Method for producing oligonucleic acid compound

Abstract

To provide a novel preparation method that can shorten the preparation period of an oligonucleic acid compound.SOLUTION: The present invention provides a method for producing a compound [C] represented, e.g., by the general formula [C-1] in the figure, by subjecting a compound [A] having a hydroxyl group or a primary or secondary amino group and a compound [B] having a phosphorous atom-containing substituent to a condensation reaction. The method is implemented in the presence of at least one reaction accelerator 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.SELECTED DRAWING: None

Description

[Technical Field]

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

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

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

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

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

[0009] The present inventors have discovered that phosphorus bonds can be efficiently formed 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 that can be removed under basic conditions 1-6 Alkyl)amino, di(amino-C substituted with a group that can be removed under basic conditions 1-6 alkyl)amino or the following general formula [2]: [ka] (In the formula, * indicates the bond position with P; a represents an integer of 0 to 2; E is CH2, CH-A 1 or NA 2 represents; A 1 is C 1-6 Alkyl, mono(C 1-6 Alkyl)amino-C 1-6 Alkyl, di(C 1-6 Alkyl)amino-C 1-6 Alkyl, Tri(C 1-6 Alkyl)ammonio-C 1-6 Alkyl, amino substituted with a group which can be removed under basic conditions, mono(C 1-6 alkyl)amino, di(C 1-6 alkyl)amino, tri(C 1-6 alkyl)ammonio, amino substituted with amidino substituted with a group that can be removed under basic conditions, or the following general formula [3]: [ka] (In the formula, * indicates the bond position with E; b represents an integer of 0 to 2; c represents 0 or 1; R 11 is C1-6 represents alkyl; M represents CH, an oxygen atom, a sulfur atom, or N- (a group that can be removed under basic conditions). represents a substituent represented by the formula (hereinafter referred to as "substituent [3]"); and A 2 is C 1-6 Alkyl, mono(C 1-6 Alkyl)amino-C 1-6 Alkyl, di(C 1-6 Alkyl)amino-C 1-6 Alkyl, Tri(C 1-6 Alkyl)ammonio-C 1-6 represents alkyl, a group that can be removed under basic conditions, aryl, or heteroaryl. (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 form 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. [Effects of the Invention]

[0011] Oligonucleic acid compounds are compounds having a structure in which two or more nucleoside units are linked via phosphorus bonds. To produce an oligonucleic acid compound, it is necessary to perform condensation reactions multiple times to form phosphorus bonds between adjacent nucleoside units. According to the present invention, phosphorus bonds can be formed efficiently, which is expected to result in a reduction in the time required to produce an oligonucleic acid compound. [Brief explanation of the drawings]

[0012] [Figure 1] Figure 1 shows 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 INVENTION

[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 and compound [B] having a substituent [1] to a condensation reaction, 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] Examples of the compound [A] that can be used in this production method include compounds having a hydroxyl group or a primary or secondary amino group.

[0015] A 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 bond position with the 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 substituents, (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 a 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 atom, a long-chain alkyl optionally substituted with 1 to 13 halogen atoms, or a long-chain alkyloxy optionally substituted with 1 to 13 halogen atoms; R 8e teeth, (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; and R 8f teeth, (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) represents a long-chain alkenyl-carbonyl. (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, * indicates the bonding position with Z; ** indicates the bond position with the oxygen atom; and L 1 is optionally substituted C 2-10 Alkylene or optionally substituted C 6-10 represents arylene.) (hereinafter referred to as "substituent

[10] "). represents a substituent represented by the formula (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 with 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. represents the bonding position to a substituent represented by (hereinafter referred to as "substituent [6]"); **teeth, (1) the bonding position of the phosphorus bond to the 3'-position oxygen atom or 3'-position nitrogen atom of the adjacent nucleoside unit; (2) The bond position with the hydrogen atom or (3) represents the bonding 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 represent a hydrogen atom or C 1-6 represents alkyl or R 4b1 and R 4b2 and 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 alkyl.) represents.] Examples of nucleoside units include those represented by the following formula (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 has the same meaning as 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 bond position with the hydrogen atom or (3) represents the bonding position to the substituent [6]; and **teeth, (1) the bonding position of the phosphorus bond to the 3'-position oxygen atom or 3'-position nitrogen atom of the adjacent nucleoside unit; (2) The bond position with the hydrogen atom or (3) Represents the bonding position with the substituent [6]. Examples of nucleoside units include those represented by the following formulae (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, those represented by the following general formula [5]: [ka] [In the formula, X has the same meaning as above; One of * and ** represents the bonding position to the 3'-position oxygen atom or 3'-position nitrogen atom of a nucleoside unit, and the other represents the bonding position to the 5'-position oxygen atom of a nucleoside unit different from the nucleoside unit; and W represents a lone pair, an oxygen atom, or a sulfur atom. (hereinafter referred to as "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 bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond, or (2) the bonding position with the 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 bond between each nucleoside unit constituting compound [A] may be the same or different, and may be, for example, phosphorus bond [5]. 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 specific embodiments of the compound [A] include, for example, compounds 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 of 1 to 50. (hereinafter referred to as "compound [A-1]") represented by the following formula: 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 specific embodiments of the compound [A] include, for example, compounds 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 that can be removed 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(C alkyl)amino; G is a compound having the formula: [ka] [In the formula, * represents the bonding position to T; T is a single bond; n is 1 to 25. Examples of the compounds include:

[0022] (A-2) Compound [A] composed of one to several 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 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 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 bonding position between the nucleoside unit and the oxygen atom at the 5' position of a nucleoside unit different from the nucleoside unit. More specific embodiments of the compound [A] include, for example, compounds 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]") represented by the following formula:

[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 bonding position with T.]

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

[0026] A specific embodiment of compound [B] is, for example, a compound containing one to several 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 has the same meaning as 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 with the substituent [1] or (3) represents the bond position to a group that can be removed under acidic conditions; ****teeth, (1) the bonding position of the phosphorus bond to the 3'-position oxygen atom or 3'-position nitrogen atom of the adjacent nucleoside unit; (2) the bonding position with the substituent [1] or (3) Represents the bonding position to a group that can be removed under acidic conditions.] Examples of nucleoside units include those represented by the following formula (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 has the same meaning as 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 with the substituent [1] or (3) represents the bond position to a group that can be removed under acidic conditions; and ****teeth, (1) the bonding position of the phosphorus bond to the 3'-position oxygen atom or 3'-position nitrogen atom of the adjacent nucleoside unit; (2) the bonding position with the substituent [1] or (3) Represents the bonding 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 bond between each nucleoside unit constituting compound [B] may be the same or different, and may be, for example, phosphorus bond [5]. 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.

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

[0031] (B-1) Compound [B] composed of one to several nucleoside units [4h] In the nucleoside unit represented by the general formula [4h], ***but, (1) The bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond, or (2) a bond site with 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 nucleoside unit on the 3'-terminal side is substituted with a group that can be removed under acidic conditions. In this case, the phosphorus bond between each nucleoside unit constituting compound [B] may be the same or different, and may be, for example, 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 nitrogen 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'-terminal side 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 substituent containing a phosphorus atom represented by the following formula: More specific embodiments of the compound [B] include, for example, compounds 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]") represented by the following formula: It should be noted that 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 listed in Table 1 below.

[0034] [Table 1]

[0035] (B-2) Compound [B] composed of one to several nucleoside units selected from the group consisting of nucleoside unit [4e], nucleoside unit [4f] and nucleoside unit [4g]. In the nucleoside units represented by the general formulae [4e], [4f] and [4g], ***but, (1) 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 bond between the oxygen atom at the 5' position of the adjacent nucleoside unit and the phosphorus bond, or (2) The bond position to 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 bond between each nucleoside unit constituting compound [B] may be the same or different, and may be, for example, phosphorus bond [5]. 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. The oxygen atom at the 3'-position of the nucleoside unit on the 3'-terminal side 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 substituent containing a phosphorus atom represented by the following formula: A more specific embodiment of the compound [B] is, for example, a 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]") represented by the following formula:

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

[0038] [Table 2]

[0039] (C) Compound [C] Examples of the compound [C] include compounds that can be produced by subjecting the compound [A] and the compound [B] to a condensation reaction.

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

[0041] (C-1) Compound [C] composed of one to several nucleoside units [4d] and one to several nucleoside units [4h] Specific embodiments of the compound [C] include, for example, 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]") represented by the following formula: As will be 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 with the oxygen atom at the 5' position of a nucleoside unit different from the nucleoside unit.

[0042] (C-2) Compound [C] composed of one to several nucleoside units selected from the group consisting of nucleoside unit [4a], nucleoside unit [4b] and nucleoside unit [4c], and one to several 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, a 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]") represented by the following formula: As will be 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 those represented by 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.] Examples include 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 (referred to as "compound [D-2]") can be converted into a compound represented by the formula:

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

[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 will be 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 this 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 needed. 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 this production method is, for example, suitably in 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 accelerator is used. The "reaction accelerator" 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. 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 can be used. 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 can be used. Even more preferably, at least one selected from the group consisting of quaternary ammonium salts, quaternary imidazolium salts, quaternary pyridinium salts, and quaternary pyrrolidinium salts can be used. Examples of quaternary ammonium salts that can be used in this 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 salts that can be used in this production method include, for example, tetra C 1-18 Alkylammonium salts (e.g., tetra C 1-18 Alkylammonium chloride), Tri C 1-18 Alkyl (hydroxy C 1-18 alkyl)ammonium salts, etc. (wherein C 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. 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 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 tricyanomethanide, 1-butyl-3-methylimidazolium 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 methyl sulfate, 1-ethyl-3-methylimidazolium diethyl phosphate, 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, and 1-methylimidazole, and preferably 1-ethyl-3-methylimidazolium chloride and 1-methyl-3-N-octylimidazolium chloride. The quaternary imidazolium salts that can be used in this production method include, 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 salts that can be used in this production method include, for example, tetra C 1-18 Alkylphosphonium salts, triC 1-18Alkyl (hydroxy C 1-18 alkyl)phosphonium salts (wherein C 1-18 The alkyls may be the same or different), preferably trihexyltetradecylphosphonium chloride. The quaternary pyridinium salts that can be used in this production method include, 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 salts that can be used in this production method include, 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, preferred reaction accelerators that can be used in the present production method include tetra C 1-18 Alkylammonium chloride, 1-C 1-18 Alkyl-1-C 1-18 Alkylpyrrolidinium chloride, 1-C 1-18 Alkyl-3-C 1-18 Examples include alkylimidazolium chlorides, 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 this production method is preferably within a range of 1 to 100 times, more preferably 1 to 50 times, and even more preferably 1.5 to 20 times, the molar ratio of which is based on 1 mole of compound [A]. The reaction temperature is suitably within the range of, for example, -78°C to 130°C, 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 suitably within the range of, for example, 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 to either a batch method or a flow method. Furthermore, this production method can also be applied to known methods for producing oligonucleic acid compounds, such as solid-phase methods, liquid-phase methods, and liquid-phase methods 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, the compound [A] can be supported on a solid support at the 3'-position oxygen atom of the 3'-terminal nucleoside unit or at the 5'-position oxygen atom of the 5'-terminal nucleoside unit of compound [A].

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

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

[0052] Hereinafter, the compound [C-1] and the compound [C-2] will be described in detail 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 this production method is not particularly limited as long as it is a solvent commonly used in the relevant technical field, and a single solvent may be used, or two or more solvents may be mixed. Examples 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. Two or more of these solvents may be mixed and used. In addition, examples of solvents that can be used in the present production method include polar solvents and halogenated solvents. Examples of polar solvents that can be used in this production method include dimethylacetamide, dimethyl sulfoxide, dimethylformamide, sulfolane, N-methylpiperidone, 1,3-dimethyl-2-imidazolidinone, N,N'-dimethylpropyleneurea, and mixtures thereof, of which dimethylacetamide, dimethyl sulfoxide, N-methylpiperidone, 1,3-dimethyl-2-imidazolidinone, and N,N'-dimethylpropyleneurea are preferred. Examples of halogen-containing solvents that can be used in this 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 proportion of the polar solvent in the mixed solvent of a polar solvent and a halogenated solvent that can be used in this production method has a lower limit of 1.0 wt%, preferably 2.0 wt%, more preferably 3.0 wt%, and even more preferably 5.0 wt%. The upper limit is 90 wt%, preferably 75 wt%, more preferably 50 wt%, even more preferably 40 wt%, and particularly preferably 30 wt%. Furthermore, these upper and lower limits can be used in appropriate combination; for example, a range of 1% to 90% is appropriate, a range of 5% to 75% is preferable, and a range of 5% to 50% is particularly preferable. In this production method, a base may be used as needed. 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 this production method is, for example, suitably in 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 needed. 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 this production method is, for example, suitably in a molar ratio of 0.2 to 6.0 times, preferably 0.4 to 3.0 times, and more preferably 1.0 to 2.5 times, per mole of compound [A]. The reaction temperature is suitably within the range of, for example, -78°C to 130°C, 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 suitably within the range of, for example, 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, the 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 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), the condensation reaction can be carried out, for example, 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 supply channels into a flow reactor or reaction channel, 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 for uninterrupted operation. Examples of means for supplying the solution containing compound [A-1] and the solution containing compound [B-1] to the supply flow path include pumps typically used in this field for supplying liquids, specifically, for example, syringe pumps, plunger pumps, diaphragm pumps, and gear pumps. Examples of the flow reactor include a microreactor and an in-line mixer such as a static mixer. As 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, for example, a multi-stage collision type micromixer can be mentioned. 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, or 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 may be selected appropriately, for example, from a range of 0.1 mm to 1.0 mm, and is preferably selected 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 needed. 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, the 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 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), the condensation reaction can be carried out, for example, by (1) stirring compound [A-2] and compound [B-2] in a reaction solvent in a suitable reaction vessel, or (2) supplying a solution containing compound [A-2] and a solution containing compound [B-2] independently through supply channels into a flow reactor or reaction channel, and mixing the solutions in the flow reactor or the like. Furthermore, after the condensation reaction, (1) the compound [C-2] can be obtained by purifying the reaction mixture using a column, or (2) adding an appropriate solvent to the reaction mixture, filtering the resulting precipitate, and washing it 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 all or 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], the compound can be produced by the same method 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 compounds [C-1] and [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] into 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 support. Furthermore, when G is a silicon substituent in the compounds [C-1] and [C-2], the target compounds 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 Desorption method 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 a case, a new compound is produced by selectively eliminating the group capable of being eliminated under acidic conditions in the molecule, and the condensation reaction described above in "(E) Method for producing compound [C]" can be carried out to produce compound [C] having one more nucleoside unit.

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

[0066] (G-1) Q in the molecule of compound [C-1] 1 Desorption method By reacting the compound [C-1] with an acid, Q substituted on the nitrogen atom at the 3'-position of the nucleoside unit on the 3'-terminal side of the compound [C-1] is obtained. 1 Q in the molecule of compound [C-1] can be eliminated. 1 By eliminating 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 on the 5'-position oxygen atom of the 5'-terminal nucleoside of the compound [C-1-1] 1 Before the elimination of the aryl group, the phosphorus atom on the phosphorus bond formed in the condensation reaction can 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 group [7] and Z is a solid 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 commonly used in the relevant technical field, and a single solvent may be used, or two or more solvents may be mixed. Examples 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. Two or more of these solvents may be mixed and used. 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 halogenated solvent can be used. The lower limit of the proportion of the polar solvent in the mixed solvent of a polar solvent and a halogenated solvent is 1.0 wt%, preferably 2.0 wt%, more preferably 3.0 wt%, and even more preferably 5.0 wt%. The upper limit is 90 wt%, preferably 75 wt%, more preferably 50 wt%, even more preferably 40 wt%, and particularly preferably 30 wt%. Furthermore, these upper and lower limits can be used in appropriate combination; for example, a range of 1% to 50% is appropriate, a range of 1% to 40% is preferable, and a 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 acid that can be used in this elimination reaction is suitably within the range of, for example, 1 to 500 times, and preferably 2 to 200 times, the molar ratio of the acid to 1 mole of the compound [C-1]. The acid that can be used in this elimination reaction is suitably diluted with an appropriate 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 as needed. 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 of 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), the elimination reaction can be carried out, for example, 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 halogenated solvent can be used. The lower limit of the proportion of the polar solvent in the mixed solvent of a polar solvent and a halogenated solvent is 1.0 wt%, preferably 2.0 wt%, more preferably 3.0 wt%, and even more preferably 5.0 wt%. The upper limit is 90 wt%, preferably 75 wt%, more preferably 50 wt%, even more preferably 40 wt%, and particularly preferably 30 wt%. Furthermore, these upper and lower limits can be used in appropriate combination, and the proportion of the polar solvent in the mixed solvent of a polar solvent and a halogenated solvent is, for example, suitably in the range of 1% to 50%, preferably 1% to 40%, and particularly preferably 1% to 30%. Examples of the "acid" that can be used in this elimination reaction include the same as those described above. When using these acids, they may be used in combination with a base (e.g., triethylamine) to adjust the acidity. The amount of acid that can be used in this elimination reaction is suitably within the range of, for example, 1 to 500 times, and preferably 2 to 200 times, the molar ratio of the acid to 1 mole of the compound [C-1]. The acid that can be used in this elimination reaction is suitably diluted with an appropriate 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 as needed. 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 of 1 mole of the compound [C-1]. Examples of means for supplying a liquid to a supply channel that can be used in this elimination reaction include pumps that are commonly used in this field for supplying liquids, specifically, for example, syringe pumps, plunger pumps, diaphragm pumps, and gear pumps. Examples of flow reactors that can be used in this 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 that can be used for the supply channel and reaction channel 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 selected appropriately, for example, from a range of 0.1 mm to 1.0 mm, and is preferably selected 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], by using a reaction promoter, compound [A-1] and compound [B-1] are subjected to a condensation reaction to produce compound [C-1], and a solution containing an acid is added to the reaction mixture, thereby obtaining Q 1 In the method for producing compound [C-1], the Q of compound [A-1-1] can be obtained by using a reaction promoter. 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 commonly used in the relevant technical field, and a single solvent may be used, or two or more solvents may be mixed. Examples 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. Two or more of these solvents may be mixed and used. Solvents that can be used in this continuous reaction include, for example, polar solvents and halogenated solvents. The solvent that can be used in this continuous reaction is not particularly limited, but for example, a mixed solvent of a polar solvent and a halogenated solvent can be used. The lower limit of the proportion of the polar solvent in the mixed solvent of a polar solvent and a halogenated solvent is 1.0 wt%, preferably 2.0 wt%, more preferably 3.0 wt%, and even more preferably 5.0 wt%. The upper limit is 90 wt%, preferably 75 wt%, more preferably 50 wt%, even more preferably 40 wt%, and particularly preferably 30 wt%. Furthermore, these upper and lower limits can be used in appropriate combinations. The proportion of the polar solvent in the mixed solvent of a polar solvent and a halogenated solvent is, for example, suitably in the range of 1% to 50%, preferably 1% to 40%, and particularly preferably 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 that can be removed under acidic conditions; W is an oxygen atom or a sulfur atom; X is di(C alkyl)amino or a group represented by the general formulas [2-1] to [2-8]: [ka] [wherein * represents the bonding position with P] Preferably, it is di(C alkyl)amino, more preferably dimethylamino; G is a group of the general formula [7]: [ka] (In the formula, * indicates the binding position with T; Z is a group of the general formula [8A] to [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 are represented by the following general formula [9A]: [ka] (In the formula, * represents a bond position; and R 9 represents a long chain alkyl and / or a long chain alkenyl. represents a substituent represented by the formula: R 8d are the same or different and represent a hydrogen atom, a halogen atom, a long-chain alkyl optionally substituted with 1 to 13 halogen atoms, or a long-chain alkyloxy optionally substituted with 1 to 13 halogen atoms; R8e teeth, (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; and R 8f teeth, (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) represents a long-chain alkenyl-carbonyl. is a substituent represented by the formula: L is the general formula

[10] : [ka] (In the formula, * indicates the bonding position with Z; ** indicates the bond position with the oxygen atom; and L 1 is optionally substituted C 2-10 Alkylene or optionally substituted C 6-10 represents arylene.) is a substituent represented by the 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 with 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 is eliminated, Formula [A-1]: [ka] [In the formula, B P, W, X, G, T, and n are as defined above.] The compound 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 between 1 and 10. and reacting 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 also includes a method for preparing a compound of formula (I).

[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, This can include removing Q1. The continuous reaction can be carried out in a flow reactor. 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 eliminate Q1 to form a compound of the formula [A-1], and A method of preparing the compound of general formula [C-1] by supplying a solution containing the compound of general formula [A-1] and a solution containing the compound of general formula [B-1] to a subsequent flow reactor is exemplified. In some cases, a flow reactor can 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 Desorption method Since compound [C-2] is an unstable compound, Q substituted at the 5'-position oxygen atom of the 5'-terminal nucleoside of compound [C-2] is 1 Before the elimination of the aryl group, 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 the 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 oxidizing agents 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; and 5-phenyl-3H-1,2,4-dithiazol-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 dissolved in an appropriate solvent.

[0077] In the compound [C-2], when G is (1) a silicon substituent, (2) a long-chain alkylcarbonyl, (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 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 oxidizing agents 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, Q substituted on the oxygen atom at the 5'-position of the nucleoside unit on the 5'-terminal side of compound [D-2] is obtained. 1 From compound [D-2], Q in the molecule can be eliminated. 1 By eliminating the group, 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 support, the purification can be carried out, for example, by filling compound [D-2] into an appropriate column and eluting it 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), the reaction can be carried out, for example, 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 supply channels into a flow reactor or reaction channel, 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 compound [C-1], compound [D-2], compound [E-1], or compound [E-2] contains protecting groups in its molecule, a deprotection treatment appropriate for the type or nature of the protecting groups can be performed to produce a compound from which all protecting groups have been removed. For example, all protecting groups in the compound can be removed according to the deprotection method described in "Green's Protective Groups in Organic Synthesis, 4th Edition, 2006." Specifically, the protecting groups for the substituent [6] and the amino or hydroxyl groups of the nucleic acid bases in compound [C-1], compound [D-2], compound [E-1], or compound [E-2] can be removed by treating the compound with (1) aqueous ammonia, (2) aqueous ammonia / ethanol, or (3) a mixed solution of aqueous ammonia and methylamine. In addition, for example, the Q 1 The same as the "acid" described in the "Method for removing Q in the molecule of compound [C-2]" 1 The groups that can be removed under acidic conditions, which are substituted on the protecting group of the amino group at the 3'-position of the nucleoside at the 3'-terminus of compound [C-1] and the hydroxyl group at the 5'-position of the nucleoside at the 5'-terminus of compound [D-2], can be removed by treating with the same "acid" described in "Step 2: Production of compound [E-2]" of "Method for removing compound [C-1]" or a solution of hydrochloric acid or acetic acid diluted with an appropriate solvent. After removing the protecting group of the nucleic acid base portion, if the group that can be removed under acidic conditions and is substituted on the 5'-position hydroxyl group of the 5'-terminal nucleoside of compound [D-2] is to be removed, an acid diluted with water is used, and if the nucleic acid base portion is protected, an acid diluted with an appropriate organic solvent is used.

[0082] (I) Purification and separation process The compound [C-1] or compound [E-1] from which all the protecting groups have been removed can be separated from the reaction mixture by a conventional separation and purification method, for example, extraction, concentration, neutralization, filtration, centrifugation, recrystallization, or separation of C8 to C 18 The protein can be isolated by using, alone or in combination, means such as reverse phase column chromatography, cation exchange column chromatography, anion exchange column chromatography, gel filtration column chromatography, high performance liquid chromatography, dialysis, and ultrafiltration (see, for example, International Publication No. 1991 / 09033A1). When the desired compound is purified using reverse phase chromatography, a mixed solution of 20 mM triethylamine / acetic acid buffer and acetonitrile can be used as the elution solvent, for example. Furthermore, when purifying the desired compound using ion exchange chromatography, 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] The compound [D-2] or compound [E-2] from which all the protecting groups have been removed can be separated from the reaction mixture by conventional separation and purification means, for example, extraction, concentration, neutralization, filtration, centrifugation, recrystallization, C8 to C 18 Reversed-phase column chromatography, C8 to C 18 Isolation and purification can be achieved by using 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, either alone or in combination. Examples of the "elution solvent" include acetonitrile, methanol, ethanol, isopropyl alcohol, or water, or a mixture of these solvents in any ratio. In this case, additives such as sodium phosphate, potassium phosphate, sodium chloride, potassium chloride, ammonium acetate, triethylammonium acetate, sodium acetate, potassium acetate, Tris-HCl, or ethylenediaminetetraacetic acid can be added at a concentration of 1 mM to 2 M to adjust the pH of the solution 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 the hydroxyl group of a compound corresponding to compound [A] according to a known method.

[0085] Representative examples are introduced below to explain the method for producing 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, by 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 represents 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 5'-terminal hydroxyl group of a compound represented by the above general formula

[21] (hereinafter referred to as "compound

[21] ") using a compound represented by the above general formula [20A] (hereinafter referred to as "compound [20A]"). This reaction for introducing a silicon substituent can be carried out in accordance with 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 same as that described above in "Q in the molecule of compound [C-1]." 1 Examples of the "acid" include the same as those described in "Method for removing the acid". The amount of acid that can be used in this step is, for example, in a molar ratio of 1 to 500 times, and preferably 2 to 200 times, per mole of compound [A-1a-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, and examples thereof include, but are not limited to, 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 as needed. The "scavenger" that can be used in this step is the "Q in the molecule of compound [C-1]" 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, for example, in a molar ratio of 1 to 100 times, preferably 1 to 50 times, per mole of compound [A-1a-Q1].

[0091] (ii) Preparation of compound [A-1], wherein 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 compound [A-1] 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. An example of a method for producing the compound [A-1b] will be 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 [A-1b-Q1] can be produced by condensing compound

[21] with a compound represented by the above general formula [20B] (hereinafter referred to as "compound [20B]"). 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 using a condensing agent in the presence or absence of a base at a temperature within the range of -20°C to 100°C. When a compound [20B] in which Y is a halogen atom 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. Condensing agents that can be used in this step include, for example, 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, and diethyl ether; amides such as dimethylformamide and dimethylacetamide; nitriles such as acetonitrile and propionitrile; hydrocarbons such as benzene and toluene; halogenated hydrocarbons such as chloroform and methylene chloride; and mixed solvents thereof. When a compound [20B] in which Y is a hydroxyl group is used in this step, an additive can be used as needed. 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 condensing agent used are, for example, suitably within a range of 1 to 1.5 moles per mole of compound [20B]. The amount of the base used is, for example, within a range of 1 to 10 equivalents, preferably within a 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 same as that described above in "Q in the molecule of compound [C-1]." 1 Examples of the "acid" include the same as those described in "Method for removing the acid". The amount of acid that can be used in this step is suitably within the range of, for example, 1 to 500 times, and preferably 2 to 200 times, the molar ratio of the compound [A-1b-Q1] to 1 mole. The acid that can be used in this step may be diluted with an appropriate solvent, and examples thereof include, but are not limited to, 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 as needed. The "scavenger" that can be used in this step is the "Q in the molecule of compound [C-1]" 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, for example, in a molar ratio of 1 to 100 times, preferably 1 to 50 times, per mole of compound [A-1b-Q1].

[0097] (iii) Preparation of compound [A-1], wherein 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 compound [A-1] 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] . An example of a method for producing the 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] "). Although compound [20C] is a carboxylic acid, its reactive derivative can also be used in this step. Examples of reactive derivatives of compound [20C] include those commonly used in ester condensation reactions, such as acid halides (e.g., acid chlorides and acid bromides). Compound

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

[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 [A-1c-Q1] can be produced by condensing compound

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

[25] (hereinafter referred to as "compound

[25] "). The condensation reaction and deprotection reaction can be carried out according to known methods.

[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 same as that described above in "Q in the molecule of compound [C-1]." 1 Examples of the "acid" include the same as those described in "Method for removing the acid". The amount of acid that can be used in this step is, for example, in a molar ratio of 1 to 500 times, and preferably 2 to 200 times, per mole of compound [A-1c-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, and examples thereof include, but are not limited to, 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 as needed. The "scavenger" that can be used in this step is the "Q in the molecule of compound [C-1]" 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, for example, in a molar ratio of 1 to 100 times, preferably 1 to 50 times, per mole of 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 the 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]") Compound [A-1d-Q1] can be produced by condensing a compound represented by the above general formula [20D] (hereinafter referred to as "compound [20D]") with a compound represented by the above general formula

[21] (hereinafter referred to as compound

[21] ). The condensation reaction can be carried out in accordance with a method known per se. Although compound [20D] is a carboxylic acid, its reactive derivative can also be used in this step. Examples of reactive derivatives of compound [20D] include those commonly used in ester condensation reactions, such as acid halides (e.g., acid chlorides and 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. Condensing agents that can be used in this step include, for example, 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, and diethyl ether, amides such as dimethylformamide and dimethylacetamide, nitriles such as acetonitrile and propionitrile, hydrocarbons such as benzene and toluene, halogenated hydrocarbons such as chloroform and methylene chloride, and mixed solvents thereof. Furthermore, additives 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 condensing agent used are, for example, suitably within a range of 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 same as that described above in "Q in the molecule of compound [C-1]." 1 Examples of the "acid" include the same as those described in "Method for removing the acid". The amount of acid that can be used in this step is, for example, in a molar ratio of 1 to 500 times, and preferably 2 to 200 times, the amount of compound [A-1d-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, and examples thereof include, but are not limited to, 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 as needed. The "scavenger" that can be used in this step is the "Q in the molecule of compound [C-1]" 1 Examples of the "scavenger" include the same ones as those described in "Method for removing the scavenger". The amount of scavenger that can be used in this step is, for example, in a molar ratio of 1 to 100 times, preferably 1 to 50 times, the amount of scavenger per mole of 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 a compound represented by the above general formula

[26] (hereinafter referred to as "compound

[26] ") with a 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 are 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] Furthermore, 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 aminating 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 alkyl, and 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 in accordance with 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, then 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 known methods. (e) For example, by alkylating one of the hydroxyl groups of ethane-1,2-diol with an alkyl halide, 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 is carbonylated with a long chain alkyl, where Z is a substituent [8E] and R 8e is a long-chain alkyl-carbonyl, a compound

[26] can be produced. The compound used for the long-chain alkyl-carbonylation can be, for example, the corresponding carboxylic acid compound or a reactive derivative thereof. Examples of reactive derivatives include those typically used in ester condensation reactions, such as acid halides (e.g., acid chlorides and acid bromides). For example, by condensing one hydroxyl group of ethane-1,2-diol with compound [20C], 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 by the same method as that for preparing compound (18) in which Z is a substituent [8E], except that 2-amino-ethanol is used 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], followed by removing the 9-fluorenylmethyloxycarbonyl group with piperidine. The condensation reaction and the deprotection reaction of the 9-fluorenylmethyloxycarbonyl group can be carried out according to known methods. (h) For example, compound

[26] in which Z is a substituent [8H] can be produced by using 1-(tert-butoxycarbonyl)piperazine instead of ethane-1,2-diol in the same manner as in the production method of compound

[26] in which Z is a substituent [8E], 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), WO 2014 / 189142A1, WO 2016 / 060135A1, WO 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, by treating 9H-xanthen-9-one having a hydroxyl group with a base such as sodium hydride and then reacting with an appropriate long-chain alkyl halide, it is possible to prepare the corresponding 9H-xanthen-9-one having a long-chain alkyloxy group. Furthermore, by reacting with optionally substituted phenylmagnesium bromide, it is possible to prepare compound

[26] in which Z is a substituent [8M]. Furthermore, by adjusting 9H-xanthen-9-one derivatives or phenylmagnesium bromide derivatives having various substituents according to known methods, it is possible to prepare the desired compound

[26] in which Z is a substituent [8M].

[0112] Furthermore, compound

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

[30] can be produced by acetylating the compound represented by the above general formula

[29] (see, for example, WO 91 / 09033A1) with acetic anhydride in the presence of a base. 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 acid. 1 The elimination reaction of the formula (1) 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, WO 91 / 09033 A1). Compound

[32] can be produced, for example, by a known method (see, for example, WO 91 / 09033 A1).

[0116] Step 4: Preparation of compound

[21] For example, compound

[21] can be produced by selectively removing the acetyl group from compound

[33] using an alkali metal alkoxide such as sodium methoxide. The acetyl group removal reaction can be carried out according to a known method (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] in which 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 using compound [20A] in a compound represented by the above general formula

[34] (hereinafter referred to as "compound

[34] ") to introduce a silicon substituent into the 3'-hydroxyl group of the 3'-terminal nucleoside unit of compound

[34] . This reaction for introducing a silicon substituent can be carried out in accordance with 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 same as the "Q in the molecule of compound [D-2]" 1 Examples of the "acid" include the same as those described in the "Elimination reaction of the hydroxyl group with hydroxyl group". The amount of acid that can be used in this step is, for example, in a molar ratio of 1 to 500 times, and preferably 2 to 200 times, per mole of compound [A-2a-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, and examples thereof include, but are not limited to, 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 as needed. The "scavenger" that can be used in this step is the "Q in the molecule of compound [D-2]" 1 Examples of the "scavenger" include the same ones as those described in the "elimination reaction of the hydroxyl group". The amount of the scavenger that can be used in this step is, for example, in a molar ratio of 1 to 100 times, preferably 1 to 50 times, per mole of 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 [A-2b-Q1] can be produced by condensing compound [20B] with compound

[34] . The condensation reaction can be carried out in accordance with a known method. When a compound [20B] in which Y is a hydroxyl group is used in this step, the reaction can be carried out using a condensing agent in the presence or absence of a base 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. Condensing agents that can be used in this step include, for example, 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, and diethyl ether; amides such as dimethylformamide and dimethylacetamide; nitriles such as acetonitrile and propionitrile; hydrocarbons such as benzene and toluene; halogenated hydrocarbons such as chloroform and methylene chloride; and mixed solvents thereof. When a compound [20B] in which Y is a hydroxyl group is used in this step, an additive can be used as needed. 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 the compound [20B] and the condensing agent used are, for example, suitably within a range of 1 to 1.5 moles per mole of the compound

[34] . The amount of the base used is, for example, within the range of 1 to 10 equivalents, preferably within the range of 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 same as the "Q in the molecule of compound [D-2]" 1 Examples of the "acid" include the same as those described in the "Elimination reaction of the hydroxyl group with hydroxyl group". The amount of acid that can be used in this step is, for example, in a molar ratio of 1 to 500 times, and preferably 2 to 200 times, the amount of compound [A-2b-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, and examples thereof include, but are not limited to, 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 as needed. The "scavenger" that can be used in this step is the "Q in the molecule of compound [D-2]" 1 Examples of the "scavenger" include the same ones as those described in the "elimination reaction of the hydroxyl group". The amount of the scavenger that can be used in this step is, for example, in a molar ratio of 1 to 100 times, preferably 1 to 50 times, per mole of compound [A-2b-Q1].

[0129] (iii) Preparation of compound [A-2], wherein 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 known methods.

[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 known method. Examples of the "oxidizing agent" include iodine and tert-butyl hydroperoxide. The oxidizing agent usable in this step can also be diluted with an appropriate solvent to a concentration of 0.05 to 2 M. The solvent is not particularly limited, but examples include pyridine, tetrahydrofuran, water, and a mixture thereof. For example, iodine / water / pyridine-tetrahydrofuran or iodine / pyridine-acetic acid, or a peroxidizing agent (tert-butyl hydroperoxide / methylene chloride, etc.) 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 minute 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 same as the "Q in the molecule of compound [D-2]" 1 Examples of the "acid" include the same as those described in the "Elimination reaction of the hydroxyl group with hydroxyl group". The amount of acid that can be used in this step is, for example, in a molar ratio of 1 to 500 times, and preferably 2 to 200 times, per mole of compound [A-2c-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, and examples thereof include, but are not limited to, 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 as needed. The "scavenger" that can be used in this step is the "Q in the molecule of compound [D-2]" 1 Examples of the "scavenger" include the same ones as those described in the "elimination reaction of the hydroxyl group". The amount of the scavenger that can be used in this step is, for example, in a molar ratio of 1 to 100 times, preferably 1 to 50 times, per mole of compound [A-2c-Q1].

[0135] (iv) Preparation of compound [A-2], wherein 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 [20D] can be condensed with the compound

[34] to produce the compound represented by the general formula [A-2d-Q1] (hereinafter referred to as "compound [A-2d-Q1]"). The condensation reaction can be carried out according to a method known per se. Although compound [20D] is a carboxylic acid, its reactive derivative can also be used in this step. Examples of reactive derivatives of compound [20D] include those commonly used in ester condensation reactions, such as acid halides (e.g., acid chlorides and 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. Condensing agents that can be used in this step include, for example, 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, and diethyl ether, amides such as dimethylformamide and dimethylacetamide, nitriles such as acetonitrile and propionitrile, hydrocarbons such as benzene and toluene, halogenated hydrocarbons such as chloroform and methylene chloride, and mixed solvents thereof. Furthermore, additives 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 the compound [20D] and the condensing agent used are, for example, suitably within a range of 1 to 1.5 moles per mole of the compound

[34] . The amount of the base used is, for example, within the range of 1 to 10 equivalents, preferably within the range of 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 same as the "Q in the molecule of compound [D-2]" 1 Examples of the "acid" include the same as those described in the "Elimination reaction of the hydroxyl group with hydroxyl group". The amount of acid that can be used in this step is, for example, in a molar ratio of 1 to 500 times, and preferably 2 to 200 times, the amount of compound [A-2d-Q1]. The acid that can be used in this step may be diluted with an appropriate solvent, and examples thereof include, but are not limited to, 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 as needed. The "scavenger" that can be used in this step is the "Q in the molecule of compound [D-2]" 1 Examples of the "scavenger" include the same ones as those described in the "elimination reaction of the hydroxyl group". The amount of scavenger that can be used in this step is, for example, in a molar ratio of 1 to 100 times, preferably 1 to 50 times, the amount of scavenger per mole of 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 the compound is nucleoside unit [4a], but compounds in which all or part of nucleoside unit [4a] is replaced with nucleoside unit [4b] or nucleoside unit [4c] can also be produced using the same method as above.

[0141] (K) Preparation 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] according to a known method.

[0142] Representative examples are introduced below to explain the method for producing 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 following method. [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 a 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 known method (see, for example, U.S. Patent Application Publication No. 2014 / 0330006A1, WO 2012 / 043730A1, WO 2013 / 082548A1, and WO 91 / 09033A1). The compound

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

[21] .

[0146] (K-2) Preparation 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] "). This reaction can be carried out according to a method known per se (see, for example, Helvetica Chimica Acta, Vol. 70, pp. 175-186 (1987), International Publication No. 2003 / 106468A1, Acta Nature, 6, pp. 116-118 (2014), and Russian Journal of General Chemistry, Vol. 67, No. 1, pp. 62-64 (1997)).

[0148] Step 2: Preparation of Compound [B-2] According to a known method, compound

[42] is reacted 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'-hydroxyl group of the 3'-terminal nucleoside unit, thereby producing compound "B-2". 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 compound

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

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

[0149] Compound [B-2] is a compound in which each nucleoside unit constituting the compound is a nucleoside unit [4e]. However, a compound in which all or part of the nucleoside unit [4e] is replaced with a nucleoside unit [4f] or a 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. The "conversion yield (%)" means the ratio of the raw material converted into the target product, and is calculated by "{peak area (%) corresponding to the target product detected by high performance liquid chromatography (hereinafter referred to as "HPLC")} ÷ {peak area (%) corresponding to the raw material detected by HPLC + peak area (%) corresponding to the target product detected by HPLC} × 100". HPLC conditions: 0.5 mg of the product was dissolved in acetonitrile or an 80% aqueous acetonitrile solution, and HPLC analysis was 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 (0-20 min), 95%B (20-25 min), 70%B (25-35 min) 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 aqueous ammonia Mobile phase: MeOH Gradient: 70-95%B (8 min) Detector 1: UV 264 nm Detector 2: Quadrupole time-of-flight mass spectrometer Ionization method: ESI+ Measurement range: 100-2000 m / z

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

[0152] Step 2: Preparation of [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl 4-(octadecylamino)-4-oxobutanoate (hereinafter referred to as "G1-suc-morT-OFF") To a solution of G1-suc (14.4 g) in tetrahydrofuran (150 mL), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (8.56 g, 1.2 eq.) was added and stirred at room temperature. Subsequently, 1-((2R,6S)-6-(hydroxymethyl)-4-tritylmorpholin-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (hereinafter referred to as "morT-OH") (18 g, 1.0 eq.) and 4.57 g of 4-(N,N-dimethylamino)pyridine were added and stirred in a water bath at 70°C for 1 hour. After cooling to room temperature, 0.1 M aqueous sodium dihydrogen phosphate was added and stirred for a while. The aqueous layer was removed, and the organic layer was washed once with 0.1 M aqueous sodium dihydrogen phosphate and once with saturated brine diluted twice with water. The combined aqueous layers were extracted with dichloromethane, and the combined organic layers were dried over anhydrous sodium sulfate. After filtration, the solvent was evaporated and the residue was dried under reduced pressure to give 4-(octadecylamino)-4-oxobutanoate [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methyl (hereinafter referred to as "G1-suc-morT-ON") (white amorphous, 28.1 g, 89.5%). This was dissolved in 140 mL of dichloromethane and stirred in an ice bath. 20 mL of 2,2,2-trifluoroethanol and 10.3 mL of triisopropylsilane were added. After stirring for a while, 5.1 mL of trifluoroacetic acid was added dropwise. One hour after the dropwise addition, the reaction mixture was poured into 100 mL of saturated aqueous sodium bicarbonate solution with ice while stirring. After confirming that the aqueous layer had a pH of 7-8, the aqueous layer was extracted with dichloromethane. The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was performed by column chromatography on silica gel using a dichloromethane-methanol mixture as the mobile phase, and the residue was dried under reduced pressure to obtain 19.89 g of G1-suc-morT-OFF as a powder. 1H-NMR(CDCl3):δ8.90(1H,bs);7.25(1H,d,J=1.6Hz);5.72(1H,dd,J=9.6Hz,2.6Hz);5 .65(1H,m);4.14(2H,d,J=5.2Hz);3.98(1H,m);3.23(2H,dd,J=12.8Hz,7.0Hz);3.12(2 H,dd,J=12Hz,2.6Hz);2.95(2H,dd,J=12.8Hz,1.8Hz);2.60~2.75(4H,m);2.47(2H,t,J =6.8Hz);1.95(3H,d,J=1.6Hz);1.48(2H,m), 1.21~1.34(29H,m);0.88(3H,t,J=6.4Hz) ESI-MS(+):593.36(M+H)

[0153] Example 2 {[(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1-yl)morpholin-2-yl]methyl}{2-octadecanoyloxy-1-[(octadecanoyloxymethyl)ethyl]} succinate (hereinafter referred to as "G2-suc-morT-OFF") Step 1: Preparation of 4-((1,3-bis(stearoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (hereinafter referred to as "G2-suc") Dichloromethane (8 mL) was added to 1 g (1.60 mmol) of 2-hydroxypropane-1,3-diyl distearate, followed by 176 mg (1.76 mmol) of succinic anhydride and 293 mg (2.40 mmol) of 4-(N,N-dimethylamino)pyridine, and the mixture was stirred at room temperature for 16 hours. After the reaction was completed, 1 M aqueous sodium dihydrogen phosphate solution was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated to give G2-sucrose (1.40 g).

[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 (5.2 mL) was added to G2-suc (900 mg, 1.24 mmol) and 277 mg (1.45 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, followed by 500 mg (1.03 mmol) of morT-OH and 132 mg (1.09 mmol) of 4-(N,N-dimethylamino)pyridine. The mixture was stirred at room temperature for 16 hours. After the reaction was complete, 0.1 M aqueous sodium dihydrogen phosphate was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give G2-suc-morT-ON (1.09 g, 89%). 1H-NMR(CDCl3):δ8.04(1H,s);7.17~7.51(15H,m);6.98(1H,s);6.12(1H,dd,J=9.6H z,2.4Hz);5.25(1H,m);4.34~4.37(1H,m);4.26~4.30(2H,m);4.11~4.16(2H,m);4. 00~4.08(2H,m);3.35(1H,d,J=11.2Hz);3.10(1H,d,J=11.6Hz);2.60(4H,s);2.30( 4H,t,J=7.6Hz);1.83(3H,s);1.38~1.44(2H,m);1.24(60H,m);0.87(6H,t,J=6.8Hz)

[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. Next, 127 μL (0.62 mmol) of triisopropylsilane and 64 μL (0.82 mmol) of trifluoroacetic acid were added at 0°C and stirred at room temperature for 1 hour. After the reaction was completed, saturated aqueous sodium bicarbonate solution was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give G2-suc-morT-OFF (373 mg, 95%). 1 H-NMR(CDCl3):δ8.04(1H,bs);7.24(1H,s);5.70(1H,d,J=2Hz);5.21~5.26(1H,m);4.28~4.31(2H,m);4.13~4.17(4H,m);3.96~4.00(1H,m);3.1 1(1H,dd,J=12.4,2Hz);2.94(1H,dd,J=12.8,2.4Hz);2.57~2.65(6H,m);2.32(4H,t,J=7.6Hz);1.95(3H,s);1.25(60H,m);0.88(6H,t,J=7.6Hz)

[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]succinate (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") 4-((1,3-bis(oleoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (hereinafter referred to as "G3-suc") was produced using 2-hydroxypropane-1,3-diyl dioleate as a raw material in the same manner as in Step 1 of Example 2. G3-suc-morT-ON was then produced in the same manner as in Step 2 of Example 2. 1 H-NMR(CDCl3):δ8.00(1H,s);7.17~7.51(15H,m);6.99(1H,s);6.09~6.12(1H,m);5.29~5.3 8(4H,m);5.20~5.25(1H,m);4.33~4.37(1H,m);4.26~4.30(2H,m);4.12~4.16(2H,m);4.00~ 4.09(2H,m);3.35(1H,d,J=11.6Hz);2.15(1H,d,J=11.6Hz);2.60(4H,m);2.30(4H,t,J=7.2 Hz);1.97~2.02(8H,m);1.83(3H,s);1.57~1.61(2H,m);1.28(44H,m);0.89(6H,t,J=6.8Hz)

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

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

[0159] Step 2: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl 4-oxo-4-(4-stearoylpiperazin-1-yl)butanoate (hereinafter referred to as "G4-suc-morT-ON") Tetrahydrofuran (10 mL) was added to G4-suc (982 mg, 2.17 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (555 mg, 2.90 mmol) and stirred at 70 °C. Next, morT-OH (1 g, 2.07 mmol) and 4-(N,N-dimethylamino)pyridine (265 mg, 2.17 mmol) were added and stirred at 70 °C for 30 minutes. After the reaction was complete, the mixture was allowed to cool to room temperature. 0.1 M aqueous sodium dihydrogen phosphate solution was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give G4-suc-morT-ON (1.68 g, 89%). 1 H-NMR(CDCl3):δ8.00(1H,s);7.16~7.50(15H,m);6.97(1H,s);6.10(1H,d ,J=8Hz);4.34~4.36(1H,m);4.04(2H,d,J=4.8Hz);3.57~3.64(4H,m);3.4 4~3.48(4H,m);3.32~3.34(1H,m);3.09~3.12(1H,m);2.60~2.64(4H,m);2 .31(2H,t,J=7.6Hz);1.82(3H,s);1.23~1.42(32H,m);0.86(3H,t,J=6.8)

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

[0161] Example 5 [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl 4-(octadecylcarbamoyl)benzoate (hereinafter referred to as "G5-tpa-morT-OFF") Step 1: Preparation of [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methyl 4-(octadecylcarbamoyl)benzoate (hereinafter referred to as "G5-tpa-morT-ON") This was prepared in the same manner as in Step 2 of Example 2 using 4-(octadecylcarbamoyl)benzoic acid. 1 H-NMR(CDCl3):δ8.24(1H,s);7.97(2H,d,J=8Hz);7.77(2H,d,J=8Hz);7.17~7.46(15H,m);6.95(1H,s);6.12~6.16(1H,m);4.49~4.51(1 H,m);4.25~4.33(2H,m);3.42~3.47(2H,m);3.35~3.38(1H,m);3.21~3.24(1H,m);1.79(3H,s);1.23~1.44(34H,m);0.86(3H,t,J=6.8Hz)

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

[0163] Example 6 {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl 4-(4-(4-(octadecylcarbamoyl)benzoyl)piperazin-1-yl)-4-oxobutanoate (hereinafter referred to as "G6-suc-morT-OFF") Step 1: Preparation of 4-(4-(4-(octadecylcarbamoyl)benzoyl)piperazin-1-yl)-4-oxobutanoic acid (hereinafter referred to as "G6-suc") This was produced in the same manner as in step 1 of 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. 1H-NMR(CDCl3):δ8.08(1H,bs);7.81(2H,d,J=7.6Hz);7.16~7.50(17H,m);6.97(1H,s);6.08~6.10(1H,m);4.33~4.39(1H,m);4.0 2~4.04(2H,m);3.31~3.79(11H,m);3.08~3.11(1H,m);2.60~2.69(4H,m);1.81(3H,s);1.23~1.44(34H,m);0.86(3H,t,J=6.4Hz)

[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. Next, 145 μL (0.70 mmol) of triisopropylsilane and 53 μL (0.70 mmol) of trifluoroacetic acid were added at 0°C and stirred at room temperature for 1 hour. After the reaction was completed, saturated aqueous sodium bicarbonate solution was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give G6-suc-morT-OFF (372 mg; 98%). 1 H-NMR(CDCl3):δ8.05(1H,bs);7.79(2H,d,J=7.6Hz);7.45(2H,d,J=7.6Hz );7.23(1H,s);6.08~6.11(1H,m);5.67~5.69(1H,m);4.10~4.15(2H,m);3 .96~3.99(1H,m);3.36~3.79(8H,m);3.08~3.11(1H,m);2.93~2.96(1H,m) ;2.57~2.70(6H,m);1.92(3H,s);1.23~1.38(34H,m);0.86(3H,t,J=7.2Hz)

[0166] Example 7 {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl 3,4,5-tris(octadecyloxy)benzoate (hereinafter referred to as "G7-morT-OFF") Step 1: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-trityl-morpholin-2-yl}methyl 3,4,5-tris(octadecyloxy)benzoate (hereinafter referred to as "G7-morT-ON") This was prepared in the same manner as in Step 2 of Example 2 using 3,4,5-trioctadecoxybenzoic acid. 1H-NMR(CDCl3):δ7.90(1H,bs);7.12~7.45(17H,m);6.97(1H,s);6.12~6.14(1H,m);4.46~4.51(1H,m);4.28~4.32(1H,m);4.16~4 .20(1H,m);3.90~4.00(6H,m);3.37~3.40(1H,m);3.22~3.25(1H,m);1.78~1.82(5H,m);1.23~1.50(96H,m);0.86(9H,t,J=6.8Hz)

[0167] Step 2: Production of G7-morT-OFF It was produced in the same manner as in step 3 of Example 2. 1 H-NMR(CDCl3): δ7.98(1H,bs);7.22(3H,m);5.69~5.72(1H,m);4.32~4.36(2H,m);4.08~4.12(1H,m);3.94~4.01(6H ,m);3.11~3.14(1H,m);3.02~3.05(1H,m);2.64~2.72(2H,m);1.90(3H,m);1.23~1.45(96H,m)0.86(9H,t,J=7.2Hz)

[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 1.5 g (1.60 mmol) of 3,4,5-trioctadecyloxybenzoic acid, 370 mg (1.90 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, and 240 mg (1.90 mmol) of 4-(N,N-dimethylamino)pyridine were added to 8.1 mL of chloroform, followed by 120 mg (1.90 mmol) of ethylene glycol, and the mixture was stirred at room temperature for 3 hours. After the reaction was completed, 1 M aqueous sodium dihydrogen phosphate was added to the reaction mixture, which was then extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give 2-hydroxyethyl 3,4,5-trioctadecyloxybenzoate (882 mg; 56%). 1H-NMR(CDCl3):δ7.26(2H,s);4.45~4.47(2H,m);3.95~4.03(8H,m);1.25~1.52(96H,m);0.88(9H,t,J=7.2Hz)

[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 the same manner as in Step 1 of Example 2, and then G8-suc-morT-ON was obtained in the same manner as in Step 2 of Example 2. 1 H-NMR(CDCl3):δ7.87(1H,bs);7.12~7.43(17H,m);6.97(1H,s);6.07~6.10(1H,m);4.33~4.46(5H,m);3.91~4.07(8H,m );3.31~3.34(1H,m);3.07~3.10(1H,m);2.56~2.60(4H,m);1.68~1.80(5H,m);1.23~1.50(96H,m);0.86(9H,t,J=7.2Hz)

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

[0171] Example 9 {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl 4-(dioctadecylamino)-4-oxobutanoate (hereinafter referred to as "G9-suc-morT-OFF") Step 1: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl 4-(dioctadecylamino)-4-oxobutanoate (hereinafter referred to as "G9-suc-morT-ON") Using N-octadecyloctadecane-1-amine as a starting material, 4-(dioctadecylamino)-4-oxobutanoic acid (hereinafter referred to as "G9-suc") was produced in the same manner as in Step 1 of Example 2. G9-suc-morT-ON was then produced in the same manner as in Step 2 of Example 2. 1 H-NMR(CDCl3):δ7.88(1H,bs);7.17~7.43(15H,m);6.98(1H,s);6.06~6.09(1H,m);4.31~4.35(1H,m);4.01 ~4.03(2H,m);3.08~3.34(8H,m);2.52~2.64(4H,m);1.82(3H,s);1.23~1.52(64H,m);0.85(6H,t,J=6.8Hz)

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

[0173] Example 10 {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl 4-[{1-(octadecylamino)-1-oxo-3-phenylpropan-2-yl}amino]-4-oxobutanoate (hereinafter referred to as "G10-suc-morT-OFF") Step 1: Preparation of {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl 4-[{1-(octadecylamino)-1-oxo-3-phenylpropan-2-yl}amino]-4-oxobutanoate (hereinafter referred to as "G10-suc-morT-ON") To 500 mg (1.88 mmol) of 2-tert-butoxycarbonylamino-3-phenyl-propanoic acid, 9.4 mL of tetrahydrofuran was added, followed by 652 μL (3.77 mmol) of N-ethyl-N-isopropyl-propan-2-amine, 46 mg (0.38 mmol) of 4-(N,N-dimethylamino)pyridine, 505 mg (2.64 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, and octadecane-1-amine. The mixture was stirred at room temperature for 5 hours. After completion of the reaction, 1 M aqueous sodium dihydrogen phosphate solution was added to the reaction mixture, extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The resulting residue was purified by silica gel chromatography to give tert-butoxycarbonylamino-N-octadecyl-3-phenyl-propanamide (779 mg, 80%). 1 H-NMR (CDCl3): δ7.18~7.28(5H,m);5.57(1H,bs);5.06(1H,bs);4.20~4.26(1H ,m);2.95~3.12(4H,m);1.40(9H,s);1.14~1.28(32H,m);0.86(3H,t,J=6.8Hz)

[0174] To 779 mg (1.51 mmol) of tert-butoxycarbonylamino-N-octadecyl-3-phenyl-propanamide, 15 mL of dichloromethane was added, followed by 1.74 mL (22.61 mmol) of trifluoroacetic acid, and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, the solvent was distilled off to obtain 2-amino-N-octadecyl-3-phenyl-propanamide (620 mg). The crude product was subjected to a reaction similar to that in Step 1 of Example 2 to produce G10-suc. The reaction was then performed similar to that in Step 2 of Example 2 to produce G10-suc-morT-ON. 1H-NMR(CDCl3): δ8.08(1H,bs);7.16~7.32(20H,m);6.98(1H,s);6.29~6. 31(1H,m);6.10~6.12(1H,m);5.56~5.59(1H,m);4.51~4.57(1H,m);4.35~ 4.37(1H,m);4.02(2H,d,J=5.6Hz);3.73~3.77(1H,m);2.92~3.33(6H,m); 2.39~2.67(4H,m);1.84(3H,s);1.21~1.45(32H,m);0.88(3H,t,J=7.6Hz)

[0175] Process 2 Manufacturing of G10-suc-morT-OFF The target product was obtained in the same manner as in step 3 of Example 2. 1 H-NMR(CDCl3): δ8.33(1H,bs);7.16~7.32(6H,m);6.38~6.40(1H,m);5.67~5.71(2H,m);4.54~4.58(1H,m);4.08~4.16( 3H,m);3.94~4.01(1H,m);2.91~3.17(5H,m);2.46~2.79(5H,m);1.94(3H,s);1.13~1.36(32H,m);0.88(3H,t,J=7.6Hz)

[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 Preparation of ]-ON (using tetrabutylammonium chloride as a reaction accelerator, reaction time (30 minutes)) [ka] 14.1 mg (0.02 mmol) of G1-suc-morA-OFF, 7.6 μL (0.06 mmol) of N-ethylmorpholine, and 16.7 mg (0.06 mmol) of tetrabutylammonium chloride were dissolved in 200 μL of dichloromethane. 21.7 mg (0.03 mmol) of morA was dissolved in 200 μL of dichloromethane and the mixture was shaken at 40 °C for 30 minutes. After 30 minutes, 160 μL of a 1% 1-methylpiperazine / dichloromethane solution was added to 40 μL of the reaction mixture to terminate the reaction. This solution was further diluted 10-fold with acetonitrile and analyzed by HPLC (raw material Rt: 13.20 min, target product Rt: 19.68 min, 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 replacing morT-OH with N-(9-((2R,6S)-6-(hydroxymethyl-4-tritylmorpholin-2-yl)-9H-purin-6-yl)benzamide (morA-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 a reaction promoter, reaction time was 60 minutes) [ka] G1-suc-morA-OFF (10 mg, 0.0142 mmol), morT (12.9 mg, 0.0212 mmol), N-ethylmorpholine (5.38 μL, 0.0425 mmol), and 1-butyl-1-methylpyrrolidinium chloride (7.55 mg, 0.0425 mmol) were dissolved in dichloromethane (283 μL) and stirred at room temperature for 60 minutes. After 60 minutes, 1-methylpiperazine (4.71 μL, 0.0425 mmol) was added to the reaction mixture to terminate the reaction. This solution was further diluted 60-fold with acetonitrile and analyzed by HPLC (raw material Rt: 13.21 min, target product Rt: 18.77 min, 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 (using 1-butyl-1-methylpyrrolidinium chloride as a reaction promoter) Solution a shown below was pumped at 0.05 mL / min and solution b at 0.2 mL / min. The two solutions were mixed and then reacted in a 30 mL tube reactor at 40°C for 120 minutes. After the reaction, the mixture was mixed with solution c shown below, which was pumped at 0.6 mL / min, and the reaction was carried out in a 5 mL tube reactor at 40°C for 5.9 minutes. The mixture was then mixed with solution d shown below, which was pumped at 0.85 mL / min, and collected in a dropping funnel. The organic layer in the dropping funnel was pumped at 0.8 mL / min and mixed with solution e shown below, which was pumped at 0.4 mL / min, and the reaction was carried out in a 15.6 mL tube reactor at room temperature for 13 minutes. After the reaction, the mixture was further mixed with solution f shown below, which was pumped at a flow rate of 1.5 mL / min, and collected in a separatory funnel. The collected organic layer was washed with solution g and saturated sodium bicarbonate water, dried over sodium sulfate, and the solvent was removed by evaporation. After dissolving it in a small amount of dichloromethane, diisopropyl ether was added, and the resulting precipitate was filtered and dried under reduced pressure overnight to give G1-suc-PMO[A Bz -G CE,Pac -TTTC BzA 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 1 M solution of 1-butyl-1-methylpyrrolidinium chloride in dichloromethane 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. Solution d: Aqueous sodium dihydrogen phosphate solution (1 M) 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. Solution 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 Preparation of ]-ON (using 1-butyl-1-methylpyrrolidinium chloride as a reaction promoter, 1,3-dimethyl-2-imidazolidinone / dichloromethane as a solvent, reaction time (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 1.26 g (0.213 mmol) of [-T]-OFF, 230 mg (0.319 mmol) of morA, 81 μL (0.638 mmol) of N-ethylmorpholine, and 113 mg (0.638 mmol) of 1-butyl-1-methylpyrrolidinium chloride were dissolved in 7.1 mL of 1,3-dimethyl-2-imidazolidinone / dichloromethane solution and stirred at 40 °C for 120 minutes. After 120 minutes, 71 μL (0.638 mmol) of 1-methylpiperazine was added to the reaction mixture to terminate the reaction. This solution was further diluted 60-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 15.16 min, target product Rt: 18.49, 18.72 min, 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 Preparation of -T]-ON (using 1-butyl-1-methylpyrrolidinium chloride as a reaction promoter, 1,3-dimethyl-2-imidazolidinone / dichloromethane as a solvent, reaction time (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 1.17 g (0.209 mmol) of ]-OFF, 191 mg (0.314 mmol) of morT, 79 μL (0.628 mmol) of N-ethylmorpholine, and 186 mg (1.05 mmol) of 1-butyl-1-methylpyrrolidinium chloride were dissolved in 7.0 mL of 1,3-dimethyl-2-imidazolidinone / dichloromethane solution and stirred at 40 °C for 120 minutes. After 120 minutes, 70 μL (0.628 mmol) of 1-methylpiperazine was added to the reaction mixture to terminate the reaction. This solution was further diluted 60-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 15.18 min, target product Rt: 18.17 min, 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 Preparation of ]-ON (using 1-butyl-1-methylpyrrolidinium chloride as a reaction promoter, 1,3-dimethyl-2-imidazolidinone / dichloromethane as a solvent, reaction time (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 1.08 g (0.182 mmol), morA 198 mg (0.274 mmol), N-ethylmorpholine 69 μL (0.547 mmol), and 1-butyl-1-methylpyrrolidinium chloride 162 mg (0.912 mmol) were dissolved in 6.1 mL of 1,3-dimethyl-2-imidazolidinone / dichloromethane solution and stirred at 40 °C for 120 minutes. After 120 minutes, the reaction was stopped by adding 61 μL (0.547 mmol) of 1-methylpiperazine to the reaction mixture. This solution was further diluted 60-fold with 80% acetonitrile and analyzed by HPLC (raw material Rt: 15.02 min, target product Rt: 18.12, 18.32 min, 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 Preparation of ]-ON (using 1-butyl-1-methylpyrrolidinium chloride as a reaction promoter, dichloromethane as a solvent, reaction time (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 2.14 g (0.361 mmol), morA 391 mg (0.542 mmol), N-ethylmorpholine 137 μL (1.08 mmol), and 1-butyl-1-methylpyrrolidinium chloride 193 mg (1.08 mmol) were dissolved in 12 mL of dichloromethane and stirred at 40 °C for 120 minutes. After 120 minutes, the reaction was stopped by adding 120 μL (1.08 mmol) of 1-methylpiperazine to the reaction mixture. This solution was further diluted 60-fold with 80% acetonitrile and analyzed by HPLC (raw material Rt: 15.16 min, target product Rt: 18.49, 18.72 min, 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 a reaction promoter, dichloromethane was used as a 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 2.0 g (0.357 mmol) of ]-OFF, 326 mg (0.536 mmol) of morT, 136 μL (1.07 mmol) of N-ethylmorpholine, and 318 mg (1.79 mmol) of 1-butyl-1-methylpyrrolidinium chloride were dissolved in 11.9 mL of dichloromethane and stirred at 40 °C for 120 minutes. After 120 minutes, 119 μL (1.07 mmol) of 1-methylpiperazine was added to the reaction mixture to terminate the reaction. This solution was further diluted 60-fold with 80% aqueous acetonitrile and analyzed by HPLC (raw material Rt: 15.18 min, target product Rt: 18.17 min, 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 Preparation of ]-ON (using 1-butyl-1-methylpyrrolidinium chloride as a reaction promoter, dichloromethane as a solvent, reaction time (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 1.90 g (0.321 mmol), morA 348 mg (0.481 mmol), N-ethylmorpholine 122 μL (0.962 mmol), and 1-butyl-1-methylpyrrolidinium chloride 285 mg (1.60 mmol) were dissolved in dichloromethane 10.7 mL and stirred at 40 °C for 120 minutes. After 120 minutes, the reaction was stopped by adding 107 μL (0.962 mmol) of 1-methylpiperazine to the reaction mixture. This solution was further diluted 60-fold with 80% acetonitrile and analyzed by HPLC (raw material Rt: 15.02 min, target product Rt: 18.12, 18.32 min, conversion yield 97.0%).

Claims

1. A compound [A] having a hydroxyl group or a primary or secondary amino group and a compound represented by the following general formula [1]: 【Chemical 1】 [In the formula, ** represents the bond position; D is halogen, 5- to 6-membered saturated cyclic amino or di(C １－６ (alkyl)amino; W ０ represents a lone pair of electrons, 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 that can be removed under basic conditions １－６ alkyl)amino, di(amino-C substituted with a group that can be removed under basic conditions １－６ alkyl)amino or the following general formula [2]: 【Chemistry 2】 (In the formula, * indicates the bonding position with P; a represents an integer of 0 to 2; E is CH ２ , CH-A １ or N-A ２ represents; A １ is C １－６ Alkyl, mono-substituted with a group that can be removed 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 which can be removed under basic conditions, mono (C １－６ alkyl)amino, di(C １－６ alkyl)amino, tri(C １－６ alkyl)ammonio, amino substituted with amidino substituted with a group removable under basic conditions, or the following general formula [3]: 【Chemistry 3】 (In the formula, * represents the bonding position with E; b represents an integer of 0 to 2; c represents 0 or 1; R １１ is C １－６ represents alkyl; and M is CH ２ , an oxygen atom, a sulfur atom, or N-(a group that can be removed under basic conditions). represents a substituent represented by the formula: A ２ is C １－６ Alkyl, mono-substituted with a group that can be removed 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 alkyl, a group that can be removed under basic conditions, aryl, or heteroaryl. represents a substituent represented by the following formula: The compound [B] having a substituent containing a phosphorus atom represented by the following general formula [C] is subjected to a condensation reaction. 【Chemistry 4】 [In the formula, W ０ , X has the same meaning as above; A represents a residue obtained by removing one hydrogen atom from a hydroxyl group or a primary or secondary amino group of compound [A]; and B represents a residue obtained by removing the substituent represented by the above general formula [1] from compound [B].] The method for producing compound [C] 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 imidazolium salts, quaternary morpholinium salts, quaternary phosphonium salts, quaternary piperidinium salts, quaternary pyridinium salts, quaternary pyrrolidinium salts, and quaternary sulfonium salts.

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

3. The reaction accelerator is Tetra C １－１８ Alkylammonium chloride, 1-C １－１８ Alkyl-1-C １－１８ Alkylpyrrolidinium chloride, 1-C １－１８ Alkyl-3-C １－１８ The method according to claim 1, wherein the compound is at least one selected from alkylimidazolium chlorides.

4. The reaction accelerator is tetrabutylammonium chloride, tetrabutylammonium bromide, dodecyltrimethylammonium chloride, trioctylmethylammonium chloride, choline 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, 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 accelerator is at least one selected from the group consisting of tetrabutylammonium chloride, 1-butyl-1-methylpyrrolidinium chloride, and 1-methyl-3-N-octylimidazolium chloride.

6. The method according to any one of claims 1 to 5, wherein compound [A] and compound [B] are compounds each containing one to more than one nucleoside unit in the molecule, and when the compound contains more than one nucleoside unit in the molecule, adjacent nucleoside units in the compound are bonded to each other via a phosphorus bond.

7. Nucleoside units constituting the compound [A] may be the same or different and may be represented by the following general formulas [4a] to [4d]: 【Chemistry 5】 [In the formula, *teeth, (1) the bonding position of the phosphorus atom of the phosphorus bond that is bonded to the oxygen atom at the 5' position of the adjacent nucleoside unit; (2) The bonding position with the hydrogen atom or (3) Bonding position to the substituent [6] represents; **teeth, (1) the bonding position of the phosphorus atom of the phosphorus bond that is bonded to the 3'-position oxygen atom or 3'-position nitrogen atom of the adjacent nucleoside unit; (2) The bonding position with the hydrogen atom or (3) Bonding position to the substituent [6] represents; The substituent [6] is represented by the following general formula [6]: 【Chemistry 6】 (In the formula, * represents the bonding position with **O, *O, or *N in the above general formulae [4a] to [4d]; 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]: 【Chemistry 7】 (In the formula, * indicates the bonding 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]: 【Chemistry 8】 (In the formula, * represents the bonding position with L; j represents an integer of 0 to 4; k represents an integer of 0 to 5; R ８ａ is a hydrogen atom or C １－６ represents alkyl; R ８ｂ are the same or different and each represents a long-chain alkyl; R ８ｃ are the same or different and represent the following general formulas [9A] to [9E]: 【Chemistry 9】 (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 atom, a long-chain alkyl optionally substituted with 1 to 13 halogen atoms, or a long-chain alkyloxy optionally substituted with 1 to 13 halogen atoms; R ８ｅ teeth (1) long-chain alkyl, (2) long chain alkyl-carbonyl or (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy; and R ８ｆ teeth, (1) long-chain alkyl, (2) long chain alkyl-carbonyl or (3) represents a long-chain alkenyl-carbonyl. represents a substituent represented by the formula: L represents a group represented by the general formula [10]: 【Chemistry 10】 (In the formula, * represents the bonding position with Z; ** represents the bonding position with T; and L １ is optionally substituted C ２－１０ Alkylene or optionally substituted C ６－１０ represents arylene.) represents a substituent represented by the following formula: represents a substituent represented by the formula: T is a single bond or the following general formula [11]: 【Chemistry 11】 (In the formula, X has the same meaning as above; W is the above W ０ is synonymous with; * represents the bonding position with **O, *O, or *N in the above general formulae [4a] to [4d]; ** represents the bonding position with G; and q represents an integer of 0 to 10. However, when G is a silicon substituent, T is a single bond. represents a substituent represented by the formula: d represents 0 or 1; B Ｐ represents an optionally protected nucleobase; R ４ａ represents 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 and together with the adjacent carbon atom form a carbonyl; and J is an oxygen atom or N-R ４ｂ３ (R ４ｂ３ is C １－６ represents alkyl. is a nucleoside unit represented by Nucleoside units constituting the compound [B] may be the same or different and have the following general formulas [4e] to [4h]: 【Chemistry 12】 [In the formula, d, B Ｐ , R ４ａ , R ４ｂ１ and R ４ｂ２ has the same meaning as above; ***teeth, (1) the bonding position of the phosphorus atom of the phosphorus bond that is bonded to the oxygen atom at the 5' position of the adjacent nucleoside unit; (2) the bonding position to the substituent [1] or (3) represents the bonding position to a group that can be removed under acidic conditions; and ****teeth, (1) the bonding position of the phosphorus atom of the phosphorus bond that is bonded to the 3'-position oxygen atom or 3'-position nitrogen atom of the adjacent nucleoside unit; (2) the bonding position to the substituent [1] or (3) represents the bonding position to the group that can be removed under acidic conditions; The substituent [1] is represented by the following general formula [1A]: 【Chemistry 13】 (In the formula, ** represents the bond position; D is halogen, 5- to 6-membered saturated cyclic amino or di(C １－６ (alkyl)amino; W represents a lone pair of electrons, 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 that can be removed under basic conditions １－６ alkyl)amino, di(amino-C substituted with a group that can be removed under basic conditions １－６ alkyl)amino or the following general formula [2]: 【Chemistry 14】 (In the formula, * indicates the bonding position with P; a represents an integer of 0 to 2; E is CH ２ , CH-A １ or N-A ２ represents; A １ is C １－６ Alkyl, mono-substituted with a group that can be removed 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 which can be removed under basic conditions, mono (C １－６ alkyl)amino, di(C １－６ alkyl)amino, tri(C １－６ alkyl)ammonio, amino substituted with amidino substituted with a group removable under basic conditions, or the following general formula [3]: 【Chemistry 15】 (In the formula, * represents the bonding position with E; b represents an integer of 0 to 2; c represents 0 or 1; R １１ is C １－６ represents alkyl; and M is CH ２ , an oxygen atom, a sulfur atom, or N-(a group that can be removed under basic conditions). represents a substituent represented by the formula: A ２ is C １－６ Alkyl, mono-substituted with a group that can be removed 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 alkyl, a group that can be removed under basic conditions, aryl, or heteroaryl. represents a substituent represented by the following formula: represents a substituent represented by the following formula: The method according to claim 6, wherein the nucleoside unit is represented by the formula:

8. The nucleoside units constituting the compound [A] may be the same or different and have the following general formula [4d]: 【Chemistry 16】 [In the formula, B Ｐ , * and ** have the same meanings as above. A nucleoside unit represented by The nucleoside units constituting the compound [B] may be the same or different and have the following general formula [4h]: 【Chemistry 17】 [In the formula, B Ｐ , *** and **** have the same meanings as above. The method according to claim 7, wherein the nucleoside unit is represented by the formula:

9. The phosphorus bonds between the nucleoside units constituting the compound [A] and the compound [B] may be the same or different and are represented by the following general formula [5]: 【Chemistry 18】 [In the formula, one of * and ** represents the bonding position to the nitrogen atom at the 3' position of a nucleoside unit, and the other represents the bonding position to the oxygen atom at the 5' position of a nucleoside unit different from said nucleoside unit; W represents 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 that can be removed under basic conditions １－６ alkyl)amino, di(amino-C substituted with a group that can be removed under basic conditions １－６ alkyl)amino or the following general formula [2]: 【Chemistry 19】 (In the formula, * indicates the bonding position with P; a represents an integer of 0 to 2; E is CH ２ , CH-A １ or N-A ２ represents; A １ is C １－６ Alkyl, mono-substituted with a group that can be removed 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 which can be removed under basic conditions, mono (C １－６ alkyl)amino, di(C １－６ alkyl)amino, tri(C １－６ alkyl)ammonio, amino substituted with amidino substituted with a group removable under basic conditions, or the following general formula [3]: 【Chemistry 20】 (In the formula, * represents the bonding position with E; b represents an integer of 0 to 2; c represents 0 or 1; R １１ is C １－６ represents alkyl; and M is CH ２ , an oxygen atom, a sulfur atom, or N-(a group that can be removed under basic conditions). represents a substituent represented by the formula: A ２ is C １－６ Alkyl, mono-substituted with a group that can be removed 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 alkyl, a group that can be removed under basic conditions, aryl, or heteroaryl. represents a substituent represented by the following formula: The method according to any one of claims 6 to 8, wherein the bond is represented by:

10. The compound [B] is represented by the following general formula [B-1]: 【Chemical 21】 [In the formula, B Ｐ are the same or different and have the same meanings as above; D has the same meaning as above; p represents an integer from 1 to 10; Q １ represents a group removable under acidic conditions; W may be the same or different and represent an oxygen atom or a sulfur atom; and X's are the same or different and have the same meanings as defined above. The compound [A] is a compound represented by the following general formula [A-1]: 【Chemical 22】 [In the formula, B Ｐ , W, X, G, and T are as defined above; n represents an integer of 1 to 50. is a compound represented by The compound [C] is a compound represented by the general formula [C-1] 【Chemical 23】 [In the formula, B Ｐ , Q １ , W, X, G, T, n and p are as defined above.] The method according to any one of claims 6 to 9, wherein the compound is represented by the formula:

11. Furthermore, by adding a solution containing an acid to a reaction mixture containing the compound [C-1] produced by the method according to claim 10, Q １ The elimination reactions of the following general formula [E-1] are carried out continuously in the same system to obtain a compound represented by the following general formula [E-1]: 【Chemistry 24】 [In the formula, n, p, B Ｐ , G, T, W and X are as defined above.

11. The method of claim 10, comprising forming a compound represented by the formula:

12. in the presence of at least one reaction accelerator 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; General formula [A-1-1]: 【Chemistry 25】 [In the formula, B Ｐ , Q １ , W, X, G, T and n are as defined above; n is 1 to 25. From the compound Q １ Remove the General formula [A-1]: 【Chemical 26】 [In the formula, B Ｐ , W, X, G, T, and n are as defined above.] and forming a compound of the formula [A-1]. General formula [B-1]: 【Chemical 27】 [In the formula, B Ｐ , Q １ , W and X are as defined above; D is a halogen; and p is an integer from 1 to 10. and reacting the compound General formula [C-1]: 【Chemical 28】 [In the formula, n, p, B Ｐ , Q １ , W, X, G, and T are as defined above.] 12. The method according to any one of claims 1 to 11, for preparing a compound of formula:

13. 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, 1-ethylpyridinium bromide, 1-ethylpyridinium chloride, 1-butylpyridinium chloride, 1-ethyl-3-methylimidazolium chloride, 1-methyl-3-N-octylimidazolium chloride, and 1-butyl-1-methylpyrrolidinium chloride and at least one selected from the group consisting of The amount of the reaction accelerator is in the range of 1 to 100 times by molar ratio relative to 1 mole of the compound of the general formula [A-1]. The method of claim 12.

14. General formula [A-1-1]: 【Chemical 29】 [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 [0033] including desorbing 14. The method of claim 12 or 13.

15. 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 flow reactor to form a compound of the general formula [C-1], and optionally A solution containing the compound of the general formula [C-1] and a solution containing an acid are supplied to a flow reactor, and Q １ to form a compound of formula [E-1], The method according to any one of claims 12 to 14.

16. A solution containing the compound of the general formula [A-1-1] and a solution containing an acid are supplied to a flow reactor, and Q １ to form a compound of formula [A-1], and supplying a solution containing the compound of general formula [A-1] and a solution containing the compound of general formula [B-1] to a subsequent flow reactor to form a compound of general formula [C-1]; The method according to any one of claims 12 to 15.

17. General formula [A-1-2]: 【Chemistry 30】 [In the formula, B Ｐ is an optionally protected nucleobase; Q ２ is H or a group that can be removed under acidic conditions; W is an oxygen atom; X is di(C alkyl)amino; G is a group having the following formula: 【Chemical 31】 [In the formula, * represents the bonding position with T; T is a single bond; n is 1 to 25. Compound.

18. The method according to any one of claims 1 to 16 or the compound according to claim 17, wherein the optionally protected nucleobases are each independently adenine, guanine, hypoxanthine, cytosine, thymine, uracil, or modified bases thereof.

19. The protected nucleobase is a nucleobase in which the amino group or hydroxyl group of the nucleobase is protected with a protecting group, and the protecting groups for the amino group are 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 for the hydroxyl group are each independently selected from the group consisting of 2-cyanoethyl, 4-nitrophenethyl, phenylsulfonylethyl, methyl 18. The method of any one of claims 1 to 16 or the compound of claim 17, wherein the aryl group is selected from the group consisting of phenylsulfonylethyl, 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.

20. A reaction promoter for oligonucleic acid synthesis, comprising, as an active ingredient, 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.

21. 21. The reaction-accelerating agent for oligonucleic acid synthesis according to claim 20, comprising, as an active ingredient, at least one selected from the group consisting of quaternary ammonium salts, quaternary imidazolium salts, and quaternary pyrrolidinium salts.

22. The active ingredient is Tetra C １－１８ Alkylammonium chloride, 1-C １－１８ Alkyl-1-C １－１８ Alkylpyrrolidinium chloride, 1-C １－１８ Alkyl-3-C １－１８ The reaction promoter for oligonucleic acid synthesis according to claim 20, comprising at least one selected from alkylimidazolium chlorides.

23. 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, 1-ethylpyridinium bromide, 1-ethylpyridinium chloride, 1-butylpyridinium chloride, 1-ethyl-3-methylimidazolium chloride, 1-methyl-3-N-octylimidazolium chloride, and 1-butyl-1-methylpyrrolidinium chloride The reaction-promoting agent for oligonucleic acid synthesis according to claim 20, comprising at least one selected from the group consisting of:

24. The reaction promoter for oligonucleic acid synthesis according to claim 20, comprising as an active ingredient at least one selected from the group consisting of tetrabutylammonium chloride, 1-butyl-1-methylpyrrolidinium chloride, and 1-methyl-3-N-octylimidazolium chloride.

25. The reaction promoter for oligonucleic acid synthesis according to any one of claims 20 to 24, for use in the method according to any one of claims 1 to 16.

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