred at 40℃ for 120 minutes. After 120 minutes, 1-methylpiperazine 120μL (1.08mmol) was added to the reaction solution to stop the reaction. This solution was further diluted 60 times with 80% acetonitrile aqueous solution and analyzed by HPLC (raw material Rt: 15.16min, target product Rt: 18.49, 18.72min, conversion yield 97.8%).

[0185] G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TC Bz -C Bz -A Bz -G CE,Pac -TA Bz -T]-ON (1-butyl-1-methylpyrrolidinium chloride was used as the reaction promoter, dichloromethane was used as the solvent, and the reaction time was 120 minutes) G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TC Bz -C Bz -A Bz -G CE,Pac -TA Bz ]-OFF 2.0g (0.357mmol), morT 326mg (0.536mmol), N-ethylmorpholine 136μL (1.07mmol) and 1-butyl-1-methylpyrrolidinium chloride 318mg (1.79mmol) were dissolved in dichloromethane 11.9mL and stirred at 40℃ for 120 minutes. After 120 minutes, 1-methylpiperazine 119μL (1.07mmol) was added to the reaction solution to stop the reaction. This solution was further diluted 60 times with 80% acetonitrile aqueous solution and analyzed by HPLC (raw material Rt: 15.18min, target product Rt: 18.17min, conversion yield 99.7%).

[0186] G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TC Bz -C Bz -A Bz -G CE,Pac -TA Bz -TA Bz ]-ON (1-butyl-1-methylpyrrolidinium chloride was used as the reaction promoter, dichloromethane was used as the solvent, and the reaction time was 120 minutes) G1-suc-PMO[TG CE,Pac -G CE,Pac -G CE,Pac -A Bz -TC Bz -C Bz -A Bz -G CE,Pac -TA Bz -T]-OFF 1.90g (0.321mmol), morA 348mg (0.481mmol), N-ethylmorpholine 122μL (0.962mmol) and 1-butyl-1-methylpyrrolidinium chloride 285mg (1.60mmol) were dissolved in dichloromethane 10.7mL and stirred at 40℃ for 120 minutes. After 120 minutes, 1-methylpiperazine 107μL (0.962mmol) was added to the reaction solution to stop the reaction. This solution was further diluted 60 times with 80% acetonitrile aqueous solution and analyzed by HPLC (raw material Rt: 15.02min, target product Rt: 18.12, 18.32min, conversion yield 97.0%).

Claims

1. General formula [A-1]: 【Chemistry 1】 [In the formula, B P represents an optionally protected nucleic acid base, and the nucleic acid base is adenine, guanine, hypoxanthine, cytosine, thymine, uracil, or a modified base thereof, and the modified base is pseudouracil, 3-methyluracil, dihydrouracil, 5-alkylcytosine, 5-alkyluracil, 5-halouracil, 6-azapyrimidine, 6-alkylpyrimidine, 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; W represents an oxygen atom; X's are the same or different and each is di(C 1-6 alkyl)amino; G has the following formula: 【Chemistry 2】 (In the formula, * represents the bonding position with T.) selected from the group consisting of: T is a single bond; n represents an integer of 1 to 50. A compound [A-1] represented by the formula: General formula [B-1]: 【Chemistry 3】 [In the formula, B P , W, and X are as defined above; D is a halogen; p represents an integer from 1 to 10; Q 1 represents a group removable under acidic conditions, and the group removable under acidic conditions is selected from the group consisting of trityl, monomethoxytrityl, tert-butyldimethylsilyl, and dimethoxytrityl.] The compound [B-1] represented by the general formula [C-1] is subjected to a condensation reaction. 【Chemistry 4】 [In the formula, B P , Q 1 , W, X, G, T, n and p are as defined above.] The method for producing the compound [C-1] represented by the formula (I) is characterized in that the method is carried out in the presence of at least one reaction promoter selected from the group consisting of quaternary ammonium salts, quaternary piperidinium salts, and quaternary pyrrolidinium salts.

2. The method according to claim 1, wherein the reaction promoter is at least one selected from the group consisting of quaternary ammonium salts and quaternary pyrrolidinium salts.

3. The reaction promoter is Tetra C １－１８ Alkyl ammonium chloride and 1-C １－１８ Alkyl-1-C １－１８ 2. The method according to claim 1, wherein the alkyl group is at least one selected from the group consisting of alkylpyrrolidinium chlorides.

4. The reaction accelerator is Tetrabutylammonium chloride, tetrabutylammonium bromide, dodecyltrimethylammonium chloride, Trioctylmethylammonium chloride, Choline chloride, N,N,N-trimethylbutane-1-amino chloride, and 1-Butyl-1-methylpyrrolidinium chloride The method of claim 1, wherein the at least one selected from the group consisting of:

5. 2. The method according to claim 1, wherein the reaction promoter is at least one selected from the group consisting of tetrabutylammonium chloride and 1-butyl-1-methylpyrrolidinium chloride.

6. Furthermore, by adding a solution containing an acid to a reaction mixture containing the compound [C-1] produced by the method of claim 1, Q １ The elimination reactions of the above are carried out continuously in the same system to obtain a compound represented by the following general formula [E-1]: 【Chemistry 5】 [In the formula, n, p, B Ｐ , G, T, W and X are as defined above.] 2. The method of claim 1 , comprising forming a compound represented by

7. in the presence of at least one reaction promoter selected from the group consisting of quaternary ammonium salts, quaternary piperidinium salts, and quaternary pyrrolidinium salts; General formula [A-1-1]: 【Chemistry 6】 [In the formula, B Ｐ , Q １ , W, X, G and T are as defined above; n is 1 to 25. From the compound Q １ By detaching General formula [A-1]: 【Chemistry 7】 [In the formula, B Ｐ , W, X, G, T, and n are as defined above.] and forming a compound represented by the general formula [A-1]. General formula [B-1]: 【Chemistry 8】 [In the formula, B Ｐ , Q １ , W, X, D and p are as defined above.] By reacting with the compound General formula [C-1]: 【Chemistry 9】 [In the formula, n, p, B Ｐ , Q １ , W, X, G, and T are as defined above.] The method according to any one of claims 1 to 6, for preparing a compound of formula:

8. The reaction accelerator is Tetramethylammonium Chloride Tetraethylammonium chloride, Tetrapropylammonium chloride, Tetrabutylammonium chloride, tetrabutylammonium bromide, dodecyltrimethylammonium chloride, Trioctylmethylammonium chloride, N,N,N-trimethylbutane-1-amino chloride, and 1-Butyl-1-methylpyrrolidinium chloride and The amount of the reaction accelerator is in the range of 1 to 100 times the molar amount per 1 mole of the compound of the general formula [A-1]. The method of claim 7.

9. General formula [A-1-1]: 【Chemistry 10】 [In the formula, Q １ is trityl, monomethoxytrityl, or dimethoxytrityl, and n, B Ｐ , W, X, G, and T are as defined above.] From the compound In the presence of trifluoroacetic acid and 2,2,2-trifluoroethanol, and optionally triisopropylsilane or ethanol, Q １ including removing 9. The method according to claim 7 or 8.

10. A solution containing the compound of the general formula [A-1] and a solution containing the compound of the general formula [B-1] are fed into a flow reactor to form a compound of the general formula [C-1], and optionally A solution containing the compound of the formula [C-1] and a solution containing an acid are supplied to a flow reactor, １ to form a compound of the general formula [E-1], The method according to any one of claims 7 to 9.

11. A solution containing the compound of the general formula [A-1-1] and a solution containing an acid are supplied to a flow reactor, １ to form a compound of the general formula [A-1], and supplying a solution containing the compound of the general formula [A-1] and a solution containing the compound of the general formula [B-1] to a next flow reactor to form a compound of the general formula [C-1]; The method according to any one of claims 7 to 10.

12. General formula [A-1-2]: 【Chemistry 11】 [In the formula, B Ｐ is an optionally protected nucleic acid base, the nucleic acid base being adenine, guanine, hypoxanthine, cytosine, thymine, uracil, or a modified base thereof, the modified base being pseudouracil, 3-methyluracil, dihydrouracil, 5-alkylcytosine, 5-alkyluracil, 5-halouracil, 6-azapyrimidine, 6-alkylpyrimidine, 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; Q ２ is H or a group removable under acidic conditions, said group removable under acidic conditions being selected from the group consisting of trityl, monomethoxytrityl, tert-butyldimethylsilyl and dimethoxytrityl; W is an oxygen atom; X is Ji (C １－６ alkyl)amino; G is of the formula: 【Chemistry 12】 [In the formula, * represents the bonding position with T; T is a single bond; n is 1 to 25. Compound.

13. The protected nucleobase is a nucleobase in which the amino group or hydroxyl group of the nucleobase is protected with a protecting group, the protecting groups of the amino group being each independently selected from the group consisting of benzoyl, 4-methoxybenzoyl, acetyl, propionyl, butyryl, isobutyryl, phenylacetyl, phenoxyacetyl, 4-tert-butylphenoxyacetyl, 4-isopropylphenoxyacetyl, and (dimethylamino)methylene, and the protecting groups of the hydroxyl group being each independently selected from the group consisting of 2-cyanoethyl, 4-nitrophenethyl, phenylsulfonylethyl, methylamino ...

13. The method of any one of claims 1 to 11 or the compound of claim 12, wherein the aryl group is selected from the group consisting of arylsulfonylethyl, trimethylsilylethyl, phenyl optionally substituted at any substitutable position with 1 to 5 electron-withdrawing groups, diphenylcarbamoyl, dimethylcarbamoyl, diethylcarbamoyl, methylphenylcarbamoyl, 1-pyrrolidinylcarbamoyl, morpholinocarbamoyl, 4-(tert-butylcarboxy)benzyl, 4-[(dimethylamino)carboxy]benzyl, and 4-(phenylcarboxy)benzyl.

14. A reaction promoter for oligonucleic acid synthesis, comprising at least one active ingredient selected from the group consisting of quaternary ammonium salts, quaternary piperidinium salts and quaternary pyrrolidinium salts.

15. 15. The reaction promoter for oligonucleic acid synthesis according to claim 14, comprising at least one selected from the group consisting of quaternary ammonium salts and quaternary pyrrolidinium salts as an active ingredient.

16. The active ingredient is Tetra C １－１８ Alkyl ammonium chloride and 1-C １－１８ Alkyl-1-C １－１８ The reaction promoter for oligonucleic acid synthesis according to claim 14, comprising at least one selected from alkylpyrrolidinium chlorides.

17. As an active ingredient, Tetramethylammonium chloride, Tetraethylammonium chloride, Tetrapropylammonium chloride, Tetrabutylammonium chloride, tetrabutylammonium bromide, dodecyltrimethylammonium chloride, Trioctylmethylammonium chloride, N,N,N-trimethylbutane-1-amino chloride, and 1-Butyl-1-methylpyrrolidinium chloride The reaction promoter for oligonucleic acid synthesis according to claim 14, comprising at least one selected from the group consisting of:

18. The reaction promoter for oligonucleic acid synthesis according to claim 14, comprising at least one selected from the group consisting of tetrabutylammonium chloride and 1-butyl-1-methylpyrrolidinium chloride as an active ingredient.

19. The reaction promoter for oligonucleic acid synthesis according to any one of claims 14 to 18, for use in the method according to any one of claims 1 to 11.

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