Method for producing oligonucleic acid compound

Abstract

To provide a novel production method that can shorten the production period of an oligonucleic acid compound.SOLUTION: The present invention relates to a method for producing a compound of general formula [C-1-1], which comprises the steps of the formula in the figure. A condensation reaction proceeds efficiently by forming a trivalent phosphorous bond in a condensation reaction of an oligonucleic acid compound.SELECTED DRAWING: Figure 1

Description

Technical Field

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

Background Art

[0002] As methods for producing oligo nucleic acid compounds, a solid-phase method and a liquid-phase method are known. The solid-phase method is a heterogeneous reaction method in which nucleic acids are elongated while bringing a substrate supported on a solid-phase carrier into contact with a solution containing a reaction reagent. In the solid-phase method, a so-called batch method is used in which a reaction is carried out in a reaction vessel with a filter (see, for example, Non-Patent Document 1 and Patent Document 1). Also known is a pseudo-flow synthesis method in which a solid-phase carrier is placed in a column and a solution containing a reaction reagent is passed through the column to cause a reaction, such as a nucleic acid automatic synthesizer (for example, a DNA or RNA synthesizer). On the other hand, the liquid-phase method is a homogeneous reaction method in which nucleic acids are elongated by reacting in a solution containing both a substrate and a reaction reagent. The liquid-phase method also uses a batch method in which a reaction is carried out in a container (see, for example, Patent Document 2 and Patent Document 3).

[0003] In any case of the solid-phase method, the liquid-phase method, the batch method, and the pseudo-flow synthesis method, in the chemical synthesis method of oligo nucleic acid compounds, a "deprotection" reaction for removing a protecting group of an oxygen atom or an amino group on the nucleic acid compound, and the formation of a bond between an oxygen atom or a nitrogen atom that has been deprotected and can react and a phosphorus atom are repeatedly carried out many times to elongate the nucleic acid. Among them, controlling the reaction efficiency and reaction rate in the "condensation" reaction for forming a bond between a phosphorus atom and an oxygen atom or a nitrogen atom is very important in the production of oligo nucleic acid compounds, and the conditions of this condensation reaction are factors that have a great impact on the production period of oligo nucleic acid compounds.

[0004] Since the solid-phase method is a heterogeneous reaction between a solid support and a solution, it is known that the reactivity of the condensation reaction decreases due to steric hindrance caused by the solid support. Generally, polystyrene resin is used as the solid support. During the reaction, it swells in the reaction solvent used and its volume becomes larger than in the dry state. The degree of swelling depends on the reaction solvent. Therefore, the reaction efficiency and reaction rate of the condensation reaction in the solid-phase method depend on the reaction solvent used. In particular, polystyrene resin does not swell significantly in polar solvents such as acetonitrile, which is commonly used for the synthesis of oligonucleic acid compounds. Using a polar solvent in the solid-phase method is not necessarily favorable in terms of improving the reaction efficiency and reaction rate of the condensation reaction.

[0005] On the other hand, as homogeneous reaction methods, a liquid-phase method and a synthesis method using hydrophobic group-linked nucleosides, pseudo-solid-phase protected nucleosides, etc. are known. The liquid-phase method is a homogeneous reaction method in which both the substrate and the reaction reagent are reacted in a solution. Compared with the solid-phase method, the reaction efficiency is high and the reaction rate is fast. However, in order to remove the reaction reagent and reaction solvent that become impurities, column purification etc. are required. The synthesis method using hydrophobic group-linked nucleosides, pseudo-solid-phase protected nucleosides, etc. can react in a homogeneous system like the liquid-phase method. Therefore, compared with the solid-phase method, the reaction efficiency is high and the reaction rate is fast. Furthermore, after the reaction, unnecessary reaction reagents and reaction solvents can be removed by precipitating the target compound from the reaction mixture (for example, see Patent Document 4). In these homogeneous reaction methods, non-polar solvents such as chloroform are used in the condensation reaction. However, as reported in the synthesis of morpholino nucleic acids (for example, see Patent Document 5), the condensation reaction in a non-polar solvent requires a very long time. Therefore, using a non-polar solvent in a homogeneous reaction is not necessarily favorable in terms of improving the reaction efficiency and reaction rate of the condensation reaction.

Prior Art Documents

Patent Documents

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

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

[0008] An object of the present invention is to provide a novel production method capable of shortening the production period of an oligo nucleic acid compound. [Means for Solving the Problems]

[0009] The present inventors have found that in the condensation reaction of an oligo nucleic acid compound, the condensation reaction proceeds efficiently by forming a trivalent phosphorus bond, and thus completed the present invention.

[0010] The present invention relates to, for example, a compound [A] having a hydroxyl group or a primary or secondary amino group and the following general formula [1]: [Chemical Formula] [In the formula, * represents the bonding position with the residue of compound [B]; D represents a halogen, a 5 - 6 membered saturated cyclic amino group or di(C 1-6 alkyl)amino; W 0represents a lone pair of electrons, an oxygen atom or a sulfur atom; and X is a hydroxyl group substituted with a group capable of leaving 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 1-6 alkyl)amino substituted with a group capable of leaving under basic conditions, di(amino-C 1-6 alkyl)amino substituted with a group capable of leaving under basic conditions or the following general formula [2]:

Chemical formula

Chemical formula

Chemical formula

Advantages of the Invention

[0011] An oligonucleic acid compound is a compound having a structure in which two or more nucleoside units are linked via a phosphorus bond. In order to produce an oligonucleic acid compound, it is necessary to perform a condensation reaction many times to form a phosphorus bond between adjacent nucleoside units. According to the present invention, since a phosphorus bond can be efficiently formed, it can be expected that the production time of the oligonucleic acid compound will be shortened as a result.

Brief Description of the Drawings

[0012]

Figure 1

Embodiments for Carrying Out the Invention

[0013] The present invention is a method for producing a compound [C] by subjecting a compound [A] having a hydroxyl group or a primary or secondary amino group and a compound [B] having a substituent [1] to a condensation reaction, characterized in that a trivalent phosphorus bond is formed by the condensation reaction.

[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] One specific embodiment of the compound [A] can include a compound containing one to a plurality of nucleoside units in its molecule. Specifically, a compound containing nucleoside units in the range of one to 50 is suitable, a compound containing nucleoside units in the range of one to 30 is preferred, and a compound containing nucleoside units in the range of one to 25 is more preferred. Examples of the nucleoside units contained in the compound [A] include the following general formulas [4a] to [4d]:

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[10] :

Chemical formula

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

[11] :

Chemical formula

[11] "), provided that when G is a silicon substituent, T is a single bond.) represents the bonding position with a substituent (hereinafter referred to as "substituent [6]") represented by; ** is (1) the bonding position with a phosphorus bond that is bonded to an oxygen atom or a nitrogen atom at the 3'-position of an adjacent nucleoside unit, (2) the bonding position with a hydrogen atom or (3) the bonding position with substituent [6]; 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 eliminated under neutral conditions, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkoxy-C 1-6 alkyl, halogen, nitro or cyano; R 4b1 and R 4b2 each independently represent a hydrogen atom or C 1-6 alkyl, or R 4b1 and R 4b2 together with the adjacent carbon atom form a carbonyl; and J is an oxygen atom or N-R 4b3 (R 4b3 is C 1-6represents alkyl.) represents.] Nucleoside units represented by (hereinafter referred to as "nucleoside unit [4a]", "nucleoside unit [4b]", "nucleoside unit [4c]", "nucleoside unit [4d]"), respectively, can be mentioned.

[0016] Preferred embodiments of nucleoside units [4a] to [4d] are, for example, the following general formulas [4a1] to [4d1], respectively: [Chemical formula] [In the formula, d, B P , J, R 4a , R 4b1 and R 4b2 have the same meanings as described above; * represents (1) the bonding position with the phosphorus bond bonded to the oxygen atom at the 5'-position of the adjacent nucleoside unit, (2) the bonding position with a hydrogen atom or (3) the bonding position with substituent [6]; and ** represents (1) the bonding position with the phosphorus bond bonded to the oxygen atom or nitrogen atom at the 3'-position of the adjacent nucleoside unit, (2) the bonding position with a hydrogen atom or (3) the bonding position with substituent [6].] Nucleoside units represented by (hereinafter referred to as "nucleoside unit [4a1]", "nucleoside unit [4b1]", "nucleoside unit [4c1]", "nucleoside unit [4d1]"), respectively, can be mentioned.

[0017] When compound [A] contains a plurality of nucleoside units in its molecule, it is preferable that the adjacent nucleoside units in the compound are bonded to each other via a phosphorus bond. As the phosphorus bond between each nucleoside unit constituting compound [A], they can be the same or different, and for example, the following general formula [5]: [Chemical formula] [In the formula, X has the same meaning as described above; One of * and ** represents the bonding position with the oxygen atom or nitrogen atom at the 3'-position of the nucleoside unit, and the other represents the bonding position with the oxygen atom at the 5'-position of a nucleoside unit different from the said nucleoside unit; and W represents a lone pair, an oxygen atom or a sulfur atom.] The bond represented by (hereinafter referred to as "phosphorus bond [5]") can be mentioned. W preferably represents a lone pair or an oxygen 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 a plurality of nucleoside units [4d] In the nucleoside unit represented by the general formula [4d], * is (1) The bonding position with the phosphorus bond bonded to the oxygen atom at the 5'-position of the adjacent nucleoside unit or (2) The bonding position with a hydrogen atom; ** is (1) The bonding position with the phosphorus bond bonded to the nitrogen atom at the 3'-position of the adjacent nucleoside unit or (2) The bonding position with substituent [6].

[0020] As one of the embodiments of compound [A], for example, a compound in which the oxygen atom at the 5'-position of the nucleoside unit on the 5'-terminal side is substituted with, for example, substituent [6] can be mentioned. In that case, examples of the phosphorus bond between each nucleoside unit constituting the compound [A] include, for example, the phosphorus bond [5], which may be the same or different. However, in the phosphorus bond represented by the general formula [5], one of * and ** represents the bonding position with the nitrogen atom at the 3'-position of the nucleoside unit, and the other represents the bonding position with the oxygen atom at the 5'-position of a nucleoside unit different from the said nucleoside unit.

[0021] As a more specific embodiment of the compound [A], for example, the following general formula [A-1]:

Chemical formula

Chemical formula

Chemical formula

[0022] (A-2) Compound [A] composed of one to a plurality of 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 formulas [4a], [4b] and [4c], * is (1) The bonding position with the phosphorus bond bonded to the oxygen atom at the 5'-position of the adjacent nucleoside unit or (2) The bonding position with the substituent [6]; ** is (1) The bonding position with the phosphorus bond bonded to the oxygen atom at the 3'-position of the adjacent nucleoside unit or (2) The bonding position with a hydrogen atom.

[0023] As one aspect of compound [A], for example, a compound in which the oxygen atom at the 3'-position of the nucleoside unit on the 3'-terminal side is substituted with, for example, substituent [6] can be mentioned. In that case, the phosphorus bond between each nucleoside unit constituting compound [A] is, for example, suitable for phosphorus bond [5]. However, in the phosphorus bond represented by the general formula [5], one of * and ** represents the oxygen atom at the 3'-position of the nucleoside unit, and the other represents the bonding position with the oxygen atom at the 5'-position of a nucleoside unit different from the said nucleoside unit. As a more specific aspect of compound [A], for example, the following general formula [A-2]:

Chemical formula

[0024] Specific examples of the substituent [7] in compound [A-1] and compound [A-2] include, for example, the following substituents.

Chemical Formula

[0025] (B) Regarding compound [B] Examples of compound [B] that can be used in this production method include, for example, compounds having substituent [1].

[0026] One specific embodiment of compound [B] includes, for example, compounds containing one to a plurality of nucleoside units in their molecules. More specifically, compounds containing nucleoside units in the range of one to ten are suitable, compounds containing nucleoside units in the range of one to seven are preferred, and compounds containing nucleoside units in the range of one to five are more preferred.

[0027] Examples of the nucleoside units contained in compound [B] include the following general formulas [4e] to [4h]:

Chemical Formula

[0028] Preferred embodiments of the nucleoside units [4e] to [4h] are, for example, the following general formulas [4e1] to [4h1]:

Chemical formula

[0029] When compound [B] contains a plurality of nucleoside units in its molecule, it is preferable that the adjacent nucleoside units in the compound are bonded to each other via a phosphorus bond. In that case, examples of the phosphodiester bond between each nucleoside unit constituting compound [B] may include, for example, phosphodiester bond [5], which may be the same or different from each other. However, in the phosphodiester bond represented by the general formula [5], one of * and ** represents an oxygen atom at the 3'-position of the nucleoside unit, and the other represents the bonding position with an oxygen atom at the 5'-position of a nucleoside unit different from the said nucleoside unit.

[0030] Examples of compound [B] are shown below.

[0031] (B-1) Compound [B] composed of one to a plurality of nucleoside units [4h] In the nucleoside unit represented by the general formula [4h], *** is (1) the bonding position with a phosphodiester bond that binds to an oxygen atom at the 5'-position of an adjacent nucleoside unit or (2) the bonding position with a group that can be eliminated under acidic conditions; **** is (1) the bonding position with a phosphodiester bond that binds to a nitrogen atom at the 3'-position of an adjacent nucleoside unit or (2) the bonding position with substituent [1].

[0032] As one of the embodiments of compound [B], 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 eliminated under acidic conditions can be mentioned. In that case, examples of the phosphodiester bond between each nucleoside unit constituting compound [B] may include, for example, phosphodiester bond [5], which may be the same or different from each other. However, in the phosphodiester bond represented by the general formula [5], one of * and ** represents a nitrogen atom at the 3'-position of the nucleoside unit, and the other represents the bonding position with an oxygen atom at the 5'-position of a nucleoside unit different from the said nucleoside unit. In addition, 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]:

Chemical formula

Chemical formula

[0033] As a specific example of compound [B-1] where p = 1, for example, the compounds described in Table 1 below can be mentioned.

[0034]

Table : 1

[0035] (B-2) Compound [B] composed of one to a plurality of 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 formulas [4e], [4f] and [4g], *** is (1) The bonding position with the phosphorus bond bonded to the oxygen atom at the 3'-position of the adjacent nucleoside unit or (2) The bonding position with the substituent [1]; **** is (1) The bonding position with the phosphorus bond bonded to the oxygen atom at the 5'-position of the adjacent nucleoside unit or (2) The bonding position with a group eliminable under acidic conditions.

[0036] As one aspect of compound [B], for example, a compound can be mentioned in which the oxygen atom at the 5'-position of the nucleoside unit on the 5'-terminal side is substituted with a group that can be eliminated under acidic conditions. In this case, as the phosphodiester bond between each nucleoside unit constituting compound [B], for example, the phosphodiester bond [5] can be mentioned, which may be the same or different from each other. However, in the phosphodiester bond represented by the general formula [5], one of * and ** represents the oxygen atom at the 3'-position of the nucleoside unit, and the other represents the bonding position with the oxygen atom at the 5'-position of a nucleoside unit different from the said nucleoside unit. In addition, the oxygen atom at the 3'-position of the nucleoside unit on the 3'-terminal side of compound [B] preferably has a substituent containing a phosphorus atom represented by the following general formula [1B]: [Chemical formula] [wherein, D and X have the same meanings as described above; and ** represents the bonding position with the residue of compound [B].] It is appropriate to have a substituent containing a phosphorus atom represented by the formula. As a more specific aspect of compound [B], for example, the following general formula [B-2]: [Chemical formula] [wherein, p, B P , D, Q 1 , R 4a , X and W have the same meanings as described above.] The compound represented by the formula (hereinafter referred to as "compound [B-2]") can be mentioned.

[0037] As specific examples of compound [B-2] where p = 1, for example, the compounds described in Table 2 below can be mentioned. In Table 2, DMTr represents dimethoxytrityl, and TBDMS represents tert-butyldimethylsilyl.

[0038] [Table 2]

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

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

[0041] (C-1) Compound [C] composed of one to a plurality of nucleoside units [4d] and one to a plurality of nucleoside units [4h] As a specific embodiment of compound [C], for example, the following general formula [C-1]:

Chemical formula

Chemical formula

[0042] Compound [B-1-1] has the general formula:

Chemical formula

[0043] When p = 1, specific examples of compound [B-1-1] include, for example, the compounds listed in Table 3 below.

[0044]

Table 3

[0045] Compound [B-1-2] has the general formula:

Chemical formula

[0046] When p = 1, specific examples of compound [B-1-2] include, for example, the compounds listed in Table 4 below.

Table 4

[0047] (C-2) Compound [C] composed of one to a plurality of nucleoside units selected from the group consisting of nucleoside unit [4a], nucleoside unit [4b], and nucleoside unit [4c] and one to a plurality of nucleoside units selected from the group consisting of nucleoside unit [4e], nucleoside unit [4f], and nucleoside unit [4g]

[0048] As one of the embodiments of the specific compound [C], for example, the following general formula [C-2]: [Chemical formula] [In the formula, n, p, B P , G, Q 1 , R 4a , T, W and X have the same meanings as described above.] The compound represented by (hereinafter referred to as "compound [C-2]") can be mentioned. As will be described later, in the method for producing compound [C-2] by reacting compound [A-2] and compound [B-2], as the phosphorus bond newly formed, for example, the following general formula [5a]: [Chemical formula] [In the formula, X has the same meaning as described above; and One of * and ** represents the bonding position with the oxygen atom at the 3'-position of the nucleoside unit, and the other represents the bonding position with the oxygen atom at the 5'-position of a nucleoside unit different from the said nucleoside unit.] The bond containing a phosphorus atom represented by (hereinafter referred to as "phosphorus bond [5a]") can be mentioned.

[0049] By reacting compound [C-2] with an oxidizing agent, the phosphorus atom on the phosphorus bond in its molecule is oxidized to the following general formula [D-2]: [Chemical formula] [In the formula, n, p, B P , G, Q 1 , R 4a , T, W and X have the same meanings as described above.] It can be converted into a compound represented by (referred to as "Compound [D-2]").

[0050] (D) Explanation of terms Here, examples of the "nucleic acid base" include adenine, guanine, hypoxanthine, cytosine, thymine, uracil, or modified bases thereof. Such modified bases include, for example, 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, xanthine, but are not limited thereto. However, the amino group or hydroxyl group of the nucleic acid base related to B P may be protected. As used herein, the "optionally protected nucleic acid base" includes both unprotected "nucleic acid bases" and protected "nucleic acid bases", and examples include adenine, guanine, hypoxanthine, cytosine, thymine, uracil, etc., in which the amino group and / or hydroxyl group is unprotected or protected. The protecting group for the amino group is not particularly limited as long as it can be used as a protecting group for nucleic acids. Specifically, for example, benzoyl, 4-methoxybenzoyl, acetyl, propionyl, butyryl, isobutyryl, phenylacetyl, phenoxyacetyl, 4-tert-butylphenoxyacetyl, 4-isopropylphenoxyacetyl, (dimethylamino)methylene can be mentioned. As the protecting group for the amino group, benzoyl, acetyl, phenylacetyl, 4-tert-butylphenoxyacetyl are preferable. As the protecting group for the hydroxyl group, for example, 2-cyanoethyl, 4-nitrophenethyl, phenylsulfonylethyl, methylsulfonylethyl, trimethylsilylethyl, phenyl which may be substituted with 1 to 5 electron-withdrawing groups at any substitutable position, diphenylcarbamoyl, dimethylcarbamoyl, diethylcarbamoyl, methylphenylcarbamoyl, 1-pyrrolidinylcarbamoyl, morpholinocarbamoyl, 4-(tert-butylcarboxy)benzyl, 4-[(dimethylamino)carboxy]benzyl, 4-(phenylcarboxy)benzyl can be mentioned (see, for example, International Publication No. 2009 / 064471A1). As the protecting group for the hydroxyl group, 2-cyanoethyl, 4-nitrophenethyl, 4-(tert-butylcarboxy)benzyl are preferable. The protecting group for the 6-position hydroxyl group of guanine is preferably 2-cyanoethyl. In one embodiment, examples of the protected nucleobases include those shown below. [Chemical formula] [In the formula, Pg represents a protecting group.] More specific embodiments of the protected nucleobases include adenine (A Bz ) in which the amino group is protected with benzoyl, cytosine (C Bz ) in which the amino group is protected with benzoyl, guanine (G CE,Pac ) in which the hydroxyl group is protected with 2-cyanoethyl and the amino group is protected with phenoxyacetyl, etc., but are not limited thereto. "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. Examples of the "long-chain alkyl" moiety in "long-chain alkyl-carbonyl" and "long-chain alkyloxy" may be the same as those of the above-mentioned "long-chain alkyl". "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. Examples of the "long-chain alkenyl" moiety in "long-chain alkenyloxy" and "long-chain alkenyl-carbonyl" may be the same as those of the above-mentioned "long-chain alkenyl". "Halogen" can include, for example, a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom. Examples of the "5- to 6-membered saturated cyclic amino" include 5- to 6-membered saturated cyclic amino groups having one or two N atoms and optionally one O or S atom as ring-constituting atoms, specifically, 1-pyrrolidinyl, 1-imidazolidinyl, piperidino, 1-piperazinyl, 1-tetrahydropyrimidinyl, 4-morpholino, 4-thiomorpholino, 1-homopiperazinyl, and oxazolidin-3-yl. "C 1-6 alkyl" refers to a linear or branched alkyl having 1 to 6 carbon atoms, specifically, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl. "C 1-6 alkoxy" refers to a linear or branched alkoxy having 1 to 6 carbon atoms, specifically, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentyloxy, and n-hexyloxy. "C 1-6 alkoxy-C1-6 The "C" of "alkyl" 1-6 The "alkoxy" part can be the same as the above-mentioned "C 1-6 "alkoxy". "Di(C 1-6 alkyl)amino", mono(amino-C 1-6 alkyl)amino substituted with a group capable of leaving under basic conditions, di(amino-C 1-6 alkyl)amino substituted with a group capable of leaving under basic conditions, 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 of "C 1-6 alkyl" part can be the same as the above-mentioned "C 1-6 alkyl". "C 2-10 "alkylene" is a divalent group formed by removing one hydrogen atom bonded to different constituent carbon atoms from a linear or branched alkyl having 2 to 10 carbon atoms. For example, an ethylene group, a propylene group, an isopropylene group, a butylene group, a pentylene group, a hexylene group can be mentioned. Such "alkylene" may be substituted with 1 to 12 halogens at any substitutable position. L 1 As the "alkylene" related to L, ethylene is particularly preferred. "C 6-10 "arylene" is a divalent group formed by removing two hydrogen atoms bonded to two different ring-constituting carbon atoms from a monocyclic or polycyclic aromatic hydrocarbon having 6 to 10 carbon atoms. For example, phenylene, naphthylene can be mentioned. Such "arylene" may be substituted with 1 to 6 halogens at any substitutable position. L1 As the "arylene" according to [0], 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 1-6 alkyl)amino substituted with a group eliminable under basic conditions", "mono(C 1-6 alkyl)amino-C 1-6 alkyl substituted with a group eliminable under basic conditions", "mono(amino-C 1-6 alkyl substituted with a group eliminable under basic conditions)amino", "di(amino-C 1-6 alkyl substituted with a group eliminable under basic conditions)amino", the "C 1-6 alkyl" part can be the same as the above-mentioned "C 1-6 alkyl". Examples of the "group eliminable under acidic conditions" include trityl, monomethoxytrityl, tert-butyldimethylsilyl, and dimethoxytrityl. Examples of the "group eliminable under basic conditions" include trifluoroacetyl. Examples of the "group eliminable under neutral conditions" include groups that can be eliminated by the action of tetrabutylammonium fluoride or hydrogen fluoride·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. "Heteroaryl" can include, for example, pyridyl, pyrimidyl, pyridazyl, pyrazinyl, thienyl, furanyl. As the "solid support", generally, any material that can be used for solid-phase synthesis of nucleic acids, peptides, peptide nucleic acids, sugars, etc. can be used without particular problem. Examples include controlled pore glass (CPG), oxalylated-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 - copolymer of polystyrene / divinylbenzene, polystyrene resin, polyacrylamide resin. Examples of the "soluble polymer soluble in an organic solvent" can include a styrene polymer without crosslinking and a polyethylene glycol derivative. For the "soluble polymer soluble in an organic solvent" part of "(soluble polymer soluble in an organic solvent)-oxy" and "(soluble polymer soluble in an organic solvent)-amino", the same ones as the above-mentioned "soluble polymer soluble in an organic solvent" can be mentioned. Examples of the "styrene polymer without crosslinking" can include derivatives having a spacer such as polyethylene glycol in polystyrene not crosslinked with divinylbenzene (TentaGel series, ArgoGel series). Examples of the "polyethylene glycol derivative" can include derivatives having a substituent with polyethylene glycol having a molecular weight of 100 to 40000 (SUNBRIGHT (registered trademark) series).

[0051] (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 and compound [B] having substituent [1] to a condensation reaction. As described in the examples and test examples to be described later, by forming a trivalent phosphorus bond in the production of compound [C], the condensation reaction can proceed efficiently.

[0052] 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 art, and a single solvent may be used, or two or more solvents may be mixed and used. Examples of the solvent 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; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, and cyclohexane, and halogenated solvents. These solvents may be used in combination. Examples of the halogenated solvent that can be used in this production method include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, or a mixed solvent thereof. Among them, chloroform, dichloromethane, 1,1-dichloroethane, and 1,2-dichloroethane are preferred. In this production method, a base may be used if necessary. Examples of the "base" that can be used in this production method include diisopropylamine, N,N-diisopropylethylamine, triethylamine, N-ethylmorpholine, and 2,6-lutidine. The amount of the base used in this production method is, for example, appropriately in the range of 1 to 100 times the molar ratio with respect to 1 mol of compound [A], preferably in the range of 1 to 10 times, and more preferably in the range of 1 to 5 times. In this production method, an additive may be used if necessary. Examples of the "additive" that can be used in this production method include LiBr, LiCl, LiI, and NaI. As the amount of the additive that can be used in this production method, for example, within the range of 0.2 to 6.0 times the molar amount, preferably within the range of 0.4 to 3.0 times the molar amount, and more preferably within the range of 1.0 to 2.5 times the molar amount, based on 1 mol of compound [A], is appropriate. As the reaction temperature, for example, within the range of -78°C to 130°C is appropriate, 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] used, the type of compound [B], the type of reaction solvent, the type of base, and the reaction temperature. For example, within the range of 1 minute to 300 minutes is appropriate, and preferably within the range of 5 minutes to 120 minutes.

[0053] When it is possible to produce compound [C] which is an oligonucleic acid compound, this production method can be applied either by the batch method or the flow method. Furthermore, this production method can also be applied in the solid-phase method, the liquid-phase method, and the liquid-phase method using a hydrophobic group-bonded nucleoside or a pseudo-solid-phase protected nucleoside, etc., which are known as production methods of oligonucleic acid compounds.

[0054] When it is possible to produce compound [C] which is an oligonucleic acid compound using the solid-phase method, those supported on a solid-phase carrier can be used at the oxygen atom at the 3'-position of the nucleoside unit on the 3'-terminal side of compound [A] or at the oxygen atom at the 5'-position of the nucleoside unit on the 5'-terminal side of compound [A].

[0055] When it is possible to produce an oligonucleic acid compound using the liquid-phase method, those supported on a soluble polymer dissolved in an organic solvent can be used at the oxygen atom at the 3'-position of the nucleoside unit on the 3'-terminal side of compound [A] or at the oxygen atom at the 5'-position of the nucleoside unit on the 5'-terminal side of compound [A].

[0056] When an oligonucleic acid compound can be produced by using a liquid phase method using a hydrophobic group-bonded nucleoside, a pseudo-solid phase-protected nucleoside, or the like, at the oxygen atom at the 3'-position of the nucleoside unit on the 3'-terminal side of compound [A] or at the oxygen atom at the 5'-position of the nucleoside unit on the 5'-terminal side of compound [A], for example, those having a hydrophobic group bonded thereto or those supported on a pseudo-solid phase can be used (see, for example, JP-A-2010-275254 and WO 2012 / 157723).

[0057] Hereinafter, the compound [C-1] and the compound [C-2] will be described in detail by way of example.

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

[0059] 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 art, and a single solvent may be used, or two or more solvents may be mixed and used. Examples of the solvent 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; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, and cyclohexane, halogenated solvents, nitriles such as acetonitrile and propionitrile, ethers such as THF, 1,4-dioxane, and diethyl ether, amides such as dimethylformamide and dimethylacetamide, and 1,3-dimethyl-2-imidazolidinone. These solvents may be used in combination. Examples of the halogen-based 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, or a mixed solvent thereof. Among these, chloroform, dichloromethane, 1,1-dichloroethane, and 1,2-dichloroethane are preferred. In this production method, a base may be used as necessary. Examples of the "base" that can be used in this production method include diisopropylamine, N,N-diisopropylethylamine, triethylamine, N-ethylmorpholine, and 2,6-lutidine. As for the amount of the base used in this production method, for example, a molar ratio in the range of 1 to 100 times, preferably in the range of 1 to 10 times, and more preferably in the range of 1 to 5 times with respect to 1 mol of compound [A] is appropriate. In this production method, an additive may be used as necessary. Examples of the "additive" that can be used in this production method include LiBr, LiCl, LiI, and NaI, which are preferred. As for the amount of the additive used in this production method, for example, a molar ratio in the range of 0.2 to 6.0 times, preferably in the range of 0.4 to 3.0 times, and more preferably in the range of 1.0 to 2.5 times with respect to 1 mol of compound [A] is appropriate. As for the reaction temperature, for example, a range of -78°C to 130°C is appropriate, a range of -40°C to 100°C is preferred, and a range of 0°C to 80°C is more preferred. The reaction time varies depending on the types of compound [A] and compound [B] used, the type of reaction solvent, the type of base, and the reaction temperature. For example, a range of 1 minute to 300 minutes is appropriate, and a range of 5 minutes to 120 minutes is preferred.

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

[0061] In the compound [A-1], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (4) a substituent [7] (provided that the case where Z is a solid-phase carrier is excluded), for example, (1) stirring the compound [A-1] and the compound [B-1] in a reaction solvent in a suitable reaction vessel, or (2) independently supplying a solution containing the compound [A-1] and a solution containing the compound [B-1] through a supply flow path into a flow-through reactor or a reaction flow path, and mixing these solutions in the flow-through reactor or the like, this condensation reaction can be carried out.

[0062] Here, the "supply flow path" means a flow path for continuously supplying a solution, the "reaction flow path" means a flow path through which a solution can be reacted while flowing, and the flow-through reactor means a reactor that can simultaneously perform the input of a solution, the reaction, and the recovery of the product without interruption of the operation. As means for supplying a solution containing the compound [A-1] and a solution containing the compound [B-1] to the supply flow path, pumps for supplying liquids commonly used in this field can be generally mentioned, specifically, for example, syringe pumps, plunger pumps, diaphragm pumps, gear pumps. Examples of the flow-through reactor include in-line mixers such as microreactors and static mixers. As a means for guiding a solution containing compound [A-1] and a solution containing compound [B-1] from a supply channel to a reaction channel, for example, a multi-stage collision type micromixer can be mentioned. As the material of the supply channel and the reaction channel, for example, a synthetic resin tube 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 a metal tube selected from the group consisting of stainless steel, copper and its alloys, and titanium and its alloys can be mentioned. The inner diameters of the supply channel and the reaction channel may be appropriately selected, for example, from the sizes usually in the range of 0.1 mm to 1.0 mm, but it is preferably selected from the sizes in the range of 0.2 mm to 1.0 mm.

[0063] (E-1-1) Method for producing compound [C-1] As one aspect of the present invention, Step 1) General formula [B-1-1]:

Chemical formula

Chemical formula

Chemical formula

Chem.

Chem.

Chem.

[0064] (E-1-2) Method for producing compound [C-1] As one aspect of the present invention, Step 1') the general formula [A-1]:

Chem.

Chem.

[0065] General formula [P]: [Chemical formula] [wherein X is as defined above, LG 1 and LG 2is the same or different and is a leaving group, for example, a halogen (chloro, bromo, iodo, especially chloro). The compound of ] is hereinafter also referred to as compound [P]. Compound [P] is commercially available or can be prepared by methods commonly used in the art. Examples of compound [P] include dichloro(dimethylamino)phosphine.

[0066] (E-1-1) and reaction conditions of the production method of (E-1-2) Step 1) and Step 1’) In this step, for example, with respect to 1 mole of compound [A-1] or compound [B-1-1], compound [P] is preferably used within a molar ratio range of 0.6 to 4.0 times, and more preferably within a range of 0.75 to 1.5 times. In this step, a base may be used if necessary. Examples of the "base" that can be used in this step include N,N-diisopropylethylamine, triethylamine, 1,8-bis(dimethylamino)naphthalene, pyridine, 2,4,6-collidine, N-ethylmorpholine, diazabicycloundecene (DBU), 1,4-diazabicyclo[2.2.2]octane (DABCO), and N-methylimidazole. Preferably, N,N-diisopropylethylamine, triethylamine, 1,8-bis(dimethylamino)naphthalene, pyridine, 2,4,6-collidine are used. As the amount of the base that can be used in this production method, for example, with respect to 1 mole of compound [A] or compound [B], a molar ratio range of 1 to 10 times is appropriate, and a range of 1 to 5 times is preferred. The solvent that can be used in this step is not particularly limited. For example, dichloromethane, tetrahydrofuran, dimethyl sulfoxide, and mixtures thereof can be used. Dichloromethane, 10% tetrahydrofuran / dichloromethane, 10% dimethyl sulfoxide / dichloromethane are preferred, and dichloromethane, 10% tetrahydrofuran / dichloromethane is more preferred. The reaction temperature of this step is not particularly limited. For example, it is -78 to 60 °C. The reaction time of this step is not particularly limited, but for example, it is 0.05 to 20 minutes, preferably 0.2 to 10 minutes.

[0067] Step 2) and Step 2’) In this step, a base may be used as necessary. Examples of the "base" that can be used in this step include N,N-diisopropylethylamine, triethylamine, 1,8-bis(dimethylamino)naphthalene, pyridine, 2,4,6-collidine, N-ethylmorpholine, diazabicycloundecene (DBU), 1,4-diazabicyclo[2.2.2]octane (DABCO), and N-methylimidazole. Preferably, they are N,N-diisopropylethylamine, triethylamine, 1,8-bis(dimethylamino)naphthalene, pyridine, and 2,4,6-collidine. The amount of the base used in this production method is, for example, suitably in the range of 1 to 10 times the molar ratio, preferably in the range of 1 to 5 times the molar ratio, relative to 1 mol of compound [A-1] or compound [B-1-1]. The solvent that can be used in this step is not particularly limited, but for example, dichloromethane, tetrahydrofuran, 2-methyltetrahydrofuran, dimethyl sulfoxide, and mixtures thereof can be used. Dichloromethane, 10% tetrahydrofuran / dichloromethane, 10% 2-methyltetrahydrofuran / dichloromethane, and 10% dimethyl sulfoxide / dichloromethane are preferred, and dichloromethane, 10% tetrahydrofuran / dichloromethane, and 10% 2-methyltetrahydrofuran / dichloromethane are more preferred. The reaction temperature of this step is not particularly limited, but for example, it is -78 to 60°C, preferably 0 to 60°C. The reaction time of this step is not particularly limited, but for example, it is 0.5 to 60 minutes, preferably 2 to 30 minutes, and more preferably 1 to 30 minutes.

[0068] Step 3) and Step 3’) In this step, for example, with respect to 1 mole of compound [C-1-1], the oxidizing agent is in the range of 1 to 10 times the molar ratio, preferably in the range of 1.5 to 3 times. In this step, as the oxidizing agent, for example, iodine, magnesium monoperoxyphthalate hexahydrate (MMPP), peracetic acid, meta-chloroperbenzoic acid (mCPBA), tert-butyl hydroperoxide (TBHP), N-methylmorpholine N-oxide, hydrogen peroxide, manganese dioxide can be used, and iodine, magnesium monoperoxyphthalate hexahydrate (MMPP) are preferred. The solvent that can be used in this step is not particularly limited. For example, it can be appropriately selected according to the oxidizing agent used. For example, water, chloroethane, dichloromethane, chloroform, tetrahydrofuran, or a mixture thereof can be used. As the solvent that can be used in this step, when iodine is used as the oxidizing agent, for example, dichloromethane, chloroform, tetrahydrofuran, 0.2% water / tetrahydrofuran can be used; when MMPP is used, for example, water can be used; when peracetic acid is used, for example, dichloromethane can be used; when mCPBA is used, for example, dichloromethane can be used; when TBHP is used, for example, water, dichloromethane can be used; when N-methylmorpholine N-oxide is used, dichloromethane can be used; when hydrogen peroxide is used, for example, water can be used; when manganese dioxide is used, for example, dichloromethane can be used. The reaction temperature of this step is not particularly limited. For example, it is 0 to 25 °C. The reaction time of this step is not particularly limited. For example, it is 1 to 60 minutes, preferably 5 to 30 minutes.

[0069] When the compound [A-1] has a solid-phase carrier in its molecule, that is, when G is a substituent [7] and Z is a solid-phase carrier in the compound [A-1], for example, (1) filling a suitable column with the compound [A-1] and eluting a reaction solution containing the compound [B-1-1], or (2) shaking or stirring a reaction solution containing the compound [A-1] and the compound [B-1-1] in a reaction vessel with a filter, the present condensation reaction can be carried out.

[0070] In the compound [A-1], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (4) a substituent [7] (however, excluding the case where Z is a solid-phase carrier), for example, (1) stirring the compound [A-1] and the compound [B-1-1] in a reaction solvent in a suitable reaction vessel, or (2) independently supplying a solution containing the compound [A-1] and a solution containing the compound [B-1-1] through a supply flow path into a flow-through reactor or a reaction flow path, and mixing these solutions in the flow-through reactor or the like, the present condensation reaction can be carried out.

[0071] Here, the "supply flow path" means a flow path for continuously supplying a solution, the "reaction flow path" means a flow path through which a solution can be reacted while flowing, and the flow-through reactor means a reactor that simultaneously performs the input, reaction, and recovery of a product without interruption of the operation. As means for supplying a solution containing the compound [A-1] and a solution containing the compound [B-1-1] to the supply flow path, pumps for supplying liquids usually used in this field can be specifically mentioned, for example, syringe pumps, plunger pumps, diaphragm pumps, and gear pumps. Examples of the flow-through reactor include microreactors and in-line mixers such as static mixers. As means for guiding a solution containing compound [A-1] and a solution containing compound [B-1-1] from a supply channel to a reaction channel, for example, a multi-stage collision type micromixer can be mentioned. As the material of the supply channel and the reaction channel, for example, a synthetic resin tube 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 a metal tube selected from the group consisting of stainless steel, copper and its alloys, and titanium and its alloys can be mentioned. The inner diameters of the supply channel and the reaction channel may be appropriately selected, for example, from sizes usually in the range of 0.1 mm to 1.0 mm, but it is preferably selected from sizes in the range of 0.2 mm to 1.0 mm.

[0072] (E-1-1) and the reaction conditions of the production method of (E-1-2) can also be applied to the following method. <Method 1> General formula [B-0]:

Chemical formula

Chemical formula

Chemical formula

[0073] <Method 2> React the compound of the general formula [B-0-1] with General formula [A-0]:

Chem.

Chem.

[0074] <Method 3> Treat the compound of the general formula [C-0-1] with an oxidizing agent to prepare a compound of general formula [C-0]:

Chem.

[0075] (E-2) Method for producing compound [C-2]

Chem.

[0076] 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 if necessary. Examples of the "base" that can be used in this production method include diisopropylamine, N,N-diisopropylethylamine, triethylamine, N-ethylmorpholine, and 2,6-lutidine.

[0077] When compound [A-2] has a solid-phase carrier in its molecule, that is, when G is a substituent [7] and Z is a solid-phase carrier in compound [A-2], for example, (1) compound [A-1] is filled in an appropriate column and the reaction solution containing compound [B-2] is eluted, or (2) the reaction solution containing compound [A-2] and compound [B-2] is shaken or stirred in a reaction vessel with a filter to carry out this condensation reaction.

[0078] In compound [A-2], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (4) a substituent [7] (however, excluding the case where Z is a solid-phase carrier), for example, (1) compound [A-2] and compound [B-2] are stirred in a reaction solvent in an appropriate reaction vessel, or (2) the solution containing compound [A-2] and the solution containing compound [B-2] are independently supplied through supply channels into a flow reactor or a reaction channel, and the condensation reaction can be carried out by mixing these solutions in the flow reactor or the like. Furthermore, after the condensation reaction, (1) compound [C-2] can be obtained by purifying the reaction mixture using a column, or (2) compound [C-2] can be obtained by adding an appropriate solvent to the reaction mixture, filtering the resulting precipitate, and washing it with an appropriate solvent.

[0079] 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 by nucleoside unit [4b], nucleoside unit [4c], nucleoside unit [4f] or nucleoside unit [4g], it can be produced by the same method as described above.

[0080] (F) Regarding the purification method of compound [C] When compound [C] has a substituent showing very high lipophilicity in its molecule, it can be easily isolated and purified only by crystallization or extraction operations without requiring complicated operations such as column purification. Examples of such compounds include compounds in which, in compound [C-1] and compound [C-2], G is (1) long-chain alkyl-carbonyl, (2) benzoyl substituted with one to five long-chain alkyloxy and / or long-chain alkenyloxy, or (3) substituent [7] (however, excluding the case where Z is a solid-phase carrier). On the other hand, when compound [C] has a solid-phase carrier in its molecule, for example, compound [C] can be purified by packing it into a suitable column and washing compound [C] with a suitable solvent to remove impurities. Examples of such compounds include compounds in which, in compound [C-1] and compound [C-2], G is substituent [7] and Z is a solid-phase carrier. Also, in compound [C-1] and compound [C-2], when G is a silicon substituent, the target compound can be isolated and purified by performing operations such as column purification using a suitable solvent.

[0081] (G) Q within the molecule of compound [C] 1 Elimination 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.

[0082] Hereinafter, the compound [C-1] and the compound [C-2] will be described in detail as examples.

[0083] (G-1) Q within the molecule of compound [C-1] 1 Elimination 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.

[0084] [ka] [In the formula, n, p, B P , G, Q 1 , T, W and X are as defined above.]

[0085] 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]

[0086] In the case where, in compound [C-1], G is substituent [7] and Z is a solid support, for example, this elimination reaction can be carried out by (1) packing compound [C-1] into a suitable column and eluting 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 art, and a single solvent may be used, or two or more solvents may be mixed and used. Examples of the solvent 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; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, and cyclohexane; and halogenated solvents. These solvents may be used in combination. 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, the acidity may be adjusted by combining them with a base (for example, triethylamine) before use. Regarding the amount of the acid used in this elimination reaction, for example, a molar ratio in the range of 1 to 500 times, preferably 2 to 200 times, is appropriate relative to 1 mol of compound [C-1]. The acid that can be used in this elimination reaction is appropriately diluted, for example, to a concentration in the range of 5% to 80% using a suitable solvent, and preferably diluted to a concentration in the range of 5% to 50%. The solvent for dissolving the acid that can be used in this elimination reaction is not particularly limited. For example, chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixed solvent thereof can be mentioned. Also, in this elimination reaction, a scavenger may be used as necessary. Examples of the "scavenger" that can be used in this elimination reaction include ethanol, triisopropylsilane, 1-hydroxybenzotriazole, pyrrole, indole, 2,2,2-trifluoroethanol, methanol, anisole, mercaptoethanol, and thioanisole. The amount of the scavenger used in this elimination reaction is appropriately in the range of 1 to 100 times the molar ratio, preferably in the range of 1 to 50 times the molar ratio, relative to 1 mol of compound [C-1].

[0087] In compound [C-1], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (4) substituent [7] (provided that the case where Z is a solid-phase carrier is excluded), for example, (1) compound [C-1] and an acid are stirred in a suitable reaction solvent in a suitable reaction vessel, or (2) a solution containing compound [C-1] and a solution containing an acid are independently supplied through supply channels into a flow reactor or a reaction channel, and these solutions are mixed in the flow reactor or the like, whereby this elimination reaction can be carried out. 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 art. 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 manufacturing method include aromatic solvents such as benzene, toluene, xylene, and mesitylene; ester solvents such as ethyl acetate and isopropyl acetate; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, and cyclohexane; and halogenated solvents. These solvents may be used in combination. Examples of the "acid" that can be used in this elimination reaction include the same ones as described above. When using these acids, the acidity may be adjusted by combining with a base (e.g., triethylamine) and then used. As for the amount of acid used in this elimination reaction, for 1 mole of compound [C-1], for example, a molar ratio in the range of 1 to 500 times is appropriate, and a range of 2 to 200 times is preferable. The acid that can be used in this elimination reaction is appropriately diluted using a suitable solvent to a concentration in the range of, for example, 5% to 80%, and preferably diluted to a concentration in the range of 5% to 50%. The solvent for dissolving the acid that can be used in this elimination reaction is not particularly limited, and examples include chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixed solvent thereof. Also, in this step, a scavenger may be used as necessary. Examples of the "scavenger" that can be used in this elimination reaction include the same ones as described above. As for the amount of scavenger used in this elimination reaction, for 1 mole of compound [C-1], for example, a molar ratio in the range of 1 to 100 times is appropriate, and a range of 1 to 50 times is preferable. Examples of the means for supplying to the supply flow path that can be used in this elimination reaction include pumps for supplying liquids commonly used in this field, specifically, for example, syringe pumps, plunger pumps, diaphragm pumps, and gear pumps. Examples of the flow reactor that can be used in this elimination reaction include, for example, a microreactor and an in-line mixer such as a static mixer. Examples of the means for guiding from the supply channel to the reaction channel that can be used in this elimination reaction include, for example, a multi-stage collision type micromixer. Examples of the materials of the supply channel and the reaction channel that can be used in this elimination reaction include, for example, a synthetic resin tube 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 a metal tube selected from the group consisting of stainless steel, copper and its alloys, and titanium and its alloys. The inner diameters of the supply channel and the reaction channel that can be used in this elimination reaction may be appropriately selected, for example, from sizes usually in the range of 0.1 mm to 1.0 mm, but it is preferably selected from sizes in the range of 0.2 mm to 1.0 mm.

[0088] As described in the test examples and examples described later, in the method for producing compound [E-1], by using a solvent commonly used in the art as the reaction solvent, a solution containing an acid is added to the reaction mixture containing compound [C-1] produced by subjecting compound [A-1] and compound [B-1] to a condensation reaction, whereby Q 1 The elimination reaction of can be carried out as this continuous reaction in the same system. Also, in the method for producing compound [C-1], by using a solvent commonly used in the art as the reaction solvent, the Q of compound [A-1-1] 1 The elimination reaction of can be carried out, and in the same system, this continuous reaction can be carried out, including subjecting compound [A-1] and compound [B-1] to a condensation reaction 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 art, 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 manufacturing method include aromatic solvents such as benzene, toluene, xylene, and mesitylene; ester solvents such as ethyl acetate and isopropyl acetate; aliphatic solvents such as hexane, pentane, heptane, octane, nonane, and cyclohexane; and halogenated solvents. These solvents may be used in combination.

[0089] Examples of this continuous reaction include in a solvent commonly used in the relevant technical field, Formula [A-1-1]: [Chemical formula] [In the formula, B P is an optionally protected nucleobase; Q 1 is a group that can be eliminated under acidic conditions; W is a lone pair of electrons, an oxygen atom, or a sulfur atom; X is di(C 1-6 alkyl)amino or a substituent represented by general formulas [2-1] to [2-8]: [Chemical formula] [In the formula, * represents the bonding position with P] and is selected from substituents represented by, preferably di(C 1-6 alkyl)amino, more preferably dimethylamino; G is a general formula [7]: [Chemical formula] (In the formula, * represents the bonding position with T; Z is a general formula [8A] to [8D], [8E], [8G], [8H], [8J], [8K], [8N]: [Chemical formula] (In the formula, * represents the bonding position with L; k represents an integer from 0 to 5; R 8a represents a hydrogen atom or C 1-6 alkyl; R 8b each independently represents a long-chain alkyl, either the same or different; R 8c each independently represents the following general formula [9A]:

Chemical formula

[10] :

Chemical formula

[11] :

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0090] As this continuous reaction, for example In a solvent commonly used in the art, Formula [A-1-1]: [Chemical formula] [wherein, Q 1 is trityl, monomethoxytrityl, or dimethoxytrityl, and n, B P , W, X, G, and T are as defined above.] From the compound of trifluoroacetic acid and 2,2,2-trifluoroethanol, optionally in the presence of triisopropylsilane or ethanol, Q 1 can be eliminated. This continuous reaction can be carried out in a flow reactor. For example, a solution containing the compound of the general formula [A-1-1] and a solution containing an acid are fed into a flow reactor to eliminate Q 1 to obtain the compound of the formula [A-1], and a solution containing the compound of the general formula [A-1] and a solution containing the compound of the general formula [B-1] are fed into the next flow reactor to prepare the compound of the general formula [C-1]. Optionally, a flow reactor for feeding a solution containing the compound of the formula [A-1] and a solution containing a scavenger, or a flow reactor for feeding a solution containing an excess of the compound of the formula [B-1] and the compound of the formula [C-1] and a solution containing at least one selected from the group consisting of morpholine, 1-methylpiperazine, and N-ethylmorpholine can be used.

[0091] (G-2) Q within the molecule of compound [C-2] 1 Elimination method

[0092] Since the compound [C-2] is an unstable compound, from the compound [C-2], Q 1 which is substituted at the 5'-position oxygen atom of the 5'-terminal side nucleoside of the compound [C-2]Before detachment, it is preferable to first oxidize the phosphorus atom on the phosphorus bond formed in the condensation reaction from trivalent to pentavalent using an oxidizing agent, and then convert it to a compound represented by the following general formula [D-2] (hereinafter referred to as "compound [D-2]").

[0093]

Chemical formula

[0094] Step 1 Production of compound [D-2] In compound [C-2], when G is 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 the oxidizing agent that can be used in this step include commercially available oxidation solutions for nucleic acid synthesis [oxidation solution - 2, 0.1 mol / L iodine / 78% tetrahydrofuran / 20% pyridine / 2% water, manufactured by Fujifilm Wako Pure Chemical Corporation; oxidation solution, 0.5 M acetone solution of 0.5 M (1S)-(+)-(10-camphorsulfonyl)-oxaziridine, manufactured by Glen Research]. Also, when phosphorylating by oxidizing the phosphorus atom, the oxidation reaction of the phosphorus atom can be carried out according to a method known per se (for example, refer to Current Protocols in Nucleic Acid Chemistry). Examples of the oxidizing agent that can be used in this step include commercially available sulfurization reagents for nucleic acid synthesis [3-{((N,N-dimethylaminomethylene)amino})-3H-1,2,4-dithiazole-5-thione (DDTT), manufactured by Glen Research; 5-phenyl-3H-1,2,4-dithiazol-3-one for nucleic acid synthesis, manufactured by Fujifilm Wako Pure Chemical Corporation]. In this step, it is appropriate to dissolve these oxidizing agents in an appropriate solvent for use.

[0095] In compound [C-2], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) benzoyl substituted with one to five long-chain alkyloxy and / or long-chain alkenyloxy, or (4) substituent [7] (provided that the case where Z is a solid-phase carrier is excluded), the oxidation reaction of the phosphorus atom can be carried out according to a method known per se (see, for example, Nucleic Acids Research, Vol. 21, No. 5, 1213-1217 (1993)). Examples of the oxidizing agent that can be used in this step include (+)-camphorylsulfonyloxaziridine (CSO), (+)-(8,8-dichlorocamphorylsulfonyl)-oxaziridine (DCSO), methyl ethyl ketone peroxide, and tert-butyl hydroperoxide (TBHP).

[0096] Step 2 Production of compound [E-2] By reacting compound [D-2] with an acid, Q 1 substituted at the 5'-position oxygen atom of the nucleoside unit on the 5'-terminal side of compound [D-2] can be eliminated. From compound [D-2], Q 1 in its molecule is eliminated to produce a compound represented by the above general formula [E-2] (hereinafter referred to as "compound [E-2]").

[0097] In compound [D-2], when G is substituent [7] and Z is a solid-phase carrier, for example, it can be carried out by packing compound [D-2] in a suitable column and eluting a solution containing an acid, or by shaking or stirring a solution containing compound [D-2] and an acid in a reaction vessel with a filter. Q in the molecule of compound [D-2] 1 The elimination reaction can be carried out according to a method known per se (see, for example, Current Protocols in Nucleic Acid Chemistry). Examples of the acid that can be used in this step include commercially available deblocking solutions for nucleic acid synthesis [for example, Deblocking Solution-1, 3 w / v% trichloroacetic acid / dichloromethane solution (manufactured by Fujifilm Wako Pure Chemical Corporation), Deblocking Mix 3% dichloroacetic acid / dichloromethane solution (manufactured by Glen Research Corporation)].

[0098] In compound [D-2], when G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (4) substituent [7] (however, excluding the case where Z is a solid-phase carrier), for example, (1) compound [D-2] and an acid are stirred in a suitable reaction solvent in a suitable reaction vessel, or (2) a solution containing compound [D-2] and a solution containing an acid are independently supplied into a flow reactor or a reaction channel through supply channels, and the solutions are mixed in the flow reactor or the like. Q within the molecule of compound [D-2] 1 The elimination reaction can be carried out according to a method known per se (for example, see Nucleic Acids Research, Vol. 21, No. 5, 1213-1217 (1993)). Examples of the acid that can be used in this step include dichloroacetic acid and trichloroacetic acid.

[0099] (H) Regarding the final deprotection and isolation step of nucleic acid compounds When compound [C-1], compound [D-2], compound [E-1] or compound [E-2] has a protecting group in its molecule, a compound in which all the protecting groups are removed can be produced by performing a deprotection treatment according to the type or nature of the protecting group. For example, according to the deprotection methods described in "Green’s PROTECTIVE GROUPS in ORGANIC SYNTHESIS, 4th Edition, 2006", all the protecting groups of the compound can be removed. Specifically, for example, by treating with (1) aqueous ammonia, (2) aqueous ammonia / ethanol or (3) a mixed solution of aqueous ammonia and aqueous methylamine solution, the substituent [6] in the molecule of compound [C-1], compound [D-2], compound [E-1] or compound [E-2] and the protecting group of the amino group or hydroxyl group of the nucleobase can be removed. Also, for example, the same "acid" as described in the "Method for Removing Q 1 in the molecule of compound [C-1]", the same "acid" as described in the "Step 2: Production of compound [E-2]" in the "Method for Removing Q 1 in the molecule of compound [C-2]", or by treating with a solution obtained by diluting hydrochloric acid or acetic acid with an appropriate solvent, the protecting group of the amino group at the 3'-position of the nucleoside on the 3'-terminal side of compound [C-1] and the group that can be removed under acidic conditions substituted for the hydroxyl group at the 5'-position of the nucleoside on the 5'-terminal side of compound [D-2] can be removed. When removing the group that can be removed under acidic conditions substituted for the hydroxyl group at the 5'-position of the nucleoside on the 5'-terminal side of compound [D-2] after removing the protecting group of the nucleobase moiety, a diluted acid with water is used. When the nucleobase moiety is protected, a diluted acid with an appropriate organic solvent is used.

[0100] (I) Regarding the purification and separation step The compound in which all the protecting groups of compound [C-1] or compound [E-1] are removed can be separated and purified from the reaction mixture by ordinary separation and purification means, for example, extraction, concentration, neutralization, filtration, centrifugation, recrystallization, from C8 to C 18The 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.

[0101] 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 a conventional separation and purification method, for example, extraction, concentration, neutralization, filtration, centrifugation, recrystallization, or separation of 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.

[0102] (J) Production of compound [A] Compound [A] is produced, for example, by introducing substituent [6] into the hydroxyl group of the compound corresponding to compound [A] according to a known method.

[0103] The following presents representative examples to illustrate the production method of compound [A].

[0104] (J-1) Production of compound [A-1] Compound [A] composed of one to a plurality of nucleoside units [4d] with a phosphorous bond [5] between each nucleoside unit can be produced, for example, according to the methods described in the following (i) to (iv).

[0105] (i) Production of compound [A-1] where G is a silicon substituent and T is a single bond

Chemical formula

[0106] Step 1 Production of the compound represented by the above general formula [A-1a-Q1] (hereinafter referred to as "compound [A-1a-Q1]") Using the compound represented by the above general formula

[21] (hereinafter referred to as "compound

[21] ") and the compound represented by the above general formula [20A] (hereinafter referred to as "compound [20A]"), compound [A-1a-Q1] can be produced by introducing a silicon substituent into the hydroxyl group at the 5'-end of compound

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

[0107] Process 2: Production of Compound [A-1a] Compound [A-1a] can be produced by subjecting Compound [A-1a-Q1] to an acid treatment. The "acid" that can be used in this process can be the same as the "acid" described in the "Method for the elimination of Q in Compound [C-1]". 1 in the molecule. As the amount of the acid that can be used in this process, for 1 mol of Compound [A-1a-Q1], for example, a molar ratio in the range of 1 to 500 times is appropriate, and a range of 2 to 200 times is preferable. The acid that can be used in this process may be diluted with a suitable solvent and is not particularly limited. For example, chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixed solvent thereof can be mentioned.

[0108] Also, in this process, a scavenger may be used as necessary. The "scavenger" that can be used in this process can be the same as the "scavenger" described in the "Method for the elimination of Q in Compound [C-1]". 1 in the molecule. As the amount of the scavenger that can be used in this process, for 1 mol of Compound [A-1a-Q1], for example, a molar ratio in the range of 1 to 100 times is appropriate, and a range of 1 to 50 times is preferable.

[0109] (ii) Production of Compound [A-1] where 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

Chemical formula

[0110] The compound represented by the above general formula [A-1b] (hereinafter referred to as "compound [A-1b]") is a compound [A-1] in which G is (1) a long-chain alkyl-carbonyl, (2) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy, or (3) a substituent [7], and T is a single bond. An example of a method for producing compound [A-1b] is described below.

[0111] Step 1 Production 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 the 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 compound [20B] in which Y is a hydroxyl group is used in this step, it can be carried out in the range of -20°C to 100°C using a condensing agent in the presence or absence of a base. When compound [20B] in which Y is a halogen is used in this step, it can be carried out in the range of -20°C to 100°C in the presence of a base. Examples of the condensing agent that can be used in this step include 1,1'-oxalyldiimidazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, dicyclohexylcarbodiimide, diethyl cyanophosphonate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate. Examples of the base that can be used in this step include organic bases such as triethylamine, N,N-diisopropylethylamine, N,N-dimethylaniline, pyridine, and 1,8-diazabicyclo[5.4.0]-7-undecene. The solvent that can be used in this step is not particularly limited. 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; or a mixed solvent 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 the additive that can be used in this step 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 suitably within the range of 10 minutes to 24 hours. The usage amounts of the compound

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

[0112] Process 2: Production of Compound [A-1b] Compound [A-1b] can be produced by subjecting Compound [A-1b-Q1] to acid treatment.

[0113] The "acid" that can be used in this process can be the same as the "acid" described in the "Method for Elimination of Q in Compound [C-1]". 1 in the molecule of Compound [C-1]. As the amount of acid used in this process, for 1 mol of Compound [A-1b-Q1], for example, a molar ratio in the range of 1 to 500 times is appropriate, and a range of 2 to 200 times is preferred. The acid that can be used in this process may be diluted with a suitable solvent and is not particularly limited. For example, chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixed solvent thereof can be mentioned.

[0114] Also, in this process, a scavenger may be used as necessary. The "scavenger" that can be used in this process can be the same as the "scavenger" described in the "Method for Elimination of Q in Compound [C-1]". 1 in the molecule of Compound [C-1]. As the amount of scavenger used in this process, for 1 mol of Compound [A-1b-Q1], for example, a molar ratio in the range of 1 to 100 times is appropriate, and a range of 1 to 50 times is preferred.

[0115] (iii) Production of Compound [A-1] where 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]

Chemical formula

[0116] The compound represented by the above general formula [A-1c] (hereinafter referred to as "compound [A-1c]") is a compound [A-1] in which G is (1) 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 the method for producing compound [A-1c] is described below.

[0117] Step 1 Production of the compound represented by the above general formula

[23] (hereinafter referred to as "compound

[23] ") Compound

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

[22] (hereinafter referred to as "compound

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

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

[0118] Step 2 Production of the compound represented by the above general formula

[24] (hereinafter referred to as "compound

[24] ") Compound

[24] can be produced by eliminating the trityl group in the molecule of compound

[23] with an acid.

[0119] Process 3 Production of the compound represented by the general formula [A-1c-Q1] (hereinafter referred to as "compound [A-1c-Q1]"). Compound [A-1c-Q1] can be produced by condensing compound

[24] and the compound represented by the general formula

[25] (hereinafter referred to as "compound

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

[0120] Process 4 Production of compound [A-1c]. Compound [A-1c] can be produced by subjecting compound [A-1c-Q1] to acid treatment. The "acid" that can be used in this process can be the same as the "acid" described in the "method for eliminating Q in the molecule of compound [C-1]". 1 The same ones as the "acid" described in the "method for eliminating Q in the molecule of compound [C-1]" can be mentioned. As the amount of the acid that can be used in this process, for example, in the range of 1-fold to 500-fold in molar ratio with respect to 1 mol of compound [A-1c-Q1] is appropriate, and in the range of 2-fold to 200-fold is preferable. The acid that can be used in this process may be diluted with an appropriate solvent and is not particularly limited. For example, chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol or a mixed solvent thereof can be mentioned.

[0121] Also, in this process, a scavenger may be used as necessary. The "scavenger" that can be used in this process can be the same as the "scavenger" described in the "method for eliminating Q in the molecule of compound [C-1]". 1 The same ones as the "scavenger" described in the "method for eliminating Q in the molecule of compound [C-1]" can be mentioned. As the amount of the scavenger that can be used in this process, for example, in the range of 1-fold to 100-fold in molar ratio with respect to 1 mol of compound [A-1c-Q1] is appropriate, and in the range of 1-fold to 50-fold is preferable.

[0122] (iv) Production of compound [A-1] wherein G is substituent [7] and T is a single bond

Chemical formula

[0123] The compound represented by the above general formula [A-1d] (hereinafter referred to as "compound [A-1d]") is a compound [A-1] wherein G is substituent [7] and T is a single bond. An example of the production method of compound [A-1d] is described below.

[0124] Step 1 Production of the compound represented by the above general formula [A-1d-Q1] (hereinafter referred to as "compound [A-1d-Q1]") By condensing the compound represented by the above general formula [20D] (hereinafter referred to as "compound [20D]") with the compound represented by the above general formula

[21] (hereinafter referred to as compound

[21] ), compound [A-1d-Q1] can be produced. The condensation reaction can be carried out according to a method known per se. Compound [20D] is in the carboxylic acid form, but its reactive derivative can also be used in this step. Examples of the reactive derivative of compound [20D] include those commonly used in ester condensation formation reactions, such as acid halides (e.g., acid chlorides, acid bromides). When using compound [20D], the reaction can be carried out in the range of -20°C to 100°C using a condensing agent in the presence or absence of a base. Examples of the condensing agent that can be used in this step include 1,1'-oxalyldiimidazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, dicyclohexylcarbodiimide, diethyl cyanophosphonate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate. Examples of the base that can be used in this step include organic bases such as triethylamine, N,N-diisopropylethylamine, N,N-dimethylaniline, pyridine, and 1,8-diazabicyclo[5,4,0]-7-undecene. The solvent that can be used in this step is not particularly limited, and 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; or a mixed solvent thereof. Further, additives can be used as necessary. Examples of the additive that can be used in this step include 4-dimethylaminopyridine, 1-hydroxybenzotriazole, and 1-hydroxy-7-azabenzotriazole. The reaction time varies depending on the types of raw materials used, reaction temperature, etc., but usually, a range of 10 minutes to 24 hours is appropriate. As for the usage amounts of compound

[21] and the condensing agent, for example, a range of 1 to 1.5 times moles relative to 1 mole of compound [20D] is appropriate. As for the usage amount of the base, for example, it is in the range of 1 equivalent to 10 equivalents, preferably in the range of 1 equivalent to 4 equivalents, relative to compound [20D].

[0125] Step 2 Production of compound [A-1d] Compound [A-1d-Q1] can be produced by subjecting it to acid treatment to obtain compound [A-1d]. The "acid" that can be used in this step can be the same as the "acid" described in the "method for eliminating Q in the molecule of compound [C-1]". 1 The "acid" that can be used in this step can be the same as the "acid" described in the "method for eliminating Q in the molecule of compound [C-1]". As the amount of the acid used in this step, for 1 mole of compound [A-1d-Q1], for example, a molar ratio in the range of 1-fold to 500-fold is appropriate, and a range of 2-fold to 200-fold is preferred. The acid that can be used in this step may be diluted with a suitable solvent and is not particularly limited. For example, chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixed solvent thereof can be mentioned. Also, in this step, a scavenger may be used as necessary. The "scavenger" that can be used in this step can be the same as the "scavenger" described in the "method for eliminating Q in the molecule of compound [C-1]". 1 The "scavenger" that can be used in this step can be the same as the "scavenger" described in the "method for eliminating Q in the molecule of compound [C-1]". As the amount of the scavenger used in this step, for 1 mole of compound [A-1d-Q1], for example, a molar ratio in the range of 1-fold to 100-fold is appropriate, and a range of 1-fold to 50-fold is preferred.

[0126] For example, compound [20D] can be produced according to the production method described below.

Chemical formula

[0127] Step 1 Production of the compound represented by the above general formula

[28] (hereinafter referred to as “compound

[28] ”). The compound represented by the above general formula

[26] (hereinafter referred to as "compound

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

[27] (hereinafter referred to as "compound

[27] ") to produce compound

[28] . The condensation reaction can be carried out according to a method known per se. As the reagents, reaction conditions, etc. that can be used in this step, those similar to those in the above "Production of compound [A-1b-Q1]" can be used.

[0128] Step 2 Production of compound [20D] Compound [20D] can be produced by subjecting compound

[28] to ester hydrolysis. The ester hydrolysis reaction can be carried out according to a method known per se. The solvent that can be used in this step is not particularly limited, and 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, or a mixed solvent thereof. This step is carried out in the presence of a base such as sodium hydroxide, potassium hydroxide or lithium hydroxide within the range of 20 °C to 100 °C. The reaction time varies depending on the type of raw materials used, the reaction temperature, etc., but usually, a range of 10 minutes to 24 hours is appropriate.

[0129] In addition, compound

[26] can be produced, for example, according to the methods described in the following (a) to (j). (a) For example, by using a primary amine compound available as a commercial product or by aminating an alkyl halide available as a commercial product, Z is a substituent [8A], and R 8a is a hydrogen atom, and R 8b is a long-chain alkyl, compound

[26] can be produced. (b) For example, by alkylating a primary amine compound available as a commercial product, Z is a substituent [8A] or a substituent [8B], and R8a is C 1-6 is alkyl, and R 8b are each the same or different and are long-chain alkyls, the compound

[26] can be produced. The alkylation reaction can be carried out according to a method known per se. (c) According to a known method (see, for example, Cancer Res., 2008 Nov 1; 68(21):8843 - 8851, Chem.Sci., 2016, 7, 2308 - 2321), the compound

[26] in which Z is a substituent [8C] can be produced. (d) For example, methyl phthalate is condensed with 1-(tert-butoxycarbonyl)piperazine, then the ester moiety is hydrolyzed using an alkali such as sodium hydroxide, and further condensed with the compound

[26] in which Z is a substituent [8A], and then the tert-butoxycarbonyl group is eliminated using an acid such as trifluoroacetic acid, whereby the compound

[26] in which Z is a substituent [8D] can be produced. 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 according to a method known per se. (e) For example, by alkylating one hydroxyl group of ethane-1,2-diol using an alkyl halide, a compound

[26] in which Z is a substituent [8E] and R 8e is a long-chain alkyl group can be produced. For example, by long-chain alkyl-carbonylating one hydroxyl group of ethane-1,2-diol, a compound

[26] in which Z is a substituent [8E] and R 8e is long-chain alkyl-carbonyl can be produced. As the compound used for long-chain alkyl-carbonylation, for example, the corresponding carboxylic acid compound or its reactive derivative can be used. Examples of the reactive derivative include those usually used in an ester condensation formation reaction, such as acid halides (e.g., acid chloride, acid bromide). For example, by condensing one hydroxyl group of ethane-1,2-diol with the compound [20C], a compound

[26] in which Z is a substituent [8E] and R8e Compound

[26] in which the benzoyl group is substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy groups can be produced. (f) For example, by using 2-amino-ethanol instead of ethane-1,2-diol and following the same method as the production method of the compound (18) in which Z is the substituent [8E], the compound

[26] in which Z is the substituent [8F] can be produced. (g) For example, after condensing 9-fluorenylmethyloxycarbonyl-phenylalanine with the compound

[26] in which Z is the substituent [8A], and then eliminating the 9-fluorenylmethyloxycarbonyl group with piperidine, the compound

[26] in which Z is the substituent [8G] can be produced. The condensation reaction and the deprotection reaction of the 9-fluorenylmethyloxycarbonyl group can be carried out according to methods known per se. (h) For example, after producing according to the same method as the production method of the compound

[26] in which Z is the substituent [8E] by using 1-(tert-butoxycarbonyl)piperazine instead of ethane-1,2-diol, and then deprotecting the tert-butoxycarbonyl group in the molecule with an acid, the compound

[26] in which Z is the substituent [8H] can be produced. (i) According to known methods (for example, refer to Japanese Patent No. 5705512, Tetrahedron Letters, Vol. 53, 1936 - 1939 (2012), International Publication No. 2014 / 189142A1, International Publication No. 2016 / 060135A1, International Publication No. 2016 / 140232A1), the compound

[26] in which Z is the substituent [8I], the substituent [8J], the substituent [8K], the substituent [8L], the substituent [8N] can be produced. (j) For example, 9H-xanthen-9-one having a hydroxyl group is treated with a base such as sodium hydride, and an appropriate halogenated long-chain alkyl is allowed to act thereon, whereby 9H-xanthen-9-one having a corresponding long-chain alkyloxy can be produced. Further, by allowing phenylmagnesium bromide which may be further substituted to act, a compound

[26] in which Z is a substituent [8M] can be produced. In addition, a desired compound

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

[0130] Also, according to a known method (see, for example, International Publication No. 91 / 09033A1), a compound

[21] where n = 1 can be produced, and then, according to the method described below, a compound

[21] where n>1 can be produced. [Chemical formula] [In the formula, n, B P , D, Q 1 , X and W have the same meanings as described above, Ac represents acetyl.]

[0131] Process 1: Production of the compound represented by the general formula

[30] (hereinafter referred to as "compound

[30] ") In the presence of a base, the compound represented by the above general formula

[29] (see, for example, International Publication No. 91 / 09033A1) is acetylated with acetic anhydride to produce a compound

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

[0132] Process 2: Production of the compound represented by the general formula

[31] (hereinafter referred to as "compound

[31] ") By subjecting the compound

[30] to an acid treatment, a compound

[31] can be produced. The elimination reaction of Q 1 can be carried out according to a method known per se.

[0133] Process 3: Production of the compound represented by the general formula

[33] (hereinafter referred to as "compound

[33] ") Compound

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

[32] (hereinafter referred to as "compound

[32] ") to produce compound

[33] . The condensation reaction can be carried out according to a method known per se (for example, refer to International Publication No. 91 / 09033A1). Note that compound

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

[0134] Process 4: Production of compound

[21] For example, compound

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

[33] using an alkali metal alkoxide such as sodium methoxide. The reaction for removing acetyl can be carried out according to a method known per se (for example, refer to Tetrahedron Letters, Vol. 50, 1751 - 1753 (2009)).

[0135] (J-2) Production of compound [A-2] A compound [A] composed of one to a plurality of nucleoside units selected from the group consisting of nucleoside unit [4a], nucleoside unit [4b], and nucleoside unit [4c], and having a phosphorous bond [5] between each nucleoside unit can be produced, for example, according to the methods described in the following (i) to (iv).

[0136] (i) Production of compound [A-2] where G is a silicon substituent and T is a single bond

Chemical formula

[0137] Process 1: Production of the compound represented by the general formula [A-2a-Q1] (hereinafter referred to as "compound [A-2a-Q1]") By using compound [20A] for the compound represented by the general formula

[34] (hereinafter referred to as "compound

[34] ") and introducing a silicon substituent to the hydroxyl group at the 3'-position of the nucleoside unit on the 3'-terminal side of compound

[34] , compound [A-2a-Q1] can be produced. The introduction reaction of this silicon substituent can be carried out according to a method known per se.

[0138] Process 2: Production of compound [A-2a] Compound [A-2a] can be produced by subjecting compound [A-2a-Q1] to acid treatment. The "acid" that can be used in this step can be the same as the "acid" described in the "elimination reaction of Q 1 in the molecule of compound [D-2]". As the amount of the acid that can be used in this step, for 1 mol of compound [A-2a-Q1], for example, a range of 1 to 500 times the molar ratio is appropriate, and a range of 2 to 200 times the molar ratio is preferred. The acid that can be used in this step may be diluted with a suitable solvent and is not particularly limited. For example, chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixed solvent thereof can be mentioned.

[0139] Also, in this step, a scavenger may be used as necessary. The "scavenger" that can be used in this step can be the same as the "scavenger" described in the "elimination reaction of Q 1 in the molecule of compound [D-2]". As the amount of the scavenger that can be used in this step, for 1 mol of compound [A-2a-Q1], for example, a range of 1 to 100 times the molar ratio is appropriate, and a range of 1 to 50 times the molar ratio is preferred.

[0140] (ii) Production of compound [A-2] where 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 [Chemical formula] [In the formula, n, B P , G 2 , Q 1 , R 4a , X, Y and W are as defined above.]

[0141] The compound represented by the general formula [A-2b] (hereinafter referred to as "compound [A-2b]") is a compound [A-2] in which G is (1) a long-chain alkyl-carbonyl, (2) 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.

[0142] An example of the method for producing the compound [A-2b] is described below.

[0143] Process 1: Production of the compound represented by the 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 according to a method known per se. When using compound [20B] in which Y is a hydroxyl group in this step, it can be carried out in the range of -20°C to 100°C using a condensing agent in the presence or absence of a base. When using compound [20B] in which Y is a halogen in this step, it can be carried out in the range of -20°C to 100°C in the presence of a base. Examples of the condensing agent that can be used in this process include 1,1'-oxalyldiimidazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, dicyclohexylcarbodiimide, diethyl cyanophosphonate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, and 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate. Examples of the base that can be used in this process 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 process is not particularly limited, and 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; or a mixed solvent thereof. In addition, when using the compound [20B] in which Y is a hydroxyl group in this process, an additive can be used as needed. Examples of the additive that can be used include 4-dimethylaminopyridine, 1-hydroxybenzotriazole, and 1-hydroxy-7-azabenzotriazole. The reaction time varies depending on the type of raw materials used, the reaction temperature, etc., but usually, a range of 10 minutes to 24 hours is appropriate. As for the usage amounts of the compound [20B] and the condensing agent, for example, a range of 1 to 1.5 moles per mole of the compound

[34] is appropriate. As for the usage amount of the base, for example, it is in the range of 1 equivalent to 10 equivalents, preferably in the range of 1 equivalent to 4 equivalents, relative to the compound

[34] .

[0144] Process 2: Production of compound [A-2b] Compound [A-2b-Q1] can be converted to compound [A-2b] by acid treatment.

[0145] }The "acid" that can be used in this step is the same as the "acid" described in the "elimination reaction of Q in the molecule of compound [D-2]". 1 can be mentioned. As the amount of the acid that can be used in this step, for 1 mole of compound [A-2b-Q1], for example, a molar ratio in the range of 1 to 500 times is appropriate, and a range of {2 to 200 times is preferred. The acid that can be used in this step may be diluted with a suitable solvent and is not particularly limited. For example, chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixed solvent thereof can be mentioned.

[0146] In addition, in this step, a scavenger may be used as necessary. The "scavenger" that can be used in this step is the same as the "scavenger" described in the "elimination reaction of Q in the molecule of compound [D-2]". 1 can be mentioned. As the amount of the scavenger that can be used in this step, for 1 mole of compound [A-2b-Q1], for example, a molar ratio in the range of 1 to 100 times is appropriate, and a range of 1 to 50 times is preferred. [[ID={23]] "

[0147] <{ (iii) Production of compound [A-2] where 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]

Chemical Formula

[0148] The compound represented by the general formula [A-2c] (hereinafter referred to as "compound [A-2c]") is a 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] .

[0149] An example of the production method of compound [A-2c] is described below.

[0150] Process 1: Production of the compound represented by the general formula

[36] (hereinafter referred to as "compound

[36] ") Compound

[36] can be produced by condensing compound

[24] with the compound represented by the general formula

[35] (hereinafter referred to as "compound

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

[0151] Process 2: Production of the compound represented by the general formula [A-2c-Q1] (hereinafter referred to as "compound [A-2c-Q1]") Compound [A-2c-Q1] can be produced by reacting an oxidizing agent with compound

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

[36] , more preferably 10 to 50 times the molar amount.

[0152] Process 3: Production of compound [A-2c] Compound [A-2c-Q1] can be produced by subjecting it to acid treatment to obtain compound [A-2c]. The "acid" that can be used in this step can be the same as the "acid" described in the "elimination reaction of Q within the molecule of compound [D-2]". 1 Specific examples can be given. As the amount of the acid used in this step, for 1 mole of compound [A-2c-Q1], for example, a molar ratio in the range of 1-fold to 500-fold is appropriate, and a range of 2-fold to 200-fold is preferred. The acid that can be used in this step may be diluted with a suitable solvent, and is not particularly limited. For example, chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixed solvent thereof can be mentioned. Also, in this step, a scavenger may be used as needed. The "scavenger" that can be used in this step can be the same as the "scavenger" described in the "elimination reaction of Q within the molecule of compound [D-2]". 1 Specific examples can be given. As the amount of the scavenger used in this step, for 1 mole of compound [A-2c-Q1], for example, a molar ratio in the range of 1-fold to 100-fold is appropriate, and a range of 1-fold to 50-fold is preferred.

[0153] (iv) Production of compound [A-2] where G is substituent [7] and T is a single bond

Chemical formula

[0154] An example of a method for producing compound [A-2d] is described below.

[0155] Process 1 By condensing compound [20D] with compound

[34] , a compound represented by the above general formula [A-2d-Q1] (hereinafter referred to as "compound [A-2d-Q1]") can be produced. The condensation reaction can be carried out according to a method known per se. Although compound [20D] is a carboxylic acid form, its reactive derivative can also be used in this step. Examples of the reactive derivative of compound [20D] include those commonly used in ester condensation formation reactions, such as acid halides (e.g., acid chlorides, acid bromides). When using compound [20D], the reaction can be carried out within the range of -20°C to 100°C using a condensing agent in the presence or absence of a base. Examples of the condensing agent that can be used in this step include 1,1'-oxalyldiimidazole, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, dicyclohexylcarbodiimide, diethyl cyanophosphonate, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, 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, 1,8-diazabicyclo[5,4,0]-7-undecene. The solvents that can be used in this process are not particularly limited. For example, 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, or mixed solvents thereof can be mentioned. Further, additives can be used as necessary. Examples of the additives that can be used in this process include 4-dimethylaminopyridine, 1-hydroxybenzotriazole, and 1-hydroxy-7-azabenzotriazole. The reaction time varies depending on the types of raw materials used, reaction temperature, etc., but usually, a range of 10 minutes to 24 hours is appropriate. As the usage amounts of compound [20D] and the condensing agent, for example, a range of 1 to 1.5 moles per mole of compound

[34] is appropriate. As the usage amount of the base, for example, it is in the range of 1 equivalent to 10 equivalents, preferably in the range of 1 equivalent to 4 equivalents, relative to compound

[34] .

[0156] Process 2 Compound [A-2d] can be produced by subjecting compound [A-2d-Q1] to acid treatment. The "acid" that can be used in this process can be the same as the "acid" described in the "elimination reaction of Q 1 in the intramolecular of compound [D-2]". As the usage amount of the acid that can be used in this process, for example, a range of 1 to 500 times the amount in molar ratio, preferably a range of 2 to 200 times the amount, per mole of compound [A-2d-Q1] is appropriate. The acid that can be used in this process may be diluted with a suitable solvent and is not particularly limited. For example, chloroform, dichloromethane, 1,1-dichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 1,2-dichloroethylene, 2,2,2-trifluoroethanol, or a mixed solvent thereof can be mentioned. Also, in this step, a scavenger may be used as necessary. The "scavenger" that can be used in this step is the same as the "scavenger" described in the "elimination reaction of Q in the molecule of the compound [D-2]". 1 The same ones as the "scavenger" described in the "elimination reaction of Q in the molecule of the compound [D-2]" can be mentioned. As the amount of the scavenger that can be used in this step, for 1 mol of the compound [A-2d-Q1], for example, a range of 1-fold to 100-fold in molar ratio is appropriate, and a range of 1-fold to 50-fold is preferable.

[0157] Also, according to a known method (Current Protocols in Nucleic Acid Chemistry), the compound

[34] with n = 1 can be produced, and according to a known method (for example, refer to U.S. Patent Application Publication No. 2010 / 273999A1), the compound

[34] with n > 1 can be produced.

[0158] The compound "A-2" is a compound in which each nucleoside unit constituting the compound is the nucleoside unit [4a], but a compound in which all or part of the nucleoside units [4a] are replaced by the nucleoside unit [4b] or the nucleoside unit [4c] can also be produced by using the same method as above.

[0159] (K) Production of compound [B] The compound [B] is produced, for example, by introducing a substituent [1] into the hydroxyl group of the compound corresponding to the compound [B] according to a known method.

[0160] Hereinafter, representative examples will be introduced to explain the production method of the compound [B].

[0161] (K-1) Production of compound [B-1] A compound [B] composed of one to a plurality of nucleoside units [4h] and having a phosphorous bond [5] between each nucleoside unit can be produced, for example, according to the method described below. [Chemistry] [In the formula, p, B P , Q 1 , D, X and W have the same meanings as described above.]

[0162] Process 1: Production of the compound represented by the general formula

[39] (hereinafter referred to as "compound

[39] ") Compound

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

[37] (hereinafter referred to as "compound

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

[38] (hereinafter referred to as "compound

[38] "). The condensation reaction can be carried out according to a method known per se (see, for example, US Patent Application Publication No. 2014 / 0330006A1, International Publication No. 2012 / 043730A1, International Publication No. 2013 / 082548A1).

[0163] Process 2: Production of compound [B-1] Compound [B-1] can be produced by condensing the compound represented by the above general formula

[40] (hereinafter referred to as "compound

[40] ") with compound

[39] . The condensation reaction can be carried out according to a method known per se (see, for example, US Patent Application Publication No. 2014 / 0330006A1, International Publication No. 2012 / 043730A1, International Publication No. 2013 / 082548A1, International Publication No. 91 / 09033A1). Compound

[40] can be produced by using a method similar to the method for producing the above compound

[21] .

[0164] (K-2) Production of compound [B-2] Compound [B] composed of one or more nucleoside units selected from the group consisting of nucleoside unit [4e], nucleoside unit [4f] and nucleoside unit [4g], and having a phosphorous bond [5] between each nucleoside unit can be produced, for example, according to the method described below. [Chemistry] [In the formula, p, B P , Hal, Q 1 , D, R 4a , X and W are as defined above.]

[0165] Process 1: Production of the compound represented by the general formula

[42] (hereinafter referred to as "compound

[42] ") Compound

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

[41] (hereinafter referred to as "compound

[41] ") to produce compound

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

[0166] Process 2: Production of compound [B-2] According to a known method, a compound represented by the above general formula

[43] (hereinafter referred to as "compound

[43] ") is allowed to act on compound

[42] , and a substituent containing a phosphorus atom is introduced into the hydroxyl group at the 3'-position of the nucleoside unit on the 3'-terminal side, whereby compound "B-2" can be obtained. In this step, an activator can be used if necessary. The solvent used in this step is not particularly limited, and examples thereof include acetonitrile and tetrahydrofuran. The amount of compound

[42] used is preferably 1 to 20 molar amounts, more preferably 1 to 10 molar amounts, relative to 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, 2,4,6-collidine / N-methylimidazole. The amount of the "activator" used is appropriately 1 to 20 times the molar amount, preferably 1 to 10 times the molar amount, relative to compound

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

[0167] Compound [B-2] is a compound in which each nucleoside unit constituting the compound is nucleoside unit [4e], but a compound in which all or part of nucleoside unit [4e] is replaced by nucleoside unit [4f] or nucleoside unit [4g] can also be produced according to the same method as above.

Examples

[0168] The present invention will be described in more detail below with reference to Examples, Comparative Examples, and Test Examples, but the present invention is not limited thereto. Note that the "conversion yield (%)" means the ratio of the raw material converted into the target product, and is calculated as "{peak area (%) corresponding to the target product detected by high performance liquid chromatography (hereinafter referred to as "HPLC")} ÷ {peak area (%) corresponding to the raw material detected by HPLC + peak area (%) corresponding to the target product detected by HPLC} × 100". HPLC conditions: 0.5 mg of the product is dissolved in acetonitrile or 20% water / acetonitrile, and HPLC analysis is performed under the following conditions. The coupling efficiency is calculated by using the integrated value of the peak area obtained by absorption at UV = 264 nm in HPLC. <ODS conditions> Column: Waters XBridge C18 (2.5 μm, 4.6 × 50 mm), 60°C Detection wavelength: 264 nm Mobile phase A: 20 mM AcONH4aq. Mobile phase B: MeCN Flow rate: 0.75 mL / min Gradient: 40 - 95% B (0 - 15 min), 95% B (15 - 24 min), 40% B (24 - 30 min) LC / MS Conditions: Equipment Used: Ultra-High Performance Liquid Chromatograph ACQUITY UPLC (waters) Quadrupole Time-of-Flight Mass Spectrometer SYNAPT-MS (waters) Column: ACQUITY UPLC BEH C18 1.7 μm, 2.1×50 mm (waters) Temperature: 50 °C Flow Rate: 0.2 mL / min Mobile Phase: 10 mM Ammonia Water Mobile Phase: MeCN Gradient: 50 - 95% B (4 min) Detector 1: UV264 nm Detector 2: Quadrupole Time-of-Flight Mass Spectrometer Ionization Method: ESI+ Measurement Range: 100 - 2000 m / z

[0169] Example 1 4-(Octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl (hereinafter referred to as "G1-suc-morT-OFF")

[0170] Production 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 the mixture was stirred at room temperature for 7 hours. The mixture was concentrated under reduced pressure, 150 mL of acetone was added to the residue, and the mixture was stirred for 16 hours. The precipitate was suction filtered, washed with acetone (400 mL), and dried under reduced pressure at 30 °C for 3 hours to obtain G1-suc (29.1 g, 96.6%) as a white powder.

[0171] Production of 4-(octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl (hereinafter referred to as "G1-suc-morT-OFF"). 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. Then, 1-((2R,6S)-6-(hydroxymethyl)-4-tritylmorpholin-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (hereinafter referred to as "morT-OH") (18 g, 1.0 eq.) and 4-(N,N-dimethylamino)pyridine 4.57 g were added, and the mixture was stirred on a 70 °C water bath for 1 hour. After cooling to room temperature, 0.1 M aqueous sodium dihydrogen phosphate solution was added and stirred briefly. Then, the aqueous layer was removed, and the organic layer was washed once with 0.1 M aqueous sodium dihydrogen phosphate solution and saturated brine diluted twice with water. The aqueous layers were combined and extracted with dichloromethane. The combined organic layers were dried over anhydrous sodium sulfate. After filtration, the solvent was distilled off, and dried under reduced pressure to obtain 4-(octadecylamino)-4-oxobutanoic acid [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methyl (hereinafter referred to as "G1-suc-morT-ON") (white amorphous, 28.1 g, 89.5%). This was dissolved in 140 mL of dichloromethane and stirred on an ice bath. While stirring, 20 mL of 2,2,2-trifluoroethanol and 10.3 mL of triisopropylsilane were added and stirred briefly, and then 5.1 mL of trifluoroacetic acid was added dropwise. One hour after the addition was completed, ice was added to 100 mL of saturated aqueous sodium bicarbonate solution, and the reaction solution was poured into the cooled solution while stirring. After confirming that the aqueous layer had a pH of 7-8, the aqueous layer was extracted with dichloromethane. The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated. Column chromatography purification was performed on silica gel using a dichloromethane-methanol mixed solution as the mobile phase, and dried under reduced pressure to obtain 19.89 g of G1-suc-morT-OFF as a powder.

[0172] 11H-NMR (CDCl3): δ 8.90 (1H, bs); 7.25 (1H, d, J = 1.6 Hz); 5.72 (1H, dd, J = 9.6 Hz, 2.6 Hz); 5.65 (1H, m); 4.14 (2H, d, J = 5.2 Hz); 3.98 (1H, m); 3.23 (2H, dd, J = 12.8 Hz, 7.0 Hz); 3.12 (2H, dd, J = 12 Hz, 2.6 Hz); 2.95 (2H, dd, J = 12.8 Hz, 1.8 Hz); 2.60 - 2.75 (4H, m); 2.47 (2H, t, J = 6.8 Hz); 1.95 (3H, d, J = 1.6 Hz); 1.48 (2H, m), 1.21 - 1.34 (29H, m); 0.88 (3H, t, J = 6.4 Hz) ESI-MS (+): 593.36 (M + H)

[0173] Example 2 Succinic acid {[(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1-yl)morpholin-2-yl]methyl}{2-octadecanoyloxy-1-[(octadecanoyloxymethyl)ethyl]} (hereinafter referred to as "G2-suc-morT-OFF").

[0174] Production of 4-((1,3-bis(stearoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (hereinafter referred to as "G2-suc"). To 1 g (1.60 mmol) of 2-hydroxypropane-1,3-diyl distearate, 8 mL of dichloromethane was added, 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 completion of the reaction, 1 M aqueous sodium dihydrogen phosphate solution was added to the reaction mixture, and the mixture was extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated to obtain G2-suc (1.40 g).

[0175] Production 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"). To 900 mg (1.24 mmol) of G2-suc and 277 mg (1.45 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 5.2 mL of dichloromethane was added, followed by 500 mg (1.03 mmol) of morT-OH and 132 mg (1.09 mmol) of 4-(N,N-dimethylamino)pyridine, and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, 0.1 M aqueous sodium dihydrogen phosphate solution was added to the reaction mixture, and the mixture was extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The obtained residue was purified by silica gel chromatography to obtain G2-suc-morT-ON (1.09 g, 89%).

[0176] 1 H-NMR (CDCl3): δ 8.04 (1H, s); 7.17 - 7.51 (15H, m); 6.98 (1H, s); 6.12 (1H, dd, J = 9.6 Hz, 2.4 Hz); 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 = 11.2 Hz); 3.10 (1H, d, J = 11.6 Hz); 2.60 (4H, s); 2.30 (4H, t, J = 7.6 Hz); 1.83 (3H, s); 1.38 - 1.44 (2H, m); 1.24 (60H, m); 0.87 (6H, t, J = 6.8 Hz)

[0177] Production of G2-suc-morT-OFF Dichloromethane (4.2 mL) was added to G2-suc-morT-ON, and the mixture was stirred at 0 °C. Then, 127 μL (0.62 mmol) of triisopropylsilane and 64 μL (0.82 mmol) of trifluoroacetic acid were added at 0 °C, and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, a saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with dichloromethane, dried over sodium sulfate, and the solvent was evaporated. The obtained residue was purified by silica gel chromatography to obtain G2-suc-morT-OFF (373 mg, 95%).

[0178] 1 H-NMR (CDCl3): δ 8.04 (1H, bs); 7.24 (1H, s); 5.70 (1H, d, J = 2 Hz); 5.21 - 5.26 (1H, m); 4.28 - 4.31 (2H, m); 4.13 - 4.17 (4H, m); 3.96 - 4.00 (1H, m); 3.11 (1H, dd, J = 12.4, 2 Hz); 2.94 (1H, dd, J = 12.8, 2.4 Hz); 2.57 - 2.65 (6H, m); 2.32 (4H, t, J = 7.6 Hz); 1.95 (3H, s); 1.25 (60H, m); 0.88 (6H, t, J = 7.6 Hz)

[0179] Example 3 [{(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl] succinic acid 1,3-bis(oleoyloxy)propan-2-yl (hereinafter referred to as "G3-suc-morT-OFF").

[0180] Production of [{(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl] succinic acid 1,3-bis(oleoyloxy)propan-2-yl (hereinafter referred to as "G3-suc-morT-ON"). Using 2-hydroxypropane-1,3-diyl dioleate as a raw material, 4-((1,3-bis(oleoyloxy)propan-2-yl)oxy)-4-oxobutanoic acid (hereinafter referred to as "G3-suc") was produced in the same manner as in Step 1 of Example 2. Subsequently, G3-suc-morT-ON was produced in the same manner as in Step 2 of Example 2.

[0181] 1 H-NMR (CDCl3): δ8.00 (1H, s); 7.17~7.51 (15H, m); 6.99 (1H, s); 6.09~6.12 (1H, m); 5.29~5.38 (4H, m); 5.20~5.25 (1H, m); 4.33~4.37 (1H, m); 4.26~4.30 (2H, m); 4.12~4.16 (2H, m); 4.00~4.09 (2H, m); 3.35 (1H, d, J = 11.6 Hz); 2.15 (1H, d, J = 11.6 Hz); 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 = 6.8 Hz)

[0182] Production of G3-suc-morT-OFF It was produced in the same manner as in Step 3 of Example 2.

[0183] 1 H-NMR (CDCl3): δ7.97 (1H, bs); 7.24 (1H, s); 5.69~5.72 (1H, m); 5.29~5.38 (4H, m); 5.21~5.25 (1H, m); 4.27~4.31 (2H, m); 4.13~4.17 (4H, m); 3.97~3.99 (1H, m); 3.11 (1H, d, J = 12 Hz); 2.94 (1H, d, J = 13.2 Hz); 2.57~2.67 (4H, m) 2.31 (4H, t, J = 7.6 Hz); 1.99~2.00 (11H, m); 1.26~1.29 (46H, m); 0.87 (6H, t, J = 6.8 Hz)

[0184] Example 4 4-Oxo-4-(4-stearoylpiperazin-1-yl)butanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl (hereinafter referred to as "G4-suc-morT-OFF"). Production of 4-oxo-4-(4-stearoylpiperazin-1-yl)butanoic acid (hereinafter referred to as "G4-suc") 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 were added to 26 mL of tetrahydrofuran. Then, 1.45 mL (10.7 mmol) of triethylamine and 1 g (5.37 mmol) of tert-butyl piperazine-1-carboxylate were added, and the mixture was stirred at room temperature for 16 hours. After completion of the reaction, a saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted with dichloromethane, dried over sodium sulfate, and concentrated under reduced pressure. The obtained residue was purified by silica gel chromatography to obtain tert-butyl 4-stearoylpiperazine-1-carboxylate (1.64 g; 67%). 18 mL of dichloromethane was added thereto, and the mixture was stirred at 0 °C. 2.77 mL (36.2 mmol) of trifluoroacetic acid was added at 0 °C, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, a saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted with dichloromethane, dried over sodium sulfate, and concentrated under reduced pressure to obtain 1-(piperazin-1-yl)octadecan-1-one (1.30 g). 18 mL of dichloromethane was added to 1.3 g (3.70 mmol) of the crude product, and then 0.41 g (4.10 mmol) of succinic anhydride and 0.77 mL (5.50 mmol) of triethylamine were added, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, acetone was added to the residue, and the mixture was slurried and washed at room temperature for 16 hours. The insoluble matter was collected by suction filtration, washed with acetone, and dried to obtain G4-suc (1.20 g).

[0185] Production of 4-oxo-4-(4-stearoylpiperazin-1-yl)butanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl (hereinafter referred to as "G4-suc-morT-ON") G4-suc (982 mg, 2.17 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (555 mg, 2.90 mmol) were added with tetrahydrofuran (10 mL), and the mixture was stirred at 70 °C. Subsequently, morT-OH (1 g, 2.07 mmol) and 4-(N,N-dimethylamino)pyridine (265 mg, 2.17 mmol) were added, and the mixture was stirred at 70 °C for 30 minutes. After completion of the reaction, the reaction mixture was allowed to cool to room temperature, 0.1 M aqueous sodium dihydrogen phosphate solution was added to the reaction solution, and the mixture was extracted with dichloromethane, dried over sodium sulfate, and the solvent was distilled off. The obtained residue was purified by silica gel chromatography to obtain G4-suc-morT-ON (1.68 g, 89%).

[0186] 1 1H-NMR (CDCl3): δ 8.00 (1H, s); 7.16 - 7.50 (15H, m); 6.97 (1H, s); 6.10 (1H, d, J = 8 Hz); 4.34 - 4.36 (1H, m); 4.04 (2H, d, J = 4.8 Hz); 3.57 - 3.64 (4H, m); 3.44 - 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 = 7.6 Hz); 1.82 (3H, s); 1.23 - 1.42 (32H, m); 0.86 (3H, t, J = 6.8)

[0187] Production of G4-suc-morT-OFF It was produced in the same manner as in Step 3 of Example 2.

[0188] 1 1H-NMR (CDCl3): δ 8.32 (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 = 7.2 Hz); 1.95 (3H, s); 1.25 -  1.31 (32H, m); 0.88 (3H, t, J = 7.6 Hz)

[0189] Example 5 4-(Octadecylcarbamoyl)benzoic acid [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl (hereinafter referred to as "G5-tpa-morT-OFF") Production of 4-(octadecylcarbamoyl)benzoic acid [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl]methyl (hereinafter referred to as "G5-tpa-morT-ON") It was produced in the same manner as in Step 2 of Example 2 using 4-(octadecylcarbamoyl)benzoic acid.

[0190] 1 H-NMR (CDCl3): δ 8.24 (1H, s); 7.97 (2H, d, J = 8 Hz); 7.77 (2H, d, J = 8 Hz); 7.17 - 7.46 (15H, m); 6.95 (1H, s); 6.12 - 6.16 (1H, m); 4.49 - 4.51 (1H, m); 4.25 - 4.33 (2H, m); 3.42 - 3.47 (2H, m); 3.35 - 3.38 (1H, m); 3.21 - 3.24 (1H, m); 1.79 (3H, s); 1.23 - 1.44 (34H, m); 0.86 (3H, t, J = 6.8 Hz)

[0191] Production of G5-tpa-morT-OFF It was produced in the same manner as in Step 3 of Example 2.

[0192] 1 H-NMR (CDCl3): δ 8.24 (1H, bs); 8.11 (2H, d, J = 8.4 Hz); 7.84 (2H, d, J = 8.4 Hz); 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 = 7.6 Hz)

[0193] Example 6 4-(4-(4-(Octadecylcarbamoyl)benzoyl)piperazin-1-yl)-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl (hereinafter referred to as "G6-suc-morT-OFF") Production of 4-(4-(4-(octadecylcarbamoyl)benzoyl)piperazin-1-yl)-4-oxobutanoic acid (hereinafter referred to as "G6-suc") Using 4-(octadecylcarbamoyl)benzoic acid instead of stearic acid, it was produced in the same manner as in Step 1 of Example 4.

[0194] Production of 4-[4-{4-(octadecylcarbamoyl)benzoyl}piperazin-1-yl]-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl (hereinafter referred to as "G6-suc-morT-ON") It was produced in the same manner as in Step 2 of Example 2.

[0195] 1 H-NMR (CDCl3): δ 8.08 (1H, bs); 7.81 (2H, d, J = 7.6 Hz); 7.16 - 7.50 (17H, m); 6.97 (1H, s); 6.08 - 6.10 (1H, m); 4.33 - 4.39 (1H, m); 4.02 - 4.04 (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 = 6.4 Hz)

[0196] Production of G6-suc-morT-OFF 4.6 mL of dichloromethane and 0.4 mL of 2,2,2-trifluoroethanol were added to 493 mg (0.47 mmol) of G6-suc-morT-ON493, and the mixture was stirred at 0 °C. Then, 145 μL (0.70 mmol) of triisopropylsilane and 53 μL (0.70 mmol) of trifluoroacetic acid were added at 0 °C, and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, a saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted with dichloromethane, dried over sodium sulfate, and the solvent was distilled off. The obtained residue was purified by silica gel chromatography to obtain G6-suc-morT-OFF (372 mg; 98%).

[0197] 1 H-NMR (CDCl3): δ 8.05 (1H, bs); 7.79 (2H, d, J = 7.6 Hz); 7.45 (2H, d, J = 7.6 Hz); 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 = 7.2 Hz)

[0198] Example 7 3,4,5-Tris(octadecyloxy)benzoic acid [(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl]methyl (hereinafter referred to as "G7-morT-OFF")

[0199] Production of 3,4,5-tris(octadecyloxy)benzoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-trityl-morpholin-2-yl}methyl (hereinafter referred to as "G7-morT-ON") It was produced in the same manner as in Step 2 of Example 2 using 3,4,5 - trioctadecyloxybenzoic acid.

[0200] 1 H-NMR (CDCl3): δ 7.90 (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 = 6.8 Hz)

[0201] Production of G7-morT-OFF It was produced in the same manner as in Step 3 of Example 2.

[0202] 1 H-NMR (CDCl3): δ 7.98 (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 = 7.2 Hz)

[0203] Example 8 Succinic acid {(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) (hereinafter referred to as "G8-suc-morT-OFF")

[0204] Step 1 Production of 2-hydroxyethyl 3,4,5-tri(octadecyloxy)benzoate 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. Then, 120 mg (1.90 mmol) of ethylene glycol was added, and the mixture was stirred at room temperature for 3 hours. After completion of the reaction, a 1 M aqueous solution of sodium dihydrogen phosphate was added to the reaction solution, and the mixture was extracted with dichloromethane, dried over sodium sulfate, and the solvent was distilled off. The obtained residue was purified by silica gel chromatography to obtain 2-hydroxyethyl 3,4,5-trioctadecyloxybenzoate (882 mg; 56%)

[0205] 1 1H-NMR (CDCl3): δ 7.26 (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 = 7.2 Hz)

[0206] Step 2 Production of succinic acid {(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) (hereinafter referred to as "G8-suc-morT-ON") In the same manner as in Step 1 of Example 2, 4-oxo-4-(2-[{3,4,5-tris(octadecyloxy)benzoyl}oxy]ethoxy)butanoic acid (hereinafter referred to as "G8-suc") was obtained, and then G8-suc-morT-ON was obtained in the same manner as in Step 2 of Example 2.

[0207] 1 1H-NMR (CDCl3): δ 7.87 (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 = 7.2 Hz)

[0208] Step 3 Production of G8-suc-morT-OFF G8-suc-morT-OFF was obtained in the same manner as in Step 3 of Example 2.

[0209] 1 H-NMR (CDCl3): δ 7.96 (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 = 6.8 Hz)

[0210] Example 9 4-(Dioctadecylamino)-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl (hereinafter referred to as "G9-suc-morT-OFF") Step 1 Production of 4-(dioctadecylamino)-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl (hereinafter referred to as "G9-suc-morT-ON") Using N-octadecyl octadecane-1-amine as a raw material, 4-(dioctadecylamino)-4-oxobutanoic acid (hereinafter referred to as "G9-suc") was produced in the same manner as in Step 1 of Example 2. Subsequently, G9-suc-morT-ON was produced in the same manner as in Step 2 of Example 2.

[0211] 1 H-NMR (CDCl3): δ 7.88 (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 = 6.8 Hz)

[0212] Step 2 Production of G9-suc-morT-OFF It was produced in the same manner as in Step 3 of Example 2.

[0213] 1 H-NMR (CDCl3): δ 8.14 (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 = 7.2 Hz)

[0214] Example 10 4-[{1-(octadecylamino)-1-oxo-3-phenylpropan-2-yl}amino]-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl}methyl (hereinafter referred to as "G10-suc-morT-OFF") Production of 4-[(1-octadecylamino-1-oxo-3-phenylpropan-2-yl)amino]-4-oxobutanoic acid {(2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl}methyl (hereinafter referred to as "G10-suc-morT-ON") in Step 1 To 500 mg (1.88 mmol) of 2-tert-butoxycarbonylamino-3-phenyl-propanoic acid was added 9.4 mL of tetrahydrofuran, followed by addition of 652 μL (3.77 mmol) of N-ethyl-N-isopropyl-propan-2-amine, 46 mg (0.38 mmol) of 4-(N,N-dimethylamino)pyridine, 505 mg (2.64 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, and octadecane-1-amine, and the mixture was stirred at room temperature for 5 hours. After completion of the reaction, 1 M aqueous sodium dihydrogen phosphate solution was added to the reaction solution, and the mixture was extracted with dichloromethane, dried over sodium sulfate, and the solvent was distilled off. The obtained residue was purified by silica gel chromatography to obtain tert-butoxycarbonylamino-N-octadecyl-3-phenyl-propaneamide (779 mg, 80%)

[0215] 1 H-NMR (CDCl3): δ 7.18~7.28 (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 = 6.8 Hz)

[0216] To 779 mg (1.51 mmol) of tert-butoxycarbonylamino-N-octadecyl-3-phenyl-propaneamide was added 15 mL of dichloromethane, followed by addition of 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-propaneamide (620 mg). The same reaction as in Step 1 of Example 2 was carried out using the crude product as it was to produce G-10-suc. Then, the same reaction as in Step 2 of Example 2 was carried out to produce G-10-suc-morT-ON.

[0217] 11H-NMR (CDCl3): δ 8.08 (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 = 5.6 Hz); 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 = 7.6 Hz)

[0218] Production of G10-suc-morT-OFF in Step 2 The target product was obtained in the same manner as in Step 3 of Example 2.

[0219] 1 1H-NMR (CDCl3): δ 8.33 (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 = 7.6 Hz)

[0220] The chemical structural formulas of the compounds described in Examples 1 to 10 above are shown in Table 5 below.

[0221] [Table 5]

[0222] Production of Example 11 ((2S,6R)-6-(4-Benzamido-2-oxopyrimidin-1(2H)-yl)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)morpholin-2-yl)methyl 4-(octadecylamino)-4-oxobutanoate (hereinafter referred to as G1-suc-PMO[C-T]-ON(3))

Chemical formula

[0223] Example 12 Preparation of ((2S,6R)-4-((Dimethylamino)(((2S,6R)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methoxy)phosphoryl)-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-PMO[T-T-T]-ON(5)) [Chemical formula] 2 mL of solution a shown below, delivered at a flow rate of 0.5 mL / min, and 2 mL of solution b shown below, delivered at a flow rate of 0.5 mL / min, were mixed in a static mixer and then reacted at room temperature for 0.4 minutes in a 0.4 mL tube reactor. Thereafter, 2 mL of solution c shown below, delivered at a flow rate of 0.5 mL / min, was mixed and reacted at room temperature for 3.3 minutes in a 5 mL tube reactor. Of the resulting solution, 100 μL was mixed with 200 μL of a 0.02 M iodine solution (THF / water = 99.8 / 0.2, v / v) and stirred at room temperature for 15 minutes. After the reaction was completed, a 10% aqueous sodium thiosulfate solution was added to the reaction solution. After separating the organic layer and the aqueous layer, the organic layer was diluted 10-fold with acetonitrile and analyzed by HPLC (starting material Rt: 12.69 min, target product Rt: 15.35 min, conversion yield 90.4%). Solution a: 40 μL (0.35 mmol) of dichloro(dimethylamino)phosphine was dissolved in 2.5 mL of dichloromethane. Solution b: 169 mg (0.35 mmol) of 1 - ((2R,6S)-6-(hydroxymethyl)-4-tritylmorpholin-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (1) and 137 μL (0.788 mmol) of N,N-diisopropylethylamine were dissolved in 2.5 mL of dichloromethane. Solution c: 162 mg (0.175 mmol) of G1-suc-PMO[T―T]-OFF(4) and 76 μL (0.438 mmol) of N,N-diisopropylethylamine were dissolved in 2.5 mL of dichloromethane.

[0224] In Examples 13 to 15, the following analysis conditions were used. <ODS conditions> Column: Waters XBridge C8 (5 μm, 4.6×75 mm), 60 °C Detection wavelength: 264 nm Mobile phase A: 50 mM TEAAaq. Mobile phase B: MeOH Flow rate: 0.75 mL / min Gradient: 70 - 95% B (0 - 20 min) 95% B (20 - 26 min) 75% B (26 - 35 min)

[0225] Example 13 Preparation of ((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-(((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)morpholin-2-yl)methyl 4-(octadecylamino)-4-oxobutanoate (hereinafter referred to as G1-suc-PMO[A Bz -A Bz -ON(8))

Chemical formula

[0226] Example 14 Preparation of ((2S,6R)-6-(6-benzamido-9H-purin-9-yl)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)morpholin-2-yl)methyl 4-(octadecylamino)-4-oxobutanoate (hereinafter referred to as G1-suc-PMO[A Bz ―T]-ON(9))

Chemical formula

[0227] Example 15 Preparation of ((2S,6R)-4-((dimethylamino)(((2S,6R)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-4-tritylmorpholin-2-yl)methoxy)phosphoryl)-6-(5-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)morpholin-2-yl)methyl 4-(octadecylamino)-4-oxobutanoate (hereinafter, G1-suc-PMO[T―T]-ON(11)) [Chemical formula] 2 mL of solution a shown below delivered at a flow rate of 0.5 mL / min and 2 mL of solution b shown below delivered at a flow rate of 0.5 mL / min were mixed in a static mixer and then reacted at room temperature for 2 minutes in a 2 mL tube reactor. Thereafter, 2 mL of solution c shown below delivered at a flow rate of 1 mL / min was mixed and reacted at room temperature for 6.7 minutes in a 10 mL tube reactor. 100 μL of the resulting solution was mixed with 200 μL of a 0.02 M iodine solution (THF / water = 99.8 / 0.2, v / v) and stirred at room temperature for 15 minutes. After completion of the reaction, a 10% aqueous sodium thiosulfate solution was added to the reaction solution. After separating the organic layer and the aqueous layer, the organic layer was diluted 10-fold with acetonitrile and analyzed by HPLC (starting material Rt: 12.21 min, target product Rt: 18.10 min, conversion yield 96.5%). Solution a: 38 μL (0.333 mmol) of dichloro(dimethylamino)phosphine was dissolved in 2.4 mL of dichloromethane. Solution b: 105 mg (0.216 mmol) of 1-((2R,6S)-6-(hydroxymethyl)-4-tritylmorpholin-2-yl)-5-methylpyrimidine-2,4(1H,3H)-dione (1) and 95 μL (0.549 mmol) of N,N-diisopropylethylamine were dissolved in 2.4 mL of dichloromethane. Solution c: 100 mg (0.166 mmol) of G1-suc-morT-OFF(10) and 72 μL (0.416 mmol) of N,N-diisopropylethylamine were dissolved in 2.4 mL of dichloromethane.

Claims

1. General formula [B-1-1]: 【Chemical Formula 1】 [In the formula, B Ｐ each independently represents a nucleic acid base which may be protected; p represents an integer from 1 to 10; Q １ represents a group removable under acidic conditions; W each represents the same or different and represents a lone pair of electrons, an oxygen atom, or a sulfur atom; and X is, each independently the same or different, a hydroxyl group substituted with a group capable of leaving 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 １－６ alkyl)amino substituted with a group capable of leaving under basic conditions, di(amino-C １－６ alkyl)amino substituted with a group capable of leaving under basic conditions or the following general formula [2]: [Chemical Formula 2] (In the formula, * represents the bonding position with P; a represents an integer from 0 to 2; E is CH ２ , CH-A １ or N-A 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 removable under basic conditions, mono(C １－６ alkyl)amino substituted with a group removable under basic conditions, 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]: [Chemical Formula 3] In the formula, * represents the bonding position with E; b represents an integer from 0 to 2; c represents 0 or 1; R １１ represents C １－６ alkyl; and M represents CH ２ , an oxygen atom, a sulfur atom or N-(a group removable under basic conditions).) represents a substituent represented by; and A ２ is C １－６ alkyl, mono(C １－６ alkyl)amino-C １－６ alkyl, di(C １－６ alkyl)amino-C １－６ alkyl, tri(C １－６ alkyl)ammonio-C １－６ alkyl, a group removable under basic conditions, aryl or heteroaryl. ) represents a substituent represented by.] The compound represented by is made into a compound of general formula [P]: 【Chemical Formula 4】 [wherein, X is as defined above, LG １ and LG ２ are the same or different and are a leaving group, for example, a halogen (chloro, bromo, iodo, particularly chloro).] is reacted with a compound of general formula [B-1-2]: 【Chemical Formula 5】 [wherein, p, B Ｐ , Q １ , W, X, and LG １ are as defined above.]; the step of forming a compound of Step 2) The compound of the general formula [B-1-2] is General formula [A-1]: [Chemical Formula 6] [In the formula, B Ｐ , W and X have the same meanings as described above, G is (1) a silicon substituent, (2) a long-chain alkyl-carbonyl, (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy or (4) the following general formula [7]: 【Chemical Formula 7】 (In the formula, * represents the bonding position with T; Z is (1) (a soluble polymer dissolved in an organic solvent)-oxy, (2) (a soluble polymer dissolved in an organic solvent)-amino, (3) long-chain alkyloxy, (4) a solid support or (5) the following general formulas [8A] to [8N]: 【Chemical 8】 (In the formula, * represents the bonding position with L; j represents an integer from 0 to 4; k represents an integer from 0 to 5; R ８ａ represents a hydrogen atom or C １－６ alkyl; R ８ｂ each independently represents a long-chain alkyl group; R ８ｃ are each the same or different and are represented by the following general formulas [9A] to [9E]: 【Chemical Formula 9】 (In the formula, * represents the bonding position; and R ９ represents a long-chain alkyl and / or long-chain alkenyl.) represents a substituent represented by; R ８ｄ each independently represents a hydrogen atom, a halogen, a long-chain alkyl which may be substituted with 1 to 13 halogens, or a long-chain alkyloxy which may be substituted with 1 to 13 halogens; R ８ｅ is (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) benzoyl substituted with 1 to 5 long-chain alkyloxy and / or long-chain alkenyloxy represents; and R ８ｆ is (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) long-chain alkenyl-carbonyl represents.) represents a substituent represented by; and L is general formula [10]: 【Chemical 10】 (In the formula, * represents the bonding position with Z; ** represents the bonding position with T; and L １ is an optionally substituted C ２－１０ alkylene or an optionally substituted C ６－１０ arylene. ) represents a substituent represented by.) represents a substituent represented by; n represents an integer from 1 to 50; and T is a single bond or the following general formula [11]: 【Chemical Formula 11】 (In the formula, X and W have the same meanings as described above; * represents the bonding position with **O, *O or *N in the above general formulas [4a] to [4d]; ** represents the bonding position with G; and q represents an integer from 0 to 10.) represents a substituent represented by, provided that when G is a silicon substituent, T is a single bond.] react with the compound represented by to General formula [C-1-1]: 【Chemical 12】 [wherein, n, p, B Ｐ , Q １ , G, T, W, and X are as defined above.] the step of forming a compound represented by A method for producing a compound represented by general formula [C-1-1], comprising

2. General formula [A-1]: 【Chemical 13】 [wherein, n, B Ｐ , G, T, W, and X are as defined in claim 1.] The compound represented by is of general formula [P]: 【Chemical Formula 14】 [wherein, X, LG １ , and LG ２ are as defined in claim 1.] React with a compound of the general formula [A-1-3]: 【Chemical Formula 15】 [wherein, n, B Ｐ , W, X, G, T, and LG １ are as defined in claim 1.] The step of forming the compound, and Step 2') Reacting the compound of the general formula [A-1-3] with a compound represented by the general formula [B-1-1]: 【Chemical Formula 16】 [wherein, p, B Ｐ , W, X, Q １ are as defined in claim 1.] to form a compound represented by the general formula [C-1-1]: 【Chemical 17】 [wherein, n, p, B Ｐ , Q １ , G, T, W, and X are as defined above.] The step of forming the compound A method for producing a compound represented by the general formula [C-1-1], comprising:

3. Furthermore, treating the compound of the general formula [C-1-1] formed by the method according to claim 1 or claim 2 with an oxidizing agent, to obtain a compound represented by the general formula [C-1]: 【Chemical 18】 [wherein n, p, B Ｐ , Q １ , G, T, W, and X are as defined in claims 1 and 2.], the production method according to claim 1 or claim 2, which comprises a step of forming a compound represented by

4. Furthermore, a solution containing an acid is added to a reaction mixture containing the compound [C-1] formed by the method of claim 3, and the removal reaction of Q １ is continuously carried out in the same system to obtain a compound of the general formula [E-1]: 【Chemical Formula 19】 [wherein, n, p, B Ｐ , Q １ , G, T, W, and X are as defined in claims 1 and 2.], the production method according to claim 1 or claim 2, which comprises forming a compound represented by the formula.

5. Furthermore, a method according to claim 1 or 2, comprising the step of forming a compound of the general formula [A-1-1]: 【Chemical 20】 [wherein, n, B Ｐ , Q １ , G, T, W, and X are as defined in claims 1 and 2.], from the compound of], Q １ is removed to obtain A compound of the general formula [A-1]: 【Chemical 21】 [wherein, B Ｐ , W, X, G, T, and n are as defined above.]

6. A compound of the general formula [A-1-1]: 【Chemical 22】 [wherein, n, B Ｐ , G, T, W, and X are as defined in claims 1 and 2.], from the compound of Q １ is removed to obtain A compound of the general formula [A-1]: 【Chemical 23】 [wherein, B Ｐ , W, X, G, T, and n are as defined above.], to form a compound of the formula [A-1], and the compound of the general formula [A-1], A compound of the general formula [B-1]: 【Chemical formula 24】 [wherein, B Ｐ , Q １ , W, X, G, and T are as defined above; D is a halogen, p is an integer from 1 to 10] and reacting with a compound of to obtain a compound represented by the general formula [C-1]: 【Chemical Formula 25】 [wherein, n, p, B Ｐ , Q １ , W, X, G, and T are as defined above.], a method for producing a compound represented by the general formula [C-1], which comprises forming a compound represented by the formula.

7. Supplying a solution containing the compound of the general formula [A-1] and a solution containing the compound of the general formula [B-1] to a flow reactor to form the compound of the general formula [C-1], and optionally Supplying a solution containing the compound of the general formula [C-1] and a solution containing an acid to a flow reactor to remove Q １ and forming a compound of the formula [E-1], which comprises The method according to claim 6.

8. A solution containing the compound of the general formula [A-1-1] and a solution containing an acid are fed into a flow reactor to remove Q １ to form the compound of the formula [A-1], and then Supplying a solution containing the compound of the general formula [A-1] and a solution containing the compound of the general formula [B-1] to the following flow reactor to form the compound of the general formula [C-1], comprising The method according to claim 6.

9. A method for producing a compound represented by the general formula [B-0-1]: 【Chemical 26】 [wherein, B Ｐ and Q １ are as defined in claim 1.] Reacting a compound of the general formula [B-0] with a compound of the general formula [P]: 【Chemical 27】 [wherein, X, LG １ , and LG ２ are as defined in claim 1.] 【Chemical formula 28】 [wherein, B Ｐ , Q １ , X, and LG １ are as defined above.]

10. A compound of the general formula [B-0-1] 【Chemical 29】 [wherein, B Ｐ , Q １ , X, and LG １ are as defined in claim 1.] Reacting with A compound of the general formula [A-0]: 【Chemical 30】 [wherein, B Ｐ , G, and T have the same meanings as in claim 1.] to obtain a compound represented by the general formula [C-0-1]: 【Chemical 31】 [In the formula, B Ｐ , Q １ , G, T, and X have the same meanings as described above.] A method for producing a compound.

11. A compound of the general formula [C-0-1] 【Chemical 32】 [wherein, B Ｐ , Q １ , G, T, and X are as defined in claims 1 and 2.] A method for preparing a compound represented by the general formula [C-0], comprising treating with an oxidizing agent. 【Chemical 33】 [wherein, B Ｐ , Q １ , G, T, W, and X are as defined above]

12. B Ｐ is an optionally protected nucleobase; Q １ is a group that can be eliminated under acidic conditions; W is a lone pair, an oxygen atom or a sulfur atom; X is di(C １－６ alkyl)amino or has the general formulas [2-1] to [2-8]: 【Chemical Formula 34】 [wherein, * represents the bonding position with P] Selected from substituents represented by G is a general formula [7]: 【Chemical 35】 (wherein, * represents the bonding position with T; Z is a general formula [8A] to [8D], [8E], [8G], [8H], [8J], [8K], [8N]: 【Chemical 36】 (wherein, * represents the bonding position with L; k represents an integer from 0 to 5; R ８ａ represents a hydrogen atom or C １－６ alkyl; R ８ｂ each independently represents a long-chain alkyl group; R ８ｃ are each the same or different and have the following general formula [9A]: 【Chemical 37】 (wherein, * represents the bonding position; and R ９ represents a long-chain alkyl and / or a long-chain alkenyl.) represents a substituent represented by; R ８ｄ each independently represents a hydrogen atom, a halogen, a long-chain alkyl which may be substituted with 1 to 13 halogens, or a long-chain alkyloxy which may be substituted with 1 to 13 halogens; R ８ｅ is (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) represents one to five benzoyls substituted with long-chain alkyloxy and / or long-chain alkenyloxy; and R ８ｆ is (1) long-chain alkyl, (2) long-chain alkyl-carbonyl or (3) long-chain alkenyl-carbonyl. ) is a substituent represented by; L is of the general formula [10]: 【Chemical 38】 (wherein, * represents the bonding position with Z; ** represents the bonding position with the oxygen atom; and L １ represents an optionally substituted C ２－１０ alkylene or an optionally substituted C ６－１０ arylene. ) is a substituent represented by. ) is selected from the substituents represented by; T is a single bond or the following general formula [11]: 【Chemical 39】 (wherein, X and W have the same meanings as described above; * represents the bonding position with O; ** represents the bonding position with G; and q represents an integer from 0 to 10. ) is a substituent represented by, n is from 1 to 25. ] The production method according to any one of claims 1 to 11.

13. B Ｐ is an optionally protected nucleobase; Q １ is H or a group that is removable under acidic conditions; W is an oxygen atom; X is di(C １－６ alkyl)amino; G is of the following formula: 【Chemical Formula 40】 [wherein, * represents the bonding position with T. ] is selected from the group consisting of; T is a single bond; n is from 1 to 25. ] The production method according to any one of claims 1 to 12.

14. Q １ is trityl, monomethoxytrityl, or dimethoxytrityl, and n, B Ｐ , W, X, G, and T are as defined above. from the compound of, in the presence of trifluoroacetic acid and 2,2,2-trifluoroethanol, and optionally triisopropylsilane or ethanol, Q １ including removing The production method according to any one of claims 4 to 13.

15. The method according to any one of claims 1 to 14, wherein the optionally protected nucleobases are each independently adenine, guanine, hypoxanthine, cytosine, thymine, uracil or modified bases thereof.

16. The protected nucleobase is a nucleobase in which the amino group or hydroxyl group of the nucleobase is protected by a protecting group, and the protecting group for the amino group is independently selected from the group consisting of benzoyl, 4-methoxybenzoyl, acetyl, propionyl, butyryl, isobutyryl, phenylacetyl, phenoxyacetyl, 4-tert-butylphenoxyacetyl, 4-isopropylphenoxyacetyl, (dimethylamino)methylene, and the protecting group for the hydroxyl group is independently selected from the group consisting of 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, 4-(phenylcarboxy)benzyl, the method according to any one of claims 1 to 14.

17. G is of the following formula: 【Chemical Formula 41】 [wherein, * represents the bonding position with T. ] selected from the group consisting of; The method according to any one of claims 1 to 16, wherein T is a single bond.

18. wherein X is di(C １－６ alkyl)amino, the method according to any one of claims 1 to 17.

19. LG １ and LG ２ The method according to any one of claims 1 to 18, wherein X is the same as or different from, and is a halogen.

20. The method according to any one of claims 1 to 19, wherein W is an oxygen atom.

21. The method according to any one of claims 3 to 19, wherein the oxidizing agent is iodine or magnesium monoperoxyphthalate hexahydrate.

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