Method for producing alkyl groups

Abstract

To provide a method for producing oligonucleotides that can suppress the formation of by-products. [Solution] A method for producing an oligonucleotide having a phosphate ester bond by oxidizing an oligonucleotide precursor having a phosphite ester bond or a phosphonic acid ester bond with an oxidizing agent, wherein the oxidizing agent is a compound represented by formula (I) (excluding 2,2-dipyridyl disulfide) (the definition of the symbols in the formula is as described in the specification). JPEG2026110729000086.jpg844

Description

[Technical Field]

[0001] This invention relates to a method for producing oligonucleotides. [Background technology]

[0002] The phosphoramidite method and the H-phosphonate method are widely used as methods for producing oligonucleotides. In the phosphoramidite method, which involves oxidation, for example, as shown in the following formula, a phosphoramidite (compound (1)) and a nucleoside (compound (2)) are condensed to synthesize an oligonucleotide precursor (compound (3)) having a phosphite ester bond, and then the oligonucleotide precursor is oxidized to obtain an oligonucleotide (compound (4)). In this specification, "compound represented by formula (1)" may be abbreviated as "compound (1)". Compounds represented by other formulas, groups represented by other formulas, etc. may also be abbreviated in the same way.

[0003] [ka]

[0004] [In the formula, DMTr represents a 4,4'-dimethoxytrityl group, i-Pr represents an isopropyl group, CE represents a 2-cyanoethyl group, Base and Base' represent optionally protected nucleic acid bases, and R represents a protecting group or solid support.]

[0005] In the phosphoramidite method, nucleotides or oligonucleotides (for example, oligonucleotides having a thiophosphate ester bond) may be condensed instead of nucleosides. Furthermore, in the phosphoramidite method, sulfurization of the oligonucleotide precursor may be performed instead of oxidation of the oligonucleotide precursor.

[0006] In the H-phosphonate method, which involves oxidation, an activated H-phosphonate (compound (5)) is synthesized by reacting it with pivaloyl chloride (PivCl), as shown in the following formula. This activated H-phosphonate is then condensed with a nucleoside (compound (7)) to synthesize an oligonucleotide precursor (compound (8)) having a phosphonic acid ester bond. This oligonucleotide precursor is then oxidized to obtain an oligonucleotide (compound (9)).

[0007] [ka]

[0008] [In the formula, DMTr represents 4,4'-dimethoxytrityl, N(Et)3 represents triethylamine, Base and Base' represent optionally protected nucleic acid bases, Piv represents a pivaloyl group, and R represents a protecting group or solid support.]

[0009] In the H-phosphonate method, nucleotides or oligonucleotides (for example, oligonucleotides having a thiophosphate ester bond) may be condensed instead of nucleosides. Furthermore, in the H-phosphonate method, sulfurization of the oligonucleotide precursor may be performed instead of oxidation of the oligonucleotide precursor.

[0010] In addition to the representative phosphoramidite and H-phosphonate methods mentioned above, various other methods for producing oligonucleotides have been proposed. For example, Non-Patent Literature 1 discloses the production of a dinucleotide (compound 4) by condensing and oxidizing an H-phosphonate (compound 1) and a nucleoside (compound 2) in the presence of a large amount of triphenylphosphine and a large amount of 2,2'-dipyridyl disulfide, as shown in the formula below.

[0011] [ka]

[0012] [In the formula, DMTr represents 4,4'-dimethoxytrityl, Th represents a thymine base, PPh3 represents triphenylphosphine, and Ac represents an acetyl group.]

[0013] Furthermore, although Non-Patent Document 2 and Patent Document 1 do not disclose a method for producing oligonucleotides, they disclose the reduction of a benzoisothiazolone derivative (compound 1) with triethyl phosphite (P(OEt)3) as shown in the formula below (in other words, the oxidation of triethyl phosphite (P(OEt)3) with a benzoisothiazolone derivative (compound 1) to form triethyl phosphate (O=P(OEt)3)).

[0014] [ka] [Prior art documents] [Patent Documents]

[0015] [Patent Document 1] International Publication No. 2017 / 070157 [Non-patent literature]

[0016] [Non-Patent Document 1] ARKIVOC 2009 (iii) 264-273 [Non-Patent Document 2] J. Am. Chem. Soc. 2016, 138, 6715-6718 [Overview of the Initiative] [Problems that the invention aims to solve]

[0017] In the phosphoramidite method, iodine is widely used as an oxidizing agent for oligonucleotide precursors having phosphite ester bonds (for example, compounds represented by formula (3) above). Similarly, in the H-phosphonate method, iodine is widely used as an oxidizing agent for oligonucleotide precursors having phosphonic acid ester bonds (for example, compounds represented by formula (8) above).

[0018] In this regard, the present inventors have found that when iodine is used as an oxidizing agent for oligonucleotide precursors having a phosphite ester bond, a by-product (hereinafter sometimes referred to as "deficient product") is formed by the decomposition of the phosphite ester bond site of the precursor in reaction with the oxidizing agent. Furthermore, they have found that when oligonucleotide precursors having a thiophosphate ester bond in addition to a phosphite ester bond or phosphonic acid ester bond are oxidized with iodine, a by-product (hereinafter sometimes referred to as "desulfurized product") is formed by the conversion of the thiophosphate ester bond to a phosphate ester bond.

[0019] The present invention has been made in view of the circumstances described above, and its purpose is to provide a method for producing oligonucleotides that can suppress the formation of the aforementioned defective or desulfurized products compared to the use of iodine. [Means for solving the problem]

[0020] As a result of diligent research by the inventors, formula (I):

[0021] [ka]

[0022] [In the formula, X 1 This is a single bond, sulfur atom, oxygen atom, -S(=O)2- or -N(-R 3 )- indicates, X 1 If it is a single bond, R 1represents a halogen atom, and R 2 represents an optionally substituted aryl group or an optionally substituted heteroaryl group, X 1 when is a sulfur atom or -S(=O)2-, R 1 represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 represents an optionally substituted aryl group or an optionally substituted heteroaryl group, X 1 when is an oxygen atom, R 1 and R 2 together with the sulfur atom and oxygen atom to which they are attached, form an optionally substituted heterocyclic ring, and X 1 when is -N(-R 3 )-, R 1 represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 and R 3 together with the sulfur atom and nitrogen atom to which they are attached, form an optionally substituted heterocyclic ring, or R 1 and R 3 together with the nitrogen atom to which they are attached, form an optionally substituted heterocyclic ring, and R 2 represents an optionally substituted aryl group or an optionally substituted heteroaryl group.] It has been found that by using the compound represented by (excluding 2,2-dipyridyl disulfide) as an oxidizing agent to oxidize an oligonucleotide precursor having a phosphite ester bond or a phosphonate ester bond, the formation of the aforementioned by-products can be suppressed. The present invention based on this finding is as follows.

[0023] [1] A method for producing an oligonucleotide having a phosphate ester bond by oxidizing an oligonucleotide precursor having a phosphite ester bond or a phosphonate ester bond with an oxidizing agent, The oxidizing agent is defined by formula (I):

[0024] [ka]

[0025] [In the formula, X 1 This is a single bond, sulfur atom, oxygen atom, -S(=O)2- or -N(-R 3 )- indicates, X 1 If it is a single bond, R 1 R indicates a halogen atom, and 2 This represents an optionally substituted aryl group or an optionally substituted heteroaryl group. X 1 If R is a sulfur atom or -S(=O)2-, 1 R represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 This represents an optionally substituted aryl group or an optionally substituted heteroaryl group. X 1 If R is an oxygen atom, 1 and R 2 Together with the sulfur and oxygen atoms to which they are bonded, they form a heterocycle which may be substituted, and X 1 -N(-R 3 )- If R 1 R represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 and R 3 They, together with the sulfur and nitrogen atoms to which they bond, form a heterocycle which may be substituted, or R 1 and R 3 Together with the nitrogen atoms to which they are bonded, they form a heterocycle which may be substituted, and R 2 This indicates an optionally substituted aryl group or an optionally substituted heteroaryl group. A method using a compound represented by (excluding 2,2-dipyridyl disulfide).

[0026] [2] The compound represented by formula (I) is formula (Ia):

[0027] [ka]

[0028] [In the formula, R 1a This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Ring A exhibits a 5- or 6-membered unsaturated heterocycle. Y 1a This represents -S(=O)-, -S(=O)2-, -C(=O)-, or -C(=S)-. m represents an integer between 0 and 3. m R 2a Each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and If m is an integer greater than or equal to 2, then two adjacent R 2a It may form a fused ring, which may be substituted, together with ring A. Compounds represented by formula (Ib):

[0029] [ka]

[0030] [In the formula, R 1b This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Ring B represents a 6-membered aromatic hydrocarbon ring, or a 5- or 6-membered aromatic heterocycle. n represents an integer from 0 to 5. n R 2bEach independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and If n is an integer greater than or equal to 2, then two adjacent R 2b It may form a substituted or bicyclic fused aromatic heterocycle together with ring B. Compounds represented by (excluding 2,2-dipyridyl disulfide) The method described in [1] above.

[0031] [3] The compound represented by formula (Ia) is, formula (Ic):

[0032] [ka]

[0033] [In the formula, R 1c This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Y 1c This represents -S(=O)-, -S(=O)2-, -C(=O)-, or -C(=S)-. p represents 0 or 1. R 2c This represents a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. Ring C represents a 6-membered aromatic hydrocarbon ring, a 6-membered nitrogen-containing aromatic heterocycle, or a 10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom. q represents an integer from 0 to 4, and q R 3c Each of these independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. The method according to [2] above, wherein the compound is represented by .

[0034] [4] The compound represented by formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazole Lu-3(2H)-one, 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one, 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3 (2H)-one, 2-phenylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one, 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-phenyl-2H-1,2-benzothiadin-3(4H)-one, 2-(2,6-dimethylphenyl)isothiazolo[5,4- The method according to [3], wherein at least one is selected from the group consisting of [b]pyridine-3(2H)-one, 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one, 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one, and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one.

[0035] [5] The method according to [3], wherein the ring C is a 6-membered aromatic hydrocarbon ring or a 6-membered nitrogen-containing aromatic heterocycle. [6] The compound represented by formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazole-3(2H)-one, 2-(2,6-dimethyl The method according to [5], wherein at least one selected from the group consisting of phenyl)-5-nitro-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one, and 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3(2H)-one.

[0036] [7] Formula (r1) in the compound represented by formula (Ib):

[0037] [ka]

[0038] The base represented by, (1) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (2) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (6) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (7) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (8) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (9) A phenyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (10) A 2-imidazolyl group which may be substituted with 1 to 3 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. The method described in any one of the above [2] to [6].

[0039] [8] The method according to [7], wherein the compound represented by formula (Ib) is at least one selected from the group consisting of 2,2'-dibenzothiazolyl disulfide, 2-(2-benzothiazolyldithio)propanoic acid, 3-(2-benzothiazolyldithio)propanoic acid, 2,2'-dibenzimidazolyl disulfide, 2,2'-dibenzoxazolyl disulfide, 5,5'-di(1,2,3-triazolyl) disulfide, 5,5'-dithiobis(1-phenyl-1H-tetrazole), 2,2'-dithiobis(pyridine-N-oxide), 4,4'-dipyridyl disulfide, 3-(2-pyridyldithio)propanoic acid, 2,2'-dithiobis(1H-imidazole), and 2,2'-dithiobis(1-methyl-1H-imidazole).

[0040] [9] The method according to any one of the above [1] to [8], wherein the oxidation is carried out in the presence of water.

[0041]

[10] Oligonucleotide precursors having a phosphite ester bond or a phosphonic acid ester bond, (1) A phospholite ester bond-containing oligonucleotide precursor obtained by condensation of a nucleoside, nucleotide, or oligonucleotide with a phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide, or (2) Oligonucleotide precursors having phosphite bonds obtained by condensation of oligonucleotides with phosphoramidites The method described in any one of the above [1] to [9].

[0042]

[11] The method according to any one of [1] to

[10] , wherein the oligonucleotide precursor having a phosphite ester bond or a phosphonate ester bond is an oligonucleotide precursor having a thiophosphate ester bond in addition to the phosphite ester bond or phosphonate ester bond.

[0043]

[12] A method for producing oligonucleotides by one-pot synthesis, The aforementioned one-pot synthesis, Step (1) involves condensing a nucleoside, nucleotide, or oligonucleotide (a) having a hydrophobic protecting group with a nucleoside, nucleotide, or oligonucleotide (b) that is protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions and is phosphoramidized, in a solution containing a nonpolar solvent, to form an oligonucleotide precursor (c) having a phosphite ester bond and a hydrophobic protecting group, and protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions. Step (2) involves adding a quenching agent (i) for the phosphoramidized nucleoside, nucleotide, or oligonucleotide (b) to the solution after step (1) to quench the phosphoramidized nucleoside, nucleotide, or oligonucleotide (b). Step (3) involves adding an oxidizing agent to the solution after step (2) to oxidize the oligonucleotide precursor (c) to form an oligonucleotide (d) having a phosphate ester bond and a hydrophobic protecting group, and being protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions. Step (4) involves adding an oxidizing agent quenching agent (ii) to the solution after step (3) to quench the oxidizing agent. Step (5) involves adding an acid to the solution after step (4) to remove the temporary protecting group of the oligonucleotide (d) that can be removed under acidic conditions, thereby forming an oligonucleotide (e) having an unprotected hydroxyl group and a hydrophobic protecting group, and (7) A step in which a polar solvent is added to the solution containing the oligonucleotide (e) to precipitate the oligonucleotide (e). Including, and The oxidizing agent is defined by formula (I):

[0044] [ka]

[0045] [In the formula, X 1 This is a single bond, sulfur atom, oxygen atom, -S(=O)2- or -N(-R 3)- indicates, X 1 If it is a single bond, R 1 R indicates a halogen atom, and 2 This represents an optionally substituted aryl group or an optionally substituted heteroaryl group. X 1 If R is a sulfur atom or -S(=O)2-, 1 R represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 This represents an optionally substituted aryl group or an optionally substituted heteroaryl group. X 1 If R is an oxygen atom, 1 and R 2 Together with the sulfur and oxygen atoms to which they are bonded, they form a heterocycle which may be substituted, and X 1 -N(-R 3 )- If R 1 R represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 and R 3 They, together with the sulfur and nitrogen atoms to which they bond, form a heterocycle which may be substituted, or R 1 and R 3 Together with the nitrogen atoms to which they are bonded, they form a heterocycle which may be substituted, and R 2 This indicates an optionally substituted aryl group or an optionally substituted heteroaryl group. A method using a compound represented by (excluding 2,2-dipyridyl disulfide).

[0046]

[13] The compound represented by formula (I) is formula (Ia):

[0047] [ka]

[0048] [In the formula, R 1a This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Ring A exhibits a 5- or 6-membered unsaturated heterocycle. Y 1a This represents -S(=O)-, -S(=O)2-, -C(=O)-, or -C(=S)-. m represents an integer between 0 and 3. m R 2a Each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and If m is an integer greater than or equal to 2, then two adjacent R 2a It may form a fused ring, which may be substituted, together with ring A. Compounds represented by formula (Ib):

[0049] [ka]

[0050] [In the formula, R 1b This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Ring B represents a 6-membered aromatic hydrocarbon ring, or a 5- or 6-membered aromatic heterocycle. n represents an integer from 0 to 5. n R 2b Each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and If n is an integer greater than or equal to 2, then two adjacent R 2b It may form a substituted or bicyclic fused aromatic heterocycle together with ring B. Compounds represented by (excluding 2,2-dipyridyl disulfide) The method described in

[12] above.

[0051]

[14] The compound represented by formula (Ia) is, formula (Ic):

[0052] [ka]

[0053] [In the formula, R 1c This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Y 1c This represents -S(=O)-, -S(=O)2-, -C(=O)-, or -C(=S)-. p represents 0 or 1. R 2c This represents a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. Ring C represents a 6-membered aromatic hydrocarbon ring, a 6-membered nitrogen-containing aromatic heterocycle, or a 10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom. q represents an integer from 0 to 4, and q R 3c Each of these independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. The method according to

[13] , wherein the compound is represented by .

[0054]

[15] The compound represented by formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazole Lu-3(2H)-one, 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one, 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3 (2H)-one, 2-phenylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one, 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-phenyl-2H-1,2-benzothiadin-3(4H)-one, 2-(2,6-dimethylphenyl)isothiazolo[5,4-b The method according to

[14] , wherein at least one is selected from the group consisting of ]pyridine-3(2H)-one, 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one, 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one, and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one.

[0055]

[16] The method according to

[14] , wherein the ring C is a 6-membered aromatic hydrocarbon ring or a 6-membered nitrogen-containing aromatic heterocycle.

[17] The compound represented by formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazole-3(2H)-one, 2-(2,6-dimethylphenyl The method according to

[16] , wherein at least one is selected from the group consisting of phenyl)-5-nitro-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one, and 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3(2H)-one.

[0056]

[18] Formula (r1) in the compound represented by the above formula (Ib):

[0057] [ka]

[0058] The base represented by, (1) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (2) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (6) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (7) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (8) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (9) A phenyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (10) A 2-imidazolyl group which may be substituted with 1 to 3 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. The method described in any one of the above

[13] to

[17] .

[0059]

[19] The method according to

[18] , wherein the compound represented by formula (Ib) is at least one selected from the group consisting of 2,2'-dibenzothiazolyl disulfide, 2-(2-benzothiazolyldithio)propanoic acid, 3-(2-benzothiazolyldithio)propanoic acid, 2,2'-dibenzimidazolyl disulfide, 2,2'-dibenzoxazolyl disulfide, 5,5'-di(1,2,3-triazolyl) disulfide, 5,5'-dithiobis(1-phenyl-1H-tetrazole), 2,2'-dithiobis(pyridine-N-oxide), 4,4'-dipyridyl disulfide, 3-(2-pyridyldithio)propanoic acid, 2,2'-dithiobis(1H-imidazole), and 2,2'-dithiobis(1-methyl-1H-imidazole).

[0060]

[20] The method according to any one of the above

[12] to

[19] , wherein the quenching agent (i) is water.

[0061]

[21] The method according to any one of the above

[12] to

[20] , wherein the quenching agent (ii) is an organophosphorus compound.

[22] The method according to

[21] , wherein the organophosphorus compound is at least one selected from the group consisting of phosphines, phosphite triesters, phosphiniteroates, phosphoniteroates diesters and phosphiniteroates.

[23] The method according to

[21] , wherein the organophosphorus compound is a phosphine.

[0062]

[24] The method according to any one of the

[12] to

[23] , wherein the one-pot synthesis further comprises adding aromatic amines to the solution after step (3).

[0063]

[25] The method according to any one of the

[12] to

[24] , wherein the one-pot synthesis further comprises a step (6) of adding a base to the solution after step (5) and before step (7).

[0064]

[26] The method according to any one of

[12] to

[25] , wherein the oligonucleotide precursor (c) is an oligonucleotide precursor having a thiophosphate ester bond in addition to a phosphite ester bond.

[0065]

[27] Formula (In):

[0066] [ka]

[0067] [In the formula, (1)R 1n represents an optionally substituted phenyl group, p' represents 0, ring C'' represents a 10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom, and q' represents 0 or (2)R 1n represents an optional phenyl group, p' represents 0, ring C'' represents a 6-membered nitrogen-containing aromatic heterocycle, q' represents 1, and R 3n is C 1-6 Does it indicate a perfluoroalkyl group? (3)R 1n represents an optional phenyl group, p' represents 1, ring C'' represents a 6-membered nitrogen-containing aromatic heterocycle, and q' represents 0, or (4)R 1n is one C 1-6 This indicates a pyridyl group substituted with an alkyl group, where p' represents 0, the ring C'' represents a 6-membered nitrogen-containing aromatic heterocycle, and q' represents 0. A compound represented by the formula.

[0068]

[28] The compound according to

[27] , wherein the compound represented by formula (In) is 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one, 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-phenyl-2H-1,2-benzothiadin-3(4H)-one, or 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one. [Effects of the Invention]

[0069] According to the present invention, oligonucleotides can be produced while suppressing the formation of the aforementioned defective or desulfurized products compared to when iodine is used. [Modes for carrying out the invention]

[0070] 1. Terminology First, the terms used herein will be explained. Unless otherwise specified, the terms used herein have the same meaning as those generally understood by those skilled in the art to which this invention pertains.

[0071] 1-1. Terminology related to the base In this specification, "C a-b " means that the number of carbon atoms is between a and b (where a and b are integers). In this specification, "halogen atom" means a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom.

[0072] In this specification, "hydrocarbon group" refers to, for example, aliphatic hydrocarbon groups, aromatic aliphatic hydrocarbon groups, monocyclic saturated hydrocarbon groups, and aromatic hydrocarbon groups. Specific examples include monovalent and divalent groups such as alkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, aryl groups, aralkyl groups, and alkylene groups.

[0073] In this specification, the alkyl(group) may be linear or branched. Examples of the alkyl(group) include alkyl groups having 1 or more carbon atoms, and when there is no particular limitation on the carbon number range, C is preferred. 1-10 It is an alkyl group, more preferably C 1-6 These are alkyl groups. Specific examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl.

[0074] In this specification, the "alkenyl (group)" may be either linear or branched. Examples of the "alkenyl (group)" include C 2-6 alkenyl groups and the like. Specifically, for example, vinyl, 1-propenyl, allyl, isopropenyl, butenyl, isobutenyl and the like can be mentioned.

[0075] In this specification, the "alkynyl (group)" may be either linear or branched. Examples of the "alkynyl (group)" include C 2-6 alkynyl groups and the like. Specific examples thereof include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 5-hexynyl and the like.

[0076] In this specification, the "cycloalkyl (group)" means a cyclic alkyl group, and examples thereof include C 3-8 cycloalkyl groups, preferably C 3-6 cycloalkyl groups. Specific examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.

[0077] In this specification, the "aryl (group)" means a monocyclic or polycyclic (condensed) aromatic hydrocarbon group, and examples thereof include C 6-14 aryl groups such as phenyl, 1-naphthyl, 2-naphthyl, biphenylyl, 1-anthryl, 2-anthryl and the like. Among them, C 6-10 aryl groups are more preferred, and phenyl is particularly preferred.

[0078] In this specification, the "heteroaryl (group)" means a monocyclic or polycyclic (condensed) aromatic heterocyclic group containing, as ring-constituting atoms, heteroatoms selected from nitrogen atoms, sulfur atoms and oxygen atoms in addition to carbon atoms. Examples of the heteroaryl group include 5- or 6-membered monocyclic heteroaryl groups and 8- to 14-membered condensed polycyclic heteroaryl groups.

[0079] In this specification, "5 or 6-membered monocyclic heteroaryl (group)" includes, for example, thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, (pyridine-N-oxide)-yl, pyrazinyl, pyrimidinyl, pyridadinyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and the like. In the present invention, "heteroaryl group" includes the (pyridine-N-oxide)-yl group represented by the following formula (wherein * indicates the bond position).

[0080] [ka]

[0081] In this specification, "5- or 6-membered monocyclic nitrogen-containing heteroaryl (group)" refers, for example, to the aforementioned "5- or 6-membered monocyclic heteroaryl (group)" that contains at least one nitrogen atom as a ring constituent atom.

[0082] In this specification, examples of the "8- to 14-member fused polycyclic heteroaryl (group)" include benzothiophenyl, benzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, benzotriazolyl, imidazopyridinyl, thienopyridinyl, furopyridinyl, pyrrolopyridinyl, pyrazolopyridinyl, oxazolopyridinyl, thiazolopyridinyl, imidazopyrazinyl, imidazopyrimidinyl, thienopyrimidinyl, furopyrimidinyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl, oxazolopyrimidinyl, thiazolopyrimidinyl, pyrazolotriazinyl, naphtho[2,3-b]thienyl, phenoxathiinyl, indolyl, isoindolyl, 1H-indazolyl, purinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, and the like.

[0083] In this specification, examples of the "8- to 14-member fused polycyclic nitrogen-containing heteroaryl (group)" include those among the aforementioned "8- to 14-member fused polycyclic heteroaryl (group)" that contain at least one or more nitrogen atoms as ring-constituting atoms.

[0084] In this specification, the "aralkyl (group)" includes a 7-20 aralkyl group, preferably a 7-16 aralkyl group ( 6-10 aryl-C 1-6 alkyl group). Specific examples thereof include benzyl, 1-phenylethyl, 2-phenylethyl, 1-phenylpropyl, naphthylmethyl, 1-naphthylethyl, 1-naphthylpropyl, and the like.

[0085] In this specification, the "alkylene (group)" may be either linear or branched. Examples of the "alkylene (group)" include alkylene groups having 1 or more carbon atoms. When there is no limitation on the carbon number range, it is preferably a 1-10 alkylene group, more preferably a1-6 These are alkylene groups. Specific examples include methylene, ethylene, propylene, butylene, pentylene, and hexylene.

[0086] In this specification, the "alkoxy(group)" may be linear or branched. Examples of the "alkoxy(group)" include alkoxy groups having 1 or more carbon atoms, and when there is no particular limitation on the carbon number range, C is preferred. 1-10 It is an alkoxy group, more preferably C 1-6 These are alkoxy groups. Specific examples include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, and hexyloxy.

[0087] In this specification, the "acyl(group)" may be linear or branched. Examples of the "acyl(group)" include C 1-6 Alkanoyl group, C 7-13 Examples include the aroyl group. Specific examples of "acyl(group)" include formyl, acetyl, n-propionyl, isopropionyl, n-butyryl, isobutyryl, pivaloyl, valeryl, hexanoyl, benzoyl, naphthoyl, and levlinyl. The "acyl(group)" may be substituted.

[0088] In this specification, “six-membered aromatic hydrocarbon ring” means a benzene ring. If the “six-membered aromatic hydrocarbon ring” forms part of a fused ring, then “six-membered aromatic hydrocarbon ring” means a benzene ring that forms part of a fused ring.

[0089] In this specification, "5 or 6-membered aromatic heterocycles" include, for example, thiophene rings, furan rings, 1H-pyrrole rings, imidazole rings, pyrazole rings, thiazole rings, isothiazole rings, oxazole rings, isoxazole rings, 1,2,4-oxadiazole rings, 1,3,4-oxadiazole rings, 1,2,4-thiadiazole rings, 1,3,4-thiadiazole rings, triazole rings, tetrazole rings, pyridine rings, pyridine-N-oxide rings, pyrazine rings, pyrimidine rings, pyridazine rings, triazine rings, and the like. In this invention, "aromatic heterocycle" includes pyridine-N-oxide rings.

[0090] In this specification, "4- to 8-membered nitrogen-containing heterocycles" include, for example, azetidine rings, pyrrolidine rings, pyrroline rings, 2H-pyrrole rings, piperidine rings, piperazine rings, azacycloheptane rings, azacyclooctane rings, 1H-pyrrole rings, imidazole rings, pyrazole rings, thiazole rings, isothiazole rings, oxazole rings, isoxazole rings, 1,2,4-oxadiazole rings, 1,3,4-oxadiazole rings, 1,2,4-thiadiazole rings, 1,3,4-thiadiazole rings, triazole rings, tetrazole rings, pyridine rings, pyridine-N-oxide rings, pyrazine rings, pyrimidine rings, pyridazine rings, triazine rings, and the like.

[0091] In this specification, "5- or 6-membered nitrogen-containing heterocycles" include, for example, those among the aforementioned "4- to 8-membered nitrogen-containing heterocycles" that have 5 or 6 ring-constituting atoms.

[0092] In this specification, "six-membered nitrogen-containing aromatic heterocycle" refers to, for example, a pyridine ring, a pyridine-N-oxide ring, a pyrazine ring, a pyrimidine ring, a pyridazine ring, a triazine ring, and the like.

[0093] In this specification, "bicyclic condensed nitrogen-containing heterocycles" include, for example, pyrrolidine rings, decahydroisoquinoline rings, decahydroquinoline rings, benzothiophene rings, benzofuran rings, isobenzofuran rings, benzimidazole rings, benzoxazole rings, benzoisoxazole rings, benzothiazole rings, benzoisothiazole rings, benzotriazole rings, indole rings, isoindole rings, 1H-indazole rings, purine rings, isoquinoline rings, 5,6,7,8-tetrahydroisoquinoline rings, quinoline rings, 5,6,7,8-tetrahydroquinoline rings, phthalazine rings, pteridine rings, naphthyridine rings, quinoxaline rings, quinazoline rings, sinnoline rings, and the like.

[0094] In this specification, "bicyclic fused aromatic heterocycles" include, for example, benzothiophene rings, benzofuran rings, isobenzofuran rings, benzimidazole rings, benzoxazole rings, benzoisoxazole rings, benzothiazole rings, benzoisothiazole rings, benzotriazole rings, indole rings, isoindole rings, 1H-indazole rings, purine rings, isoquinoline rings, 5,6,7,8-tetrahydroisoquinoline rings, quinoline rings, 5,6,7,8-tetrahydroquinoline rings, phthalazine rings, pteridine rings, naphthyridine rings, quinoxaline rings, quinazoline rings, sinnoline rings, and the like. In this specification, "bicyclic fused aromatic heterocycles" also include bicyclic fused rings in which one ring is an aromatic heterocycle and the other ring is a non-aromatic heterocycle or non-aromatic hydrocarbon ring (for example, a 5,6,7,8-tetrahydroisoquinoline ring).

[0095] In this specification, "10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom" refers to, for example, a 5,6,7,8-tetrahydroisoquinoline ring, a quinoline ring, and the like.

[0096] In this specification, examples of "electron-withdrawing groups" include halogen atoms, perchloroalkyl groups (e.g., trichloromethyl group), perfluoroalkyl groups (e.g., trifluoromethyl group), nitro groups, cyano groups, formyl groups, alkylcarbonyl groups (-CO-R, R: alkyl group), perchloroalkylcarbonyl groups (-CO-R, R: perchloroalkyl group), perfluoroalkylcarbonyl groups (-CO-R, R: perfluoroalkyl group), carboxyl groups, alkoxycarbonyl groups (-CO-OR, R: alkyl group), alkylsulfonyl groups (-S(=O)2-R, R: alkyl group), sulfo groups, and the like.

[0097] In this specification, "perchloroalkyl group" means an alkyl group in which all hydrogen atoms are replaced by chlorine atoms. In this specification, "perfluoroalkyl group" means an alkyl group in which all hydrogen atoms are replaced by fluorine atoms.

[0098] In this specification, "linker" means, for example, -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -NR'-, -C(=O)NR'-, -NR'C(=O)-, -S-, -SO-, -SO2-, -Si(R')(R”)O-, -Si(R')(R”)-(R' and R'' are each independently a hydrogen atom or C 1-22 Examples include hydrocarbon groups. The linker is preferably -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH-, -NHC(=O)-, -S-, -SO-, -SO2-, -Si(R')(R”)O-, or -Si(R')(R”)-(R' and R'' are each independently hydrogen atoms or C 1-22 (This indicates a hydrocarbon group.)

[0099] In this specification, "substituents" include, for example, the halogen atoms, alkyl groups, aralkyl groups, alkoxy groups, acyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, and aryl groups mentioned above, as well as hydroxyl groups, nitro groups, cyano groups, guanidyl groups, carboxyl groups, alkoxycarbonyl groups (the alkoxy portion is the same as that of the alkoxy group), sulfo groups, phospho groups, alkylsulfanyl groups (the alkyl portion is the same as that of the alkyl group), alkylsulfinyl groups (the alkyl portion is the same as that of the alkyl group), alkylsulfonyl groups (the alkyl portion is the same as that of the alkyl group), amino groups, monoalkylamino groups (the alkyl portion is the same as that of the alkyl group), dialkylamino groups (the alkyl portion is the same as that of the alkyl group), oxo groups, and the like.

[0100] In this specification, there are no particular limitations on the "hydroxyl group protecting group," and known protecting groups can be used. Examples of such protecting groups include methyl group, benzyl group, p-methoxybenzyl group, tert-butyl group, methoxymethyl group, 2-methoxyethyl group, 2-tetrahydropyranyl group, 2-ethoxyethyl group, 2-cyanoethyl group, (2-cyanoethoxy)methyl group, 1-(2-cyanoethoxy)ethyl group, bis(2-acetoxyethoxy)methyl (ACE) group, (2-nitrobenzyl)oxymethyl (NBOM) group, (2-(trimethylsilyl)ethoxy)methyl (SEM) group, 1-(2-cyanoethoxy)ethyl group, 2-((4-methylphenyl)sulfonyl)ethoxymethyl (TEM) group, tert-butyldithiomethyl (DTM) group, ((2-(methylthio)phenyl Examples include the thio)methyl (MPTM) group, the (N-dichloroacetyl-N-methyl)aminobenzyloxymethyl (DCMABOM) group, the (2-acetylethyl)carbonyloxymethyl group (also known in this field as the "acetallevnyl ester (ALE) group"), the phenylcarbamoyl group, the 1,1-dioxothiomorpholine-4-thiocarbamoyl group, the acetyl group, the pivaloyl group, the benzoyl group, the trimethylsilyl group, the triethylsilyl group, the triisopropylsilyl group, the tert-butyldimethylsilyl group, the [(triisopropylsilyl)oxy]methyl (Tom) group, and the 1-(4-chlorophenyl)-4-ethoxypiperidine-4-yl (Cpep) group.

[0101] In this specification, "temporary protecting groups that can be removed under acidic conditions of a hydroxyl group" include, for example, trityl group, 9-phenyl-9-xanthenyl group, 9-phenyl-9-thioxanthenyl group, 1,1-bis(4-methoxyphenyl)-1-phenylmethyl group (sometimes referred to as "4,4'-dimethoxytrityl group" in this specification), and other bis(C) groups. 1-6 Mono(C) groups such as alkoxy)trityl group, 1-(4-methoxyphenyl)-1,1-diphenylmethyl group (sometimes referred to as "4-monomethoxytrityl group" in this specification) 1-18Examples include alkoxy)trityl groups. From the viewpoint of ease of deprotection, the primary protecting group is preferably a 4,4'-dimethoxytrityl group or a 4-monomethoxytrityl group, and more preferably a 4,4'-dimethoxytrityl group.

[0102] In this specification, there are no particular limitations on the "amino group protecting group," and known protecting groups can be used. Examples of such protecting groups include the pivaloyl group, pivaloyloxymethyl group, acetyl group, trifluoroacetyl group, phenoxyacetyl group, 4-isopropylphenoxyacetyl group, 4-tert-butylphenoxyacetyl group, benzoyl group, isobutyryl group, (2-hexyl)decanoyl group, dimethylformamidinyl group, 1-(dimethylamino)ethylidene group, and 9-fluorenylmethyloxycarbonyl group. Among these, acetyl group, phenoxyacetyl group, 4-isopropylphenoxyacetyl group, benzoyl group, isobutyryl group, (2-hexyl)decanoyl group, dimethylformamidinyl group, and =NC(R 11 )-N(R 12 )(R 13 A base represented by (wherein R in the formula) 11 R indicates a methyl group. 12 and R 13 Each of them is independently C 1-6 Indicates an alkyl group, or R 11 and R 12 These may come together to form a 5- or 6-membered nitrogen-containing heterocycle with the carbon and nitrogen atoms to which they are bonded. ) are examples. The above = NC(R 11 )-N(R 12 )(R 13 An example of a group represented by ) is the 1-(dimethylamino)ethylidene group.

[0103] In this specification, there are no particular limitations on the term "phosphate protecting group," and any known protecting group may be used. Examples of phosphate protecting groups include: The 2-cyanoethyl group represented by the following formula (Pg-1), The 2-[2-(4,4'-dimethoxytrityloxy)ethylsulfonyl]ethyl group represented by the following formula (Pg-2), The [3-(4,4'-dimethoxytrityloxy)-2,2-di(ethoxycarbonyl)]propyl group, represented by the following formula (Pg-3), The [3-(4,4'-dimethoxytrityloxy)-2,2-di(N-methylcarbamoyl)]propyl group is represented by the following formula (Pg-4). The protecting group for the phosphate group is preferably a 2-cyanoethyl group.

[0104] [ka]

[0105] [In the formula, Me represents a methyl group, Et represents an ethyl group, DMTr represents a 4,4'-dimethoxytrityl group, and * indicates the bond position to the phosphate group.]

[0106] In this specification, as a "hydrophobic protecting group," for example, a protecting group described as "anchor" in International Publication No. 2017 / 104836 may be used.

[0107] The hydrophobic protecting group is preferably a protecting group represented by the following formula (Pg-5): **LYZ (Pg-5) [In the formula, ** indicates the bond position with the protected group; L is a single bond, or formula (Pg-i-1) or (Pg-i-2):

[0108] [ka]

[0109] (In the formula, * indicates the bond position with Y; ** is synonymous with the above; R 1p and R 2p Each of them is independent of C1-22 Showing hydrocarbon groups; L 1 This is a divalent C which may be substituted. 1-22 It represents a hydrocarbon group, and the aforementioned divalent C 1-22 The -CH2- group in the hydrocarbon group may be replaced by a linker; L 2 This indicates a single bond, or ***C(=O)N(R 3p )-R 4p -N(R 5p )****(In the formula, *** is L 1 The bond position is shown, **** indicates the bond position with C=O, and R 4p C 1-22 It shows an alkylene group, R 3p and R 5p Each of these is independently a hydrogen atom or C 1-22 Indicates an alkyl group, or R 3p and R 5p They may come together to form a ring. (This indicates a group represented by ). (This indicates a group represented by ); Y represents a single bond, an oxygen atom, or NR (where R represents a hydrogen atom, an alkyl group, or an aralkyl group); and Z is one of the following equations: (Pg-ii-1) to (Pg-ii-5):

[0110] [ka]

[0111] [In the formula, * indicates the bond location; Ring A' represents a benzene ring; Ring B' represents a cyclohexane ring; Ring D' represents a naphthalene ring or a bicyclic fused aromatic heterocycle; R 6p is either a hydrogen atom or R b However, if the group is represented by the following formula (Pg-iii), then the R of ring A' or ring B' 6p R 8pThey may also form a single bond or an -O- in conjunction with ring A' or ring B' and ring C', forming a fused ring; k represents an integer between 1 and 4; k Qs are each independently -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -NR'-, -C(=O)NR'-, or -NR'C(=O)- (wherein R' is each independently a hydrogen atom or C 1-6 (representing an alkyl group) (where k Qs preferably each independently represent -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-); k Q' are each independently a single bond, -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -NR'-, -C(=O)NR'-, or -NR'C(=O)- (wherein R' is each independently a hydrogen atom or C 1-6 (Indicates an alkyl group) k pieces R 7p Each of these independently represents a hydrocarbon group in which a linear aliphatic hydrocarbon group having 10 or more carbon atoms is bonded either by a single bond or via a linker; Ring A' and Ring B' each independently contain k QR codes. 7p In addition, C may be substituted with halogen atoms or halogen atoms. 1-6 C which may be substituted with alkyl and halogen atoms 1-6 They may also have substituents selected from the group consisting of alkoxy groups; R a This indicates a hydrogen atom; and R b is a hydrogen atom, or formula (Pg-iii):

[0112] [ka]

[0113] (In the formula, * indicates the bond location; Ring C' represents a benzene ring: j represents an integer between 0 and 4; Each of the j Qs is independently equivalent to the above; j R 9p Each of these independently represents a hydrocarbon group in which a linear aliphatic hydrocarbon group having 10 or more carbon atoms is bonded either by a single bond or via a linker; R 8p This indicates a hydrogen atom, or the R of ring A' or ring B'. 6p They may also form a single bond or an -O- in conjunction with ring A' or ring B' and ring C', forming a fused ring; Ring C' contains j QRs. 9p In addition, C may be substituted with halogen atoms or halogen atoms. 1-6 C which may be substituted with alkyl and halogen atoms 1-6 It may have substituents selected from the group consisting of alkoxy groups.) or it may represent a group represented by ) R a and R b However, together they form an oxo group; u indicates 1 or 2; and R 10p C 2-21 This indicates a perfluoroalkyl group. This indicates the group represented by [ ].

[0114] Note that rings A', C', and R 6p and R 8p When a fused ring is formed, ring A' and ring C' each represent the benzene ring portion of the fused ring. Ring B', ring C', R 6p and R 8p When a fused ring is formed, ring C' represents the benzene ring portion of the fused ring, and ring B' represents the cyclohexane ring portion of the fused ring.

[0115] The linear aliphatic hydrocarbon group having 10 or more carbon atoms is preferably a linear C 10-40 Alkyl and linear C 10-40 A group selected from alkenyl groups, more preferably a linear C group. 10-40 It is an alkyl group, and more preferably a linear C 10-30It is an alkyl group, and particularly preferably a linear C 12-28 It is an alkyl group, most preferably a linear C 14-26 It is an alkyl group.

[0116] The linker is preferably -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -NR'-, -C(=O)NR'-, or -NR'C(=O)-(R' is a hydrogen atom or C 1-22 It represents a hydrocarbon group, more preferably -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-, and even more preferably -O-.

[0117] The aforementioned "hydrocarbon group in which a linear aliphatic hydrocarbon group having 10 or more carbon atoms is bonded via a single bond or a linker" is preferably a linear C 10-40 Alkyl group, 1-3 linear carbon atoms 10-40 A benzyl group to which an alkyl group is bonded via -O-, or 1 to 3 linear carbon atoms. 10-40 This is a cyclohexylmethyl group to which an alkyl group is bonded via an -O- junction.

[0118] Q is preferably -O-, -C(=O)NH-, or -NHC(=O)-, and more preferably -O-.

[0119] Q' is preferably a single bond, -O-, -C(=O)NH- or -NHC(=O)-, and more preferably a single bond or -O-.

[0120] In formula (Pg-5), a preferred embodiment of L represented by formula (Pg-i-1) is: L 1 However, divalent C 1-22 A hydrocarbon group, or CH2-O-1,4-phenylene-O-CH2; and L 2 However, it is either a single bond or ***C(=O)N(R 3p )-R 4p -N(R 5p )****(In the formula, *** is L 1The bond position is shown, **** indicates the bond position with C=O, and R 4p C 1-6 It shows an alkylene group, R 3p and R 5p Each of these is independently a hydrogen atom, or a substituted C atom. 1-6 Indicates an alkyl group, or R 3p and R 5p These can be combined and replaced with C 1-6 It may form an alkylene group. It is the basis.

[0121] Another preferred embodiment of L represented by formula (Pg-i-1) is, L 1 However, divalent C 1-22 It is a hydrocarbon group; and L 2 is a single bond It is the basis.

[0122] Another preferred embodiment of L represented by formula (Pg-i-1) is, L 1 However, it is an ethylene group; and L 2 However, ***C(=O)N(R 3p )-R 4p -N(R 5p )****(In the formula, *** is L 1 The bond position is shown, **** indicates the bond position with C=O, and R 4p C 1-22 It shows an alkylene group, R 3p and R 5p Each of these is independently a hydrogen atom or C 1-22 Indicates an alkyl group, or R 3p and R 5p They may come together to form a ring. ) is a group represented by It is the basis.

[0123] Another preferred embodiment of L represented by formula (Pg-i-1) is, L 1 However, it is an ethylene group; and L 2 However, ***C(=O)N(R 3p )-R 4p -N(R 5p )****(In the formula, *** is L 1 The bond position is indicated, and **** indicates the bond position with C=O, and N(R 3p )-R 4p -N(R 5p The portion in question forms a 1,4-piperazinediyl group. It is the basis.

[0124] Another preferred embodiment of L represented by formula (Pg-i-1) is, L 1 However, it is an ethylene group; and L 2 However, ***C(=O)N(R 3p )-R 4p -N(R 5p )****(In the formula, *** is L 1 The bond position is shown, **** indicates the bond position with C=O, and R 4p R represents a pentylene group or a hexylene group. 3p and R 5p Each of these independently represents either a hydrogen atom or a methyl group. It is the basis.

[0125] Another preferred embodiment of L represented by formula (Pg-i-1) is, L 1 However, it is a phenylene group which may have substituents; and L 2 However, ***C(=O)N(R 3p )-R 4p -N(R 5p )****(In the formula, *** is L 1 The bond position is indicated, and **** indicates the bond position with C=O, and N(R 3p )-R 4p -N(R 5p The portion in question forms a 1,4-piperazinediyl group. It is the basis.

[0126] In formula (Pg-i-1), L is particularly preferably a succinyl group.

[0127] Next, we will explain L, which is expressed in equation (Pg-i-2) in equation (Pg-5). In equation (Pg-i-2), L 1 Preferably, divalent C 6-10 It is an aromatic hydrocarbon group, and more preferably a phenylene group. L in equation (Pg-i-2) 2 The bond is preferably a single bond. In equation (Pg-i-2), L 1 and L 2 A preferred combination is L 1 is a divalent C 6-10 It is an aromatic hydrocarbon group, L 2 It is a single bond, a combination. In equation (Pg-i-2), L 1 and L 2 A more preferred combination is L 1 The phenylene group is L 2 This is a combination of single bonds. R in equation (Pg-i-2) 1p and R 2p Each of these is independently, preferably C 1-22 It is an alkyl group, more preferably C 1-10 It is an alkyl group.

[0128] A preferred embodiment of L represented by formula (Pg-i-2) is: R 1p and R 2p However, each independently, C 1-22 It is an alkyl group; L 1 However, divalent C 6-10 It is an aromatic hydrocarbon group; and L 2 However, it is a single bond. It is the basis.

[0129] Another preferred embodiment of L represented by formula (Pg-i-2) is, R 1p and R 2pHowever, each independently, C 1-10 It is an alkyl group; L 1 However, it is a phenylene group; and L 2 However, it is a single bond. It is the basis.

[0130] When Y in formula (Pg-5) is NR, R is preferably a hydrogen atom, C 1-6 Alkyl or C 7-16 The group is an aralkyl group, more preferably a hydrogen atom, methyl, ethyl, or benzyl, and even more preferably a hydrogen atom. Y is preferably a single bond, an oxygen atom, or NR, and more preferably a single bond or an oxygen atom.

[0131] In equation (Pg-ii-1), R 6p It is preferable that is a hydrogen atom. Also, in formula (Pg-ii-1), R a and R b However, it is preferable that it be a hydrogen atom or, together with it, form an oxo group.

[0132] A preferred embodiment of Z, represented by formula (Pg-ii-1), is R a and R b However, it is a hydrogen atom; R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each is independent of the linear C 10-40 It is an alkyl group. It is the basis.

[0133] Another preferred embodiment of Z, represented by formula (Pg-ii-1), is R a and R b However, it is a hydrogen atom; R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each independently consists of 1 to 3 linear C atoms. 10-40 A benzyl group to which an alkyl group is bonded via -O-, or 1 to 3 linear carbon atoms. 10-40 It is a cyclohexylmethyl group to which an alkyl group is bonded via an -O-. It is the basis.

[0134] Another preferred embodiment of Z, represented by formula (Pg-ii-1), is R a However, it is a hydrogen atom; R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each is independent of the linear C 10-40 It is an alkyl group; and R b However, equation (Pg-iii) (where * is the joining position, j is an integer from 0 to 3, j Qs are -O-, and j Rs) 9p However, each is independent of the linear C 10-40 It is an alkyl group, and R 8p However, it is a hydrogen atom.) It is the basis.

[0135] Another preferred embodiment of Z, represented by formula (Pg-ii-1), is R a However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each is independent of the linear C 10-40 It is an alkyl group; and R b However, equation (Pg-iii) (where * is the joining position, j is an integer from 0 to 3, j Qs are -O-, and j Rs) 9p However, each is independent of the linear C 10-40 It is an alkyl group, and R8p R 6p Together with it, it shows a single bond or -O- and forms a fused ring with ring A' and ring C'. ) This is the group represented by It is the basis.

[0136] Another preferred embodiment of Z, represented by formula (Pg-ii-1), is R a and R b However, together they form an oxo group; R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each is independent of the linear C 10-40 It is an alkyl group. It is the basis.

[0137] Another preferred embodiment of Z, represented by formula (Pg-ii-1), is R a and R b However, together they form an oxo group; R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; k pieces R 7p However, each independently consists of 1 to 3 linear C atoms. 10-40 A benzyl group to which an alkyl group is bonded via -O-, or 1 to 3 linear carbon atoms. 10-40 It is a cyclohexylmethyl group to which an alkyl group is bonded via an -O-. It is the basis.

[0138] A preferred embodiment of Z, represented by formula (Pg-ii-2), is R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each is independent of the linear C 10-40Alkyl group, 1-3 linear carbon atoms 10-40 A benzyl group to which an alkyl group is bonded via -O-, or 1 to 3 linear carbon atoms. 10-40 It is a cyclohexylmethyl group to which an alkyl group is bonded via an -O-. It is the basis.

[0139] In equation (Pg-ii-3), R 6p It is preferable that is a hydrogen atom. Also, in formula (Pg-ii-3), R a and R b However, it is preferable that it be a hydrogen atom or, together with it, form an oxo group.

[0140] A preferred embodiment of Z, represented by formula (Pg-ii-3), is R a and R b However, it is a hydrogen atom; R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each is independent of the linear C 10-40 It is an alkyl group. It is the basis.

[0141] Another preferred embodiment of Z, represented by formula (Pg-ii-3), is R a and R b However, it is a hydrogen atom; R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each independently consists of 1 to 3 linear C atoms. 10-40 A benzyl group to which an alkyl group is bonded via -O-, or 1 to 3 linear carbon atoms. 10-40 It is a cyclohexylmethyl group to which an alkyl group is bonded via an -O-. It is the basis.

[0142] Another preferred embodiment of Z, represented by formula (Pg-ii-3), is R a However, it is a hydrogen atom; R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each is independent of the linear C 10-40 It is an alkyl group; and R b However, equation (Pg-iii) (where * is the joining position, j is an integer from 0 to 3, j Qs are -O-, and j Rs) 9p However, each is independent of the linear C 10-40 It is an alkyl group, and R 8p However, it is a hydrogen atom.) It is the basis.

[0143] Another preferred embodiment of Z, represented by formula (Pg-ii-3), is R a However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each is independent of the linear C 10-40 It is an alkyl group; and R b However, equation (Pg-iii) (where * is the joining position, j is an integer from 0 to 3, j Qs are -O-, and j Rs) 9p However, each is independent of the linear C 10-40 It is an alkyl group, and R 8p R 6p Together with it, it shows a single bond or -O- and forms a fused ring with ring B' and ring C'. ) This is the group represented by It is the basis.

[0144] Another preferred embodiment of Z, represented by formula (Pg-ii-3), is R a and R bHowever, together they form an oxo group; R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; and k pieces R 7p However, each is independent of the linear C 10-40 It is an alkyl group. It is the basis.

[0145] Another preferred embodiment of Z, represented by formula (Pg-ii-3), is R a and R b However, together they form an oxo group; R 6p However, it is a hydrogen atom; k is an integer between 1 and 3; k Qs are -O-; k pieces R 7p However, each independently consists of 1 to 3 linear C atoms. 10-40 A benzyl group to which an alkyl group is bonded via -O-, or 1 to 3 linear carbon atoms. 10-40 It is a cyclohexylmethyl group to which an alkyl group is bonded via an -O-. It is the basis.

[0146] A preferred embodiment of Z, represented by formula (Pg-ii-4), is Ring D' is a naphthalene ring; R a and R b However, it is a hydrogen atom; k is an integer between 1 and 3; k Q' are -O- and k pieces R 7p However, each is independent of the linear C 10-40 Alkyl group, 1-3 linear carbon atoms 10-40 A benzyl group to which an alkyl group is bonded via -O-, or 1 to 3 linear carbon atoms. 10-40 It is a cyclohexylmethyl group to which an alkyl group is bonded via an -O-. It is the basis.

[0147] Another preferred embodiment of Z, represented by formula (Pg-ii-4), is Ring D' is an indole ring; R a and R b However, it is a hydrogen atom; k is 1; Q' is a single bond; and R 7p However, linear C 10-40 Alkyl group, 1-3 linear carbon atoms 10-40 A benzyl group to which an alkyl group is bonded via -O-, or 1 to 3 linear carbon atoms. 10-40 A cyclohexylmethyl group to which an alkyl group is bonded via -O-; and R 7p However, it is bonded to the nitrogen atom of the indole ring. It is the basis.

[0148] A preferred embodiment of Z, represented by formula (Pg-ii-5), is u is 1 or 2; and R 10p However, C 4-10 It is a perfluoroalkyl group. It is the basis.

[0149] Z is preferably a group represented by formula (Pg-ii-1), formula (Pg-ii-2), or formula (Pg-ii-3).

[0150] The protecting group (Pg-5) is preferably, L is either a succinyl group or a group represented by formula (Pg-i-2) (where R is...). 1p and R 2p However, each independently, C 1-10 It is an alkyl group, L 1 However, it is a divalent phenylene group, L 2 However, it is a single bond. ) and YZ is a 3,4,5-tris(octadecyloxy)benzyloxy group, a 3,5-bis(docosyloxy)benzyloxy group, a 3,5-bis[3',4',5'-tris(octadecyloxy)benzyloxy]benzyloxy group, a 3,4,5-tris[3',4',5'-tris(octadecyloxy)benzyloxy]benzyloxy group, a 3,4,5-tris(octadecyloxy)benzylamino group, a 2,4-bis(docosyloxy)benzylamino group, a 3,5-bis(docosyloxy)benzylamino group, and a bis(4-docosyloxyphenyl) group. Methylamino group, 4-methoxy-2-[3',4',5'-tris(octadecyloxy)benzyloxy]benzylamino group, 4-methoxy-2-[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]benzylamino group, 2,4-bis(dodecyloxy)benzylamino group, phenyl(2,3,4-tris(octadecyloxy)phenyl)methylamino group, bis[4-(12-docosyloxydodecyloxy)phenyl]methylamino group, 3,5-bis[3',4',5'-tris(octadecyloxy)benzyloxy [Xy]benzylamino group, 3,4,5-tris[3',4',5'-tris(octadecyloxy)benzyloxy]benzylamino group, 3,4,5-tris(octadecyloxy)cyclohexylmethyloxy group, 3,5-bis(docosyloxy)cyclohexylmethyloxy group, 3,5-bis[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethyloxy group, 3,4,5-tris[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethyloxy group, 3,4,5-Tris(octadecyloxy)cyclohexylmethylamino group, 2,4-Bis(docosyloxy)cyclohexylmethylamino group, 3,5-Bis(docosyloxy)cyclohexylmethylamino group, Bis(4-docosyloxycyclohexyl)methylamino group, 4-Methoxy-2-[3',4',5'-Tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group, 4-Methoxy-2-[3',4',5'-Tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group, 2,The group is a 4-bis(dodecyloxy)cyclohexylmethylamino group, a phenyl(2,3,4-tris(octadecyloxy)cyclohexyl)methylamino group, a bis[4-(12-docosyloxydodecyloxy)cyclohexyl]methylamino group, a 3,5-bis[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group, or a 3,4,5-tris[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group. is a base, or LY is a single bond or succinyl-1,4-piperazinediyl group, and Z is a 3,4,5-tris(octadecyloxy)benzoyl group, a 3,5-bis(docosyloxy)benzoyl group, a 3,5-bis[3',4',5'-tris(octadecyloxy)benzyloxy]benzoyl group, or a 3,4,5-tris[3',4',5'-tris(octadecyloxy)benzyloxy]benzoyl group. It is the basis.

[0151] The protecting group (Pg-5) is more preferably, L is a succinyl group, and YZ is a 3,4,5-tris(octadecyloxy)benzyloxy group, a 3,5-bis(docosyloxy)benzyloxy group, a 3,5-bis[3',4',5'-tris(octadecyloxy)benzyloxy]benzyloxy group, a 3,4,5-tris[3',4',5'-tris(octadecyloxy)benzyloxy]benzyloxy group, a 3,4,5-tris(octadecyloxy)benzylamino group, a 2,4-bis(docosyloxy)benzylamino group, a 3,5-bis(docosyloxy)benzylamino group, and a bis(4-docosyloxyphenyl) group. Methylamino group, 4-methoxy-2-[3',4',5'-tris(octadecyloxy)benzyloxy]benzylamino group, 4-methoxy-2-[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]benzylamino group, 2,4-bis(dodecyloxy)benzylamino group, phenyl(2,3,4-tris(octadecyloxy)phenyl)methylamino group, bis[4-(12-docosyloxydodecyloxy)phenyl]methylamino group, 3,5-bis[3',4',5'-tris(octadecyloxy)benzyloxy [Xy]benzylamino group, 3,4,5-tris[3',4',5'-tris(octadecyloxy)benzyloxy]benzylamino group, 3,4,5-tris(octadecyloxy)cyclohexylmethyloxy group, 3,5-bis(docosyloxy)cyclohexylmethyloxy group, 3,5-bis[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethyloxy group, 3,4,5-tris[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethyloxy group, 3,4,5-Tris(octadecyloxy)cyclohexylmethylamino group, 2,4-Bis(docosyloxy)cyclohexylmethylamino group, 3,5-Bis(docosyloxy)cyclohexylmethylamino group, Bis(4-docosyloxycyclohexyl)methylamino group, 4-Methoxy-2-[3',4',5'-Tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group, 4-Methoxy-2-[3',4',5'-Tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group, 2,The group is a 4-bis(dodecyloxy)cyclohexylmethylamino group, a phenyl(2,3,4-tris(octadecyloxy)cyclohexyl)methylamino group, a bis[4-(12-docosyloxydodecyloxy)cyclohexyl]methylamino group, a 3,5-bis[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group, or a 3,4,5-tris[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group. is a base, or LY is a single bond or succinyl-1,4-piperazinediyl group, and Z is a 3,4,5-tris(octadecyloxy)benzoyl group, a 3,5-bis(docosyloxy)benzoyl group, a 3,5-bis[3',4',5'-tris(octadecyloxy)benzyloxy]benzoyl group, or a 3,4,5-tris[3',4',5'-tris(octadecyloxy)benzyloxy]benzoyl group. It is the basis.

[0152] The protecting group (Pg-5) is more preferably, L is a succinyl group, and YZ is a 3,4,5-tris(octadecyloxy)benzyloxy group, a 3,4,5-tris(octadecyloxy)cyclohexylmethyloxy group, a 3,5-bis(docosyloxy)cyclohexylmethyloxy group, a 3,5-bis[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethyloxy group, a 3,4,5-tris[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethyloxy group, a 3,4,5-tris(octadecyloxy)cyclohexylmethylamino group, a 2,4-bis(docosyloxy)cyclohexylmethylamino group, a 3,5-bis(docosyloxy The group is a cyclohexylmethylamino group, a 4-methoxy-2-[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group, a 4-methoxy-2-[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group, a 2,4-bis(dodecyloxy)cyclohexylmethylamino group, a 3,5-bis[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group, or a 3,4,5-tris[3',4',5'-tris(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino group. is a base, or LY is a single bond or succinyl-1,4-piperazinediyl group, and Z is a 3,4,5-tris(octadecyloxy)benzoyl group. It is the basis.

[0153] The protecting group (Pg-5) is particularly preferred, L is a succinyl group, and YZ is a 3,4,5-tris(octadecyloxy)benzyloxy group, a 3,4,5-tris(octadecyloxy)cyclohexylmethyloxy group, or a phenyl(2,3,4-tris(octadecyloxy)phenyl)methylamino group. is a base, or LY is a succinyl-1,4-piperazinediyl group, and Z is a 3,4,5-tris(octadecyloxy)benzoyl group. It is the basis.

[0154] The protecting group (Pg-5) is most preferably, L is a succinyl group, and YZ is either a 3,4,5-tris(octadecyloxy)benzyloxy group or a 3,4,5-tris(octadecyloxy)cyclohexylmethyloxy group. is a base, or LY is a succinyl-1,4-piperazinediyl group, and Z is a 3,4,5-tris(octadecyloxy)benzoyl group. It is the basis.

[0155] Protecting groups (Pg-5) and compounds used to form them can be formed or produced by known methods (for example, the methods described in International Publication No. 2017 / 104836, International Publication No. 2019 / 131719, International Publication No. 2020 / 235658, International Publication No. 2021 / 039935 or International Publication No. 2021 / 198883) or similar methods.

[0156] 1-2. Terminology related to oligonucleotide synthesis In this specification, the term "solid support" is not particularly limited as long as it is used in the art for solid-phase synthesis of oligonucleotides, and includes, for example, glass beads, resin beads, etc. The support or resin used as the solid support can be any support or resin that is known in the art and suitable for use in solid-phase synthesis. Examples of such supports or resins include polystyrene supports (polystyrene supports may be further functionalized with, for example, p-methylbenzylhydrylamine), diatomaceous earth-encapsulated polydimethylacrylamide (pepsin K), silica, microporous glass, amphiphilic polystyrene-polyethylene glycol (PEG) resin, PEG-polyamide, PEG-polyester resin, Wang-PEG resin, Rink-amide PEG resin, etc. The support or resin may be modified. Examples of modified supports or resins include long-chain alkylamino controlled pore glass (LCAA CPG).

[0157] In this specification, "nucleoside," which is a constituent unit of oligonucleotides, means a compound in which a nucleic acid base is bonded to the 1-position of a sugar (for example, 2-deoxyribose, ribose, 2-deoxyribose or ribose in which the 2- and 4-carbon atoms are linked by a divalent organic group, 2-deoxyribose or ribose in which the 3- and 5-carbon atoms are linked by a divalent organic group, 2-deoxyribose or ribose in which the 3- and 4-carbon atoms are linked by a divalent organic group, etc.) by N-glycosidation.

[0158] In this specification, "nucleic acid base" is not particularly limited as long as it is used in the synthesis of nucleic acids, and examples include pyrimidine bases such as cytosyl groups (cytosine bases), uracil groups (uracil bases), and thyminyl groups (thymine bases), and purine bases such as adenyl groups (adenine bases) and guanyl groups (guanine bases). Furthermore, "nucleic acid base that may be protected" means, for example, that the amino group, carbonyl group, etc. in the nucleic acid base may be protected. A nucleic acid base that may be protected is preferably a nucleic acid base in which the amino group may be protected by the aforementioned amino group protecting group, more preferably a nucleic acid base that does not have an amino group, or a nucleic acid base that has an amino group protected by the aforementioned amino group protecting group.

[0159] The carbonyl group can be protected by reacting it with, for example, phenol, 2,5-dichlorophenol, 3-chlorophenol, 3,5-dichlorophenol, 2-formylphenol, 2-naphthol, 4-methoxyphenol, 4-chlorophenol, 2-nitrophenol, 4-nitrophenol, 4-acetylaminophenol, pentafluorophenol, 4-pivaloyloxybenzyl alcohol, 4-nitrophenethyl alcohol, 2-(methylsulfonyl)ethanol, 2-(phenylsulfonyl)ethanol, 2-cyanoethanol, 2-(trimethylsilyl)ethanol, dimethylcarbamate chloride, diethylcarbamate chloride, ethylphenylcarbamate chloride, 1-pyrrolidinecarboxylic acid chloride, 4-morpholinecarboxylic acid chloride, diphenylcarbamate chloride, etc.

[0160] As used herein, "nucleic acid base" also includes modified nucleic acid bases in which the nucleic acid base is substituted with substituents (e.g., halogen atoms, alkyl groups, aralkyl groups, alkoxy groups, acyl groups, alkoxyalkyl groups, hydroxyl groups, amino groups, monoalkylamino groups, dialkylamino groups, carboxyl groups, cyano groups, nitro groups, etc.) (e.g., 8-bromoadenyl group, 8-bromoguanyl group, 5-bromocytosyl group, 5-iodocytosyl group, 5-bromouracil group, 5-iodouracil group, 5-fluorouracil group, 5-methylcytosyl group, 8-oxoguanyl group, hypoxanthinyl group, etc.).

[0161] In this specification, "sugar" includes amino sugars in which a hydroxyl group is replaced by an amino group, and ribose in which the hydroxyl group at position 2 is replaced by a halogen atom.

[0162] Examples of amino sugars include 2-deoxyribose, in which the hydroxyl group at position 3 is replaced by an amino group, ribose, in which the hydroxyl group at position 3 is replaced by an amino group, and ribose, in which the hydroxyl group at position 3 is replaced by an amino group and the hydroxyl group at position 2 is replaced by a halogen (in the formula below, X s (This indicates a halogen atom.)

[0163] [ka]

[0164] Examples of 2-deoxyribose or ribose in which the 2- and 4-carbon atoms are bonded by a divalent organic group, 2-deoxyribose or ribose in which the 3- and 5-carbon atoms are bonded by a divalent organic group, or 2-deoxyribose or ribose in which the 3- and 4-carbon atoms are bonded by a divalent organic group include the following compounds.

[0165] [ka]

[0166] [In the formula, R represents a hydrogen atom, an optionally substituted hydrocarbon group, an optionally substituted hydroxyl group, or an optionally substituted amino group, and R' represents a hydrogen atom or a hydroxyl group.]

[0167] In this specification, "phosphate group" includes not only -OP(=O)(OH)2 but also groups in which the oxygen atom is replaced by a sulfur atom or NH (e.g., -OP(=S)(OH)2, -NH-P(=O)(OH)2, -NH-P(=S)(OH)2). Furthermore, the hydroxyl group (-OH) in the phosphate group may be replaced by -OR. p (In the formula, R p The term "phosphate group" also includes groups that have been replaced by organic groups (e.g., protecting groups for phosphate groups) (e.g., protected phosphate groups).

[0168] In this specification, "phosphorous ester bond" refers to a bond represented by the following formula (P1-1), or a bond represented by the following formula (P1-2) in which the oxygen atom in the bond is replaced by NH (where * indicates the bond position).

[0169] [ka]

[0170] In this specification, "phosphonic acid ester bond" refers to a bond represented by the following formula (P2-1), or a bond represented by the following formula (P2-2) in which the oxygen atom in the bond is replaced by NH (where * indicates the bond position).

[0171] [ka]

[0172] In this specification, "phosphate ester bond" refers to a bond represented by the following formula (P3-1), or a bond represented by the following formula (P3-2) in which the oxygen atom in the said bond is replaced by NH (where * indicates the bond position).

[0173] [ka]

[0174] In this specification, "thiophosphate ester bond" refers to a bond represented by the following formula (P4-1), or a bond represented by the following formula (P4-2) in which the oxygen atom in the bond is replaced by NH (where * indicates the bond position).

[0175] [ka]

[0176] In this specification, "nucleotide" refers to a compound in which a phosphate group is bonded to a nucleoside. The explanation of the phosphate group is as described above.

[0177] In this specification, "oligonucleotide" means a compound in which one or more nucleotides are linked to a nucleoside. The term "oligonucleotide" includes not only oligonucleotides having a phosphate ester bond represented by formula (P3-1), but also oligonucleotides having a phosphate ester bond represented by formula (P3-2), oligonucleotides having a thiophosphate ester bond represented by formula (P4-1), and oligonucleotides having a thiophosphate ester bond represented by formula (P4-2). The number of nucleosides in the oligonucleotide in this invention is not particularly limited, but is preferably 2 to 50, more preferably 2 to 30.

[0178] In this specification, "phosphoramidite" means a monoamide of a phosphite diester (P(OR)2(NR2), where each of the four Rs independently represents an optionally substituted alkyl group, and the NR2 may have two Rs bonded to each other to form a cyclic amino group).

[0179] In this specification, "phosphoramiditized nucleoside" means a nucleoside with -X n -P(OR)(NR2) is a base (where X nThe compound is obtained by introducing (where R represents an oxygen atom or NH, each of the three Rs independently represents an optionally substituted alkyl group, and the NR2 may be formed by two Rs bonding to each other to form a cyclic amino group).

[0180] In this specification, “phosphoramiditized nucleotide” means a nucleotide with -X n -P(OR)(NR2) is a base (where X n (where is an oxygen atom or NH, the three Rs each independently represent an optionally substituted alkyl group, and the NR2 may be obtained by introducing two Rs bonded to each other to form a cyclic amino group) -X n This refers to compounds having a group represented by -P(OR)(NR2) and a phosphate group. The phosphate group may be substituted.

[0181] In this specification, "phosphoramiditized oligonucleotide" means an oligonucleotide with -X n -P(OR)(NR2) is a base (where X n The compound is obtained by introducing (where R represents an oxygen atom or NH, each of the three Rs independently represents an optionally substituted alkyl group, and the NR2 may be formed by two Rs bonding to each other to form a cyclic amino group).

[0182] In this specification, "oligonucleotide precursor having phosphite ester bonds" means a precursor in which the phosphate ester bonds in the oligonucleotide are replaced by phosphite ester bonds. By oxidizing this precursor, oligonucleotides having phosphate ester bonds can be produced.

[0183] Oligonucleotide precursors having phosphite ester bonds are preferably (1) Oligonucleotide precursors having phosphite bonds, obtained by condensation of a nucleoside, nucleotide, or oligonucleotide with a phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide, or (2) Oligonucleotide precursors having phosphite bonds obtained by condensation of oligonucleotides with phosphoramidites That is the case.

[0184] Oligonucleotide precursors having phosphite ester bonds are more preferably oligonucleotide precursors having phosphite ester bonds obtained by condensation of a nucleoside, nucleotide, or oligonucleotide with a phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide.

[0185] The oligonucleotide precursor having a phosphite ester bond is more preferably obtained by condensation of a nucleoside, nucleotide, or oligonucleotide (a) having a hydrophobic protecting group with a nucleoside, nucleotide, or oligonucleotide (b) whose hydroxyl group is protected by a temporary protecting group that can be removed under acidic conditions and is phosphoramidized.

[0186] The aforementioned condensation may be either a condensation in which the oligonucleotide chain is extended from the 3' end to the 5' end (hereinafter referred to as "3'-5' condensation") or a condensation in which the oligonucleotide chain is extended from the 5' end to the 3' end (hereinafter referred to as "5'-3' condensation"). The following describes, in order, the preferred nucleoside, nucleotide, or oligonucleotide (a) and the phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide (b) for each of the 3'-5' condensation and 5'-3' condensation.

[0187] In the 3'-5' condensation, the nucleoside, nucleotide, or oligonucleotide (a) may be, for example, formula (aI):

[0188] [ka]

[0189] [In the formula, r represents a non-negative integer; r+1 Base 1 Each of these independently represents a nucleic acid base that may be protected; r+1 X n1 Each of these independently represents either an oxygen atom or NH; r+1 X n2 Each of these independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group bonded to the 2- and 4-carbon atoms; r R 10 Each of these independently represents either an oxygen atom or a sulfur atom; r R p1 Each of these independently represents a protecting group for the phosphate group; Pg is **LYZ(where ** is X) n1 (Indicates the bond position with), **LY-Sp(where ** is X) n1 The bond position is indicated by, and Sp indicates a solid support) or indicates a solid support; and The definitions and explanations of L, Y, and Z are the same as those for the protecting group (Pg-5) described above. Examples include compounds represented by (i.e., nucleosides or oligonucleotides).

[0190] When r is 0, compound (aI) is a nucleoside, and when r is 1 or greater, compound (aI) is an oligonucleotide. r is preferably 49 or less, more preferably 29 or less, even more preferably 19 or less, particularly preferably 4 or less, and most preferably 2 or less.

[0191] Nucleic acid bases 1The amino group is preferably protected by a protecting group. Examples of amino group protecting groups include the amino group protecting groups mentioned above, but Base 1 The protecting group for the amino group is preferably an acetyl group, a phenoxyacetyl group, a 4-isopropylphenoxyacetyl group, a benzoyl group, an isobutyryl group, a (2-hexyl)decanoyl group, a dimethylformamidinyl group, and =NC(R 11 )-N(R 12 )(R 13 ) is expressed as (where R 11 R indicates a methyl group. 12 and R 13 Each of them is independently C 1-6 Indicates an alkyl group, or R 11 and R 12 These may combine to form a 5- or 6-membered nitrogen-containing heterocycle with the carbon and nitrogen atoms to which they are bonded. If compound (aI) has multiple amino groups, there may be only one protecting group for the amino group, or there may be two or more.

[0192] X n2 The halogen atom is preferably a fluorine atom or a chlorine atom, with a fluorine atom being more preferred.

[0193] X n2 Examples of hydroxyl group protecting groups that may be protected include the aforementioned hydroxyl group protecting groups, but are preferably methyl group, 2-methoxyethyl group, triethylsilyl group, triisopropylsilyl group, or tert-butyldimethylsilyl group, (2-cyanoethoxy)methyl group, or 1-(2-cyanoethoxy)ethyl group.

[0194] X n2 The "divalent organic group bonded to the 2- and 4-carbon atoms" is not particularly limited as long as it is bonded to the 2- and 4-carbon atoms of the nucleoside. Examples of this divalent organic group include a substituted C 2-7 Alkylene groups, as well as -O- and -NR groups. 33 -(R 33 is a hydrogen atom or C 1-6(Indicating an alkyl group), -S-, -CO-, -COO-, -OCONR 34 -(R 34 is a hydrogen atom or C 1-6 (Indicating an alkyl group) and -CONR 35 -(R 35 is a hydrogen atom or C 1-6 A divalent linker selected from alkyl groups, and a C which may be substituted. 1-7 Examples include divalent organic groups composed of alkylene groups. 1-7 Alkylene group and C 2-7 Examples of substituents that the alkylene group may have include a methylidene group (CH2=).

[0195] The "divalent organic group bonded to the 2- and 4-carbon atoms" may be a substituted C. 2-7 Alkylene group, -OR i -(R i C is bonded to the 4th carbon atom. 1-6 (showing an alkylene group), -O-NR 33 -R j -(R j C is bonded to the 4th carbon atom. 1-6 It shows an alkylene group, R 33 (This indicates the same meaning as above), -OR k -OR l -(R k is C 1-6 It shows an alkylene group, R l C is a carbon that forms a bridge bond with the carbon atom at position 4. 1-6 (showing an alkylene group) is preferred, -OR i -(R i (This indicates the same meaning as above), -O-NR 33 -R j -(R j and R 33 (This indicates the same meaning as above), -OR k -OR l -(R k and R l (This indicates the same meaning as above) is more preferable. i , R j , R k and R l C shown by1-6 The alkylene group is preferably either a methylene group or an ethylene group, independently of each other.

[0196] Examples of "divalent organic groups bonded to the 2- and 4-carbon atoms" include -O-CH2-, -O-CH2-CH2-, and -O-NR 33 -CH2-(R 33 -O-CH2-O-CH2- is more preferred (where -O-CH2-, -O-CH2-CH2-, -O-NH-CH2-, -ON(CH3)-CH2-, and -O-CH2-O-CH2- (where the left side is bonded to the 2-carbon atom and the right side is bonded to the 4-carbon atom).

[0197] r+1 X n2 Each of these is independently preferably a hydrogen atom, a halogen atom, or an optionally protected hydroxyl group, and more preferably a hydrogen atom, a fluorine atom, or an optionally protected hydroxyl group. r R p1 Examples include the aforementioned phosphate group protecting group, but a 2-cyanoethyl group is preferred.

[0198] In the above equation (aI), Pg is **LY-Sp (where ** is X). n1 The formula indicates the binding position with (wherein Sp represents the solid support) or, if it is a solid support, the compound (aI) is a nucleoside or oligonucleotide supported on the solid support. Pg is preferably **LYZ (wherein ** is X n1 (Indicating the bonding position with) or a solid support, more preferably **LYZ.** The definitions and descriptions of L, Y, and Z are the same as those for the protecting group (Pg-5) described above.

[0199] In the 3'-5' condensation, the phosphoramidized nucleoside, nucleotide, or oligonucleotide (b) is, for example, Equation (bI):

[0200] [ka]

[0201] [In the formula, s represents a non-negative integer; s+1 Bases 2 Each of these independently represents a nucleic acid base that may be protected; s+1 X n3 Each of these independently represents either an oxygen atom or NH; s+1 X n4 Each of these independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group bonded to the 2- and 4-carbon atoms; "Q" indicates a temporary protecting group that can be removed under acidic conditions of the hydroxyl group; s R 14 Each of these independently represents either an oxygen atom or a sulfur atom; s+1 R p2 Each of these independently represents a protecting group for the phosphate group; and R 15 and R 16 Each of these independently represents either an alkyl group or a 5- or 6-membered saturated cyclic amino group formed together with an adjacent nitrogen atom, and such saturated cyclic amino group may have one oxygen atom or sulfur atom as a ring constituent atom in addition to the nitrogen atom. Examples include compounds represented by (i.e., nucleosides or oligonucleotides).

[0202] When s is 0, compound (bI) is a phosphoramidite-treated nucleoside, and when s is 1 or greater, compound (bI) is a phosphoramidite-treated oligonucleotide. In compound (bI), s is preferably 49 or less, more preferably 29 or less, even more preferably 19 or less, particularly preferably 4 or less, and most preferably 2 or less.

[0203] Nucleic acid bases 2The amino group is preferably protected by a protecting group. In addition to the aforementioned amino group protecting groups, the aforementioned -LYZ group can be used as the protecting group. The protecting group is preferably the aforementioned amino group protecting group, and more preferably an acetyl group, phenoxyacetyl group, 4-isopropylphenoxyacetyl group, benzoyl group, isobutyryl group, (2-hexyl)decanoyl group, dimethylformamidinyl group, and =NC(R 11 )-N(R 12 )(R 13 A base represented by (wherein R in the formula) 11 R indicates a methyl group. 12 and R 13 Each of them is independently C 1-6 Indicates an alkyl group, or R 11 and R 12 These groups may combine to form a 5- or 6-membered nitrogen-containing heterocycle with the carbon and nitrogen atoms to which they are bonded. If compound (bI) has multiple amino groups, there may be only one protecting group for the amino group, or there may be two or more.

[0204] X in equation (bI) above n4 The explanation is that X in equation (aI) above n2 This is the same as the explanation. s+1 X n4 Each of these is independently preferably a hydrogen atom, a halogen atom, or an optionally protected hydroxyl group, and more preferably a hydrogen atom or an optionally protected hydroxyl group.

[0205] R in equation (bI) above p2 The explanation is that R in equation (aI) above p1 This is the same as the explanation. s+1 R p2 Examples include the aforementioned phosphate group protecting group, but a 2-cyanoethyl group is preferred.

[0206] R in equation (bI) above 15 and R 16 Each of these is independently, preferably C 1-10It is an alkyl group or a 5 or 6-membered saturated cyclic amino group formed together with an adjacent nitrogen atom, more preferably C 1-10 It is an alkyl group, and more preferably C 1-6 It is an alkyl group.

[0207] Compound (aI) and compound (bI) can be produced by known methods (e.g., the method described in International Publication No. 2017 / 104836) or similar methods.

[0208] The oligonucleotide precursor having a phosphite ester bond, obtained by the condensation of compound (aI) and compound (bI), is the compound represented by the following formula (cI) (the definition and explanation of the symbols in formula (cI) below are as described above).

[0209] [ka]

[0210] In the above equation (cI), Pg is **LY-Sp (where ** is X). n1 The formula indicates the binding position with (wherein Sp represents the solid support) or, if it is a solid support, compound (cI) is an oligonucleotide precursor supported on the solid support. Pg is preferably **LYZ (wherein ** is X n1 (Indicating the bonding position with) or a solid-phase support, more preferably LYZ.

[0211] In the above equation (cI), s R 14 and r R 10 Preferably, at least one of them is a sulfur atom.

[0212] In the 5'-3' condensation, the nucleoside, nucleotide, or oligonucleotide (a) is, for example, formula (a-I'):

[0213] [ka]

[0214] [In the formula, r represents a non-negative integer; r+1 Base 1 Each of these independently represents a nucleic acid base that may be protected; r X n1 Each of these independently represents either an oxygen atom or NH; r+1 X n2 Each of these independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group bonded to the 2- and 4-carbon atoms; X n5 This indicates a hydroxyl group or an amino group; r R 10 Each of these independently represents either an oxygen atom or a sulfur atom; r R p1 Each of these independently represents a protecting group for the phosphate group; Pg is **LYZ (wherein ** indicates the bond position with the oxygen atom), **LY-Sp (wherein ** is X) n1 The bond position is indicated by, and Sp indicates a solid support) or indicates a solid support; and The definitions and explanations of L, Y, and Z are the same as those for the protecting group (Pg-5) described above. Examples include compounds represented by (i.e., nucleosides or oligonucleotides).

[0215] X in equation (a-I') n5 It is preferably a hydroxyl group. n5 The explanations for the other symbols are the same as those for the symbols in formula (aI) above.

[0216] In the above equation (a-I'), Pg is **LY-Sp (where ** is X). n1If the compound (a-I') is a solid support (wherein Sp indicates the binding position with the oxygen atom), then the compound (a-I') is a nucleoside or oligonucleotide supported on the solid support. Pg is preferably a LYZ (wherein ** indicates the binding position with the oxygen atom) or a solid support, and more preferably a LYZ.

[0217] In the 5'-3' condensation, the phosphoramidized nucleoside, nucleotide, or oligonucleotide (b) is, for example, formula (b-I'):

[0218] [ka]

[0219] [In the formula, s represents a non-negative integer; s+1 Bases 2 Each of these independently represents a nucleic acid base that may be protected; s pieces of X n3 Each of these independently represents either an oxygen atom or NH; s+1 X n4 Each of these independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group bonded to the 2- and 4-carbon atoms; X n6 This indicates a hydroxyl group protected by a temporary protecting group that can be removed under acidic conditions, or an amino group protected by a temporary protecting group that can be removed under acidic conditions; s R 14 Each of these independently represents either an oxygen atom or a sulfur atom; s+1 R p2 Each of these independently represents a protecting group for the phosphate group; and R 15 and R 16 Each of these independently represents either an alkyl group or a 5- or 6-membered saturated cyclic amino group formed together with an adjacent nitrogen atom, and such saturated cyclic amino group may have one oxygen atom or sulfur atom as a ring constituent atom in addition to the nitrogen atom. Examples of compounds represented by [the formula shown] are given.

[0220] X n6 This is a hydroxyl group that is preferably protected by a temporary protecting group that can be removed under acidic conditions. n6 The explanations for the other symbols are the same as those for the symbols in formula (bI) above.

[0221] Compounds (a-I') and (b-I') can be produced by known methods (e.g., the method described in International Publication No. 2017 / 104836) or similar methods.

[0222] The oligonucleotide precursor having a phosphite ester bond, obtained by the condensation of compound (a-I') and compound (b-I'), is given by the following formula (c-I'):

[0223] [ka]

[0224] [In the formula, r represents a non-negative integer; r+1 Base 1 Each of these independently represents a nucleic acid base that may be protected; r+1 X n1 Each of these independently represents either an oxygen atom or NH; r+1 X n2 Each of these independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group bonded to the 2- and 4-carbon atoms; r R 10 Each of these independently represents either an oxygen atom or a sulfur atom; r R p1 Each of these independently represents a protecting group for the phosphate group; s represents a non-negative integer; s+1 Bases 2 Each of these independently represents a nucleic acid base that may be protected; s pieces of X n3Each of these independently represents either an oxygen atom or NH; s+1 X n4 Each of these independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group bonded to the 2- and 4-carbon atoms; X n6 This indicates a hydroxyl group protected by a temporary protecting group that can be removed under acidic conditions, or an amino group protected by a temporary protecting group that can be removed under acidic conditions; s R 14 Each of these independently represents either an oxygen atom or a sulfur atom; s+1 R p2 Each of these independently represents a protecting group for the phosphate group; Pg is **LYZ (wherein ** indicates the bond position with the oxygen atom), **LY-Sp (wherein ** is X) n1 The bond position is indicated by, and Sp indicates a solid support) or indicates a solid support; and The definitions and explanations of L, Y, and Z are the same as those for the protecting group (Pg-5) described above. It is a compound represented by (the symbols in the formula are explained above).

[0225] In the above equation (c-I'), Pg is **LY-Sp (where ** is X). n1 If the compound (c-I') is a solid support (wherein Sp indicates the binding position with the oxygen atom), then the compound (c-I') is an oligonucleotide precursor supported on the solid support. Pg is preferably a LYZ (wherein ** indicates the binding position with the oxygen atom) or a solid support, and more preferably a LYZ.

[0226] In the above equation (c-I'), r R 10 and s R 14 Preferably, at least one of them is a sulfur atom.

[0227] Next, we will describe an oligonucleotide precursor having a phosphite ester bond obtained by the condensation of an oligonucleotide with a phosphoramidite. Examples of such precursors include formula (γ-I) or formula (γ-I'):

[0228] [ka]

[0229] [In the formula, t represents an integer greater than or equal to 1. t+1 Bases 1 Each of these independently represents a nucleic acid base that may be protected; t+1 X n1 Each of these independently represents either an oxygen atom or NH; t+1 X n2 Each of these independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group bonded to the 2- and 4-carbon atoms; t R 10 Each of these independently represents either an oxygen atom or a sulfur atom; t R p1 and R p3 Each of these independently represents a protecting group for the phosphate group; R p4 This indicates an alkyl group having a protected amino group, or a protecting group for a phosphate group; Pg is **LYZ(in the formula, ** is X n1 (Indicates the bonding position with or the bonding position with the oxygen atom), **LY-Sp(wherein ** is X n1 The bond position is indicated by, and Sp indicates a solid support) or indicates a solid support; and The definitions and explanations of L, Y, and Z are the same as those for the protecting group (Pg-5) described above. Examples of compounds represented by [the formula shown] are given.

[0230] In the above formula (γ-I) or formula (γ-I'), Pg is **LY-Sp (where ** is X). n1If the compound (γ-I) or compound (γ-I') is a solid support (wherein Sp indicates the binding position with the oxygen atom), then Pg is preferably a LYZ (wherein ** indicates the binding position with the oxygen atom) or a solid support, and more preferably a LYZ.

[0231] The symbols in formulas (γ-I) and (γ-I') above will be explained. t is preferably 49 or less, more preferably 29 or less, even more preferably 19 or less, particularly preferably 4 or less, and most preferably 2 or less. p3 and R p4 Examples of "phosphate protecting groups" include the aforementioned phosphate protecting groups. p3 The group is preferably a 2-cyanoethyl group. p4 Examples of the "protected amino group" in the "alkyl group having a protected amino group" include an amino group protected by the aforementioned amino group protecting group. p4 The "alkyl group having a protected amino group" is preferably a C group having a protected amino group. 1-6 It is an alkyl group, more preferably a 6-(trifluoroacetylamino)hexyl group. p4 This is preferably a 2-cyanoethyl group or a 6-(trifluoroacetylamino)hexyl group. The explanations for the other symbols are as described above.

[0232] In this specification, "phosphonate ester bond-containing oligonucleotide precursor" means a precursor in which the phosphate ester bond in the oligonucleotide is replaced by a phosphonate ester bond. By oxidizing this precursor, an oligonucleotide containing a phosphate ester bond can be produced.

[0233] The oligonucleotide precursor having a phosphonic acid ester bond is preferably obtained by condensation of a nucleoside, nucleotide, or oligonucleotide (a) having a hydrophobic protecting group with an H-phosphonate (B) protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions. The H-phosphonate (B) may be used in the form of a salt such as a triethylamine salt.

[0234] The condensation may be either a 3'-5' condensation or a 5'-3' condensation. Preferred nucleosides, nucleotides, or oligonucleotides (a) and H-phosphonates (B) for each of the 3'-5' and 5'-3' condensations will be described in order.

[0235] In the 3'-5' condensation, the nucleoside, nucleotide, or oligonucleotide (a) is preferably the aforementioned compound (aI).

[0236] In the 3'-5' condensation, the H-phosphonate (B) can be, for example, a compound represented by the following formula (BI) (the definitions and explanations of the symbols in formula (BI) below are the same as those for the symbols in formula (bI) above).

[0237] [ka]

[0238] Compound (BI) can be produced by known methods (e.g., the method described in Acta Biochimica Polonica. 1998, 45, 907-915.) or similar methods.

[0239] The oligonucleotide precursor having a phosphonic acid ester bond obtained by the condensation of compound (aI) and compound (BI) is a compound represented by the following formula (CI) (the definitions and explanations of the symbols in formula (CI) below are the same as those for the symbols in formula (cI) above).

[0240] [ka]

[0241] In the above formula (CI), Pg is **LY-Sp (where ** is X). n1 The formula indicates the binding position with (wherein Sp represents the solid support) or, if it is a solid support, the compound (CI) is an oligonucleotide precursor supported on the solid support. Pg is preferably **LYZ (wherein ** is X n1 (Indicating the bonding position with) or a solid-phase support, more preferably LYZ.

[0242] In the above formula (CI), s R 14 and r R 10 Preferably, at least one of them is a sulfur atom.

[0243] In the 5'-3' condensation, the nucleoside, nucleotide, or oligonucleotide (a) is preferably the aforementioned compound (a-I').

[0244] In the 5'-3' condensation, examples of H-phosphonate (B) include compounds represented by the following formula (B-I') (the definitions and explanations of the symbols in formula (B-I') below are the same as those for the symbols in formula (b-I') above).

[0245] [ka]

[0246] Compound (B-I') can be prepared by known methods (for example, the method described in Phosphorus Chemistry II. Topics in Current Chemistry, vol 361. Springer, Cham. Recent Advances in H-Phosphonate Chemistry) or similar methods.

[0247] The oligonucleotide precursor having a phosphonic acid ester bond, obtained by the condensation of compound (a-I') and compound (B-I'), is the compound represented by the following formula (C-I') (the definitions and explanations of the symbols in formula (C-I') below are the same as those for the symbols in formula (c-I') above).

[0248] [ka]

[0249] In the above equation (C-I'), Pg is **LY-Sp (where ** is X). n1 If the compound (CI) is a solid support (wherein Sp indicates the binding position with the oxygen atom) or a solid support, then the compound (CI) is an oligonucleotide precursor supported on the solid support. Pg is preferably a LYZ (wherein ** indicates the binding position with the oxygen atom) or a solid support, and more preferably a LYZ.

[0250] In the above formula (C-I'), r R 10 and s R 14 Preferably, at least one of them is a sulfur atom.

[0251] In this specification, "one-pot synthesis" means a multi-step synthesis comprising steps for synthesizing an intermediate and a step for synthesizing the desired final product, wherein the intermediate is not isolated. The "step for synthesizing an intermediate" may be one or more.

[0252] 2. Manufacturing method of the present invention The manufacturing method of the present invention will be described below. The manufacturing method of the present invention uses the following formula (I) as an oxidizing agent:

[0253] [ka]

[0254] The present invention is characterized by using a compound represented by (excluding 2,2-dipyridyl disulfide) to oxidize an oligonucleotide precursor having a phosphite ester bond or a phosphonic acid ester bond to produce an oligonucleotide having a phosphate ester bond. Compound (I) may be used alone or in combination of two or more.

[0255] 2-1. Oxidizing agents The following describes compound (I) (excluding 2,2-dipyridyl disulfide), which is an oxidizing agent used in the manufacturing method of the present invention.

[0256] X in equation (I) above 1 This is a single bond, sulfur atom, oxygen atom, -S(=O)2- or -N(-R 3 )-is.

[0257] X 1 If it is a single bond, then R in formula (I) 1 is a halogen atom, and R in formula (I) 2 This is an optionally substituted aryl group or an optionally substituted heteroaryl group. X 1 If is a sulfur atom or -S(=O)2-, then R in formula (I) 1 R in formula (I) is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 This is an optionally substituted aryl group or an optionally substituted heteroaryl group. X 1 If is an oxygen atom, then R in formula (I) 1 and R 2 These, along with the sulfur and oxygen atoms to which they are bonded, form a heterocycle that may be substituted. X 1 -N(-R 3 )- If that is the case, (1) R in formula (I) above 1R in formula (I) is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 and R 3 They, together with the sulfur and nitrogen atoms to which they bond, form a heterocycle which may be substituted, or (2) R in formula (I) above 1 and R 3 Together with the nitrogen atoms to which they are bonded, they form a heterocycle which may be substituted, and R in formula (I) 2 This is an optionally substituted aryl group or an optionally substituted heteroaryl group.

[0258] 2-1-1. Compound (IA) and Compound (IB) Among compound (I), In equation (I) above, X 1 -N(-R 3 )- and R 1 However, it is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 and R 3 These, together with the sulfur and nitrogen atoms to which they are bonded, form compounds (referred to herein as "compound (IA)") which may be substituted, and In equation (I) above, X 1 However, it is a sulfur atom, R 1 However, it is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 However, compounds that have an optionally substituted aryl group or an optionally substituted heteroaryl group (excluding 2,2-dipyridyl disulfide) (hereinafter referred to as "compound (IB)") It is preferable.

[0259] In other words, among compound (I), X 1 However, sulfur atoms or -N(-R 3 )- and, R 1However, it is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. X 1 If R is a sulfur atom, 2 is an optionally substituted aryl group or an optionally substituted heteroaryl group, and X 1 -N(-R 3 )- If R 2 and R 3 These, along with the sulfur and nitrogen atoms to which they bond, form a heterocycle that may be substituted. Compounds (excluding 2,2-dipyridyl disulfide) are preferred. Below, we will first explain compound (IA) and compound (IB).

[0260] 2-1-1A. Compound (IA) First, let's explain compound (IA). In compound (IA), R in formula (I) 1 is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, preferably an optionally substituted C 1-10 The group is an alkyl group, an optionally substituted phenyl group, an optionally substituted 5- or 6-membered monocyclic heteroaryl group, or an optionally substituted 8- to 14-membered fused polycyclic heteroaryl group.

[0261] In compound (IA), R in formula (I) 2 and R 3 These, along with the sulfur and nitrogen atoms to which they are bonded, form a heterocycle that may be substituted.

[0262] 2-1-1A-1. Compound (Ia) Compound (IA) is preferably a compound represented by the following formula (Ia). Compound (Ia) may be used alone or in combination of two or more types.

[0263] [ka]

[0264] The following describes each group of compound (Ia). R in equation (Ia) above 1a is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, preferably an optionally substituted C 1-10 The group is an alkyl group, an optionally substituted phenyl group, or an optionally substituted 5- or 6-membered monocyclic heteroaryl group.

[0265] In equation (Ia), ring A is a 5- or 6-membered unsaturated hetero ring. 1a is -S(=O)-, -S(=O)2-, -C(=O)-, or -C(=S)-, preferably -C(=O)-.

[0266] In equation (Ia) above, m is an integer from 0 to 3, and m R 2a Each of these is independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, preferably even if substituted. 1-6 A good alkyl group, may be substituted C 1-6 It is an alkoxy group or an electron-withdrawing group. Here, "m is 0" means R 2a This means that it does not exist.

[0267] If m in equation (Ia) is an integer greater than or equal to 2, then two adjacent R 2a Together with ring A, it may form a fused ring that may be substituted.

[0268] A 5- or 6-member unsaturated hetero ring A is a ring represented by one of the following equations (r2) to (r4) [the definitions of the symbols in the equations are as described above]. Note that in equations (r2) to (r4) below, R 2a It has been omitted.

[0269] [ka]

[0270] Two adjacent R 2a The fused ring formed by ring A is preferably a bicyclic or tricyclic fused ring, and more preferably a bicyclic fused ring. Examples of the fused ring include rings represented by any of the following formulas (r5) to (r25). The fused ring may be substituted.

[0271] [ka]

[0272] [ka]

[0273] 2-1-1A-2. Compound (Ic) Compound (Ia) is preferably a compound represented by the following formula (Ic).

[0274] [ka]

[0275] The following describes each group of compound (Ic). R in equation (Ic) 1c is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, preferably an optionally substituted C 1-6 The group is an alkyl group, an optionally substituted phenyl group, or an optionally substituted 5- or 6-membered monocyclic heteroaryl group.

[0276] In one embodiment of the manufacturing method of the present invention, R 1c Preferably (1) A C group which may be substituted with one or two substituents selected from the group consisting of a substituted phenyl group and a substituted 5 or 6-membered monocyclic heteroaryl group.1-6 alkyl group, (2) A phenyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (3) A 5 or 6-membered monocyclic heteroaryl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. That is the case.

[0277] In another embodiment of the manufacturing method of the present invention, R 1c Preferably (1) A C group which may be substituted with one or two selected substituents selected from the group consisting of optionally substituted phenyl groups and optionally substituted 5 or 6-membered monocyclic heteroaryl groups. 1-6 alkyl group, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A phenyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. That is the case.

[0278] In another embodiment of the manufacturing method of the present invention, R 1c Preferably (1) C which may be substituted with one or two pyridyl groups 1-6 alkyl group, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A phenyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups, or (3) C may not be substituted 1-6Alkyl alkyl groups, or substituted C 1-6 A pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups. That is the case.

[0279] In another embodiment of the manufacturing method of the present invention, R 1c Preferably (1) C which may be substituted with one or two pyridyl groups 1-6 alkyl group, (2) One or two C 1-6 A phenyl group which may be substituted with an alkyl group, or (3) Pyridyl group (especially 2-pyridyl group or 4-pyridyl group) That is the case.

[0280] In another embodiment of the manufacturing method of the present invention, R 1c Preferably (1)C 1-6 alkyl group, (2)C 1-6 Alkyl alkyl group, C 1-6 A phenyl group which may be substituted with one or two substituents selected from the group consisting of alkoxy groups and halogen atoms, (3) One C 1-6 Pyridyl groups substituted with alkyl groups (e.g., 4-methyl-2-pyridyl group) That is the case.

[0281] Y in equation (Ic) above 1c is -S(=O)-, -S(=O)2-, -C(=O)-, or -C(=S)-, preferably -C(=O)-.

[0282] In the above formula (Ic), p is 0 or 1, preferably 0. Here, "p is 0" means that -CH(R 2c )- means that it does not exist.

[0283] R in equation (Ic) 2cis a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, preferably a hydrogen atom or an optionally substituted C 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group or an electron-withdrawing group.

[0284] In formula (Ic), ring C is a 6-membered aromatic hydrocarbon ring, a 6-membered nitrogen-containing aromatic heterocycle, or a 10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom. In one embodiment of the manufacturing method of the present invention, the ring C is preferably a 6-membered aromatic hydrocarbon ring (i.e., a benzene ring) or a 6-membered nitrogen-containing aromatic heterocycle, more preferably a benzene ring or a pyridine ring, and even more preferably a pyridine ring. In another embodiment of the manufacturing method of the present invention, the ring C is preferably a benzene ring, a pyridine ring, or a quinoline ring.

[0285] In the above formula (Ic), q is an integer between 0 and 4, preferably between 0 and 2, and more preferably 0 or 1. Here, "q is 0" means R 3c This means that it does not exist.

[0286] q R in equation (Ic) 3c Each of these is independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. One embodiment of the manufacturing method of the present invention, q R 3c Each of these may be independently and preferably substituted with C 1-6 Alkyl alkyl groups, or substituted C 1-6 The group is an alkoxy group or an electron-withdrawing group, more preferably a halogen atom or a nitro group, and even more preferably a nitro group. Another embodiment of the manufacturing method of the present invention, q R 3c Each of these is independently preferably an electron-withdrawing group, more preferably a halogen atom, C 1-6 It is a perfluoroalkyl group or a nitro group, and more preferably C1-6 The group is a perfluoroalkyl group or a nitro group, and is particularly preferably a trifluoromethyl group.

[0287] Suitable compounds (Ic) include the following. Each of these compounds may be used individually or in combination of two or more.

[0288] (Compound (Ic-i)) In the above equation (Ic), R 1c but, (1) A C group which may be substituted with one or two substituents selected from the group consisting of a substituted phenyl group and a substituted 5 or 6-membered monocyclic heteroaryl group. 1-6 alkyl group, (2) A phenyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (3) A 5 or 6-membered monocyclic heteroaryl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. And, Y 1c However, these are -S(=O)-, -S(=O)2-, -C(=O)-, or -C(=S)-, p is 0 or 1, R 2c However, it is a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. Ring C is a 6-membered aromatic hydrocarbon ring or a 6-membered nitrogen-containing aromatic heterocycle. q is an integer from 0 to 4, and q R 3c However, each is independently a potentially substituted alkyl group, a potentially substituted alkoxy group, or an electron-withdrawing group. A compound (referred to as "compound (Ic-i)" in this specification).

[0289] (Compound (Ic-ii)) In the above equation (Ic), R 1c but, (1) A C group which may be substituted with one or two selected substituents selected from the group consisting of optionally substituted phenyl groups and optionally substituted 5 or 6-membered monocyclic heteroaryl groups. 1-6 alkyl group, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A phenyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. And, Y 1c However, -S(=O)-, -S(=O)2-, -C(=O)- or -C(=S)- p is 0 or 1, R 2c However, hydrogen atoms may be substituted C 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group or an electron-withdrawing group. Ring C is a 6-membered aromatic hydrocarbon ring or a 6-membered nitrogen-containing aromatic heterocycle. q is an integer between 0 and 2, and q R 3c However, each can be substituted independently of the others. 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group or an electron-withdrawing group. Compound (referred to as "compound (Ic-ii)" in this specification).

[0290] (Compound (Ic-iii)) In the above equation (Ic), R 1c but, (1) C which may be substituted with one or two pyridyl groups 1-6 alkyl group, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A phenyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups, or (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups. And, Y 1c However, -C(=O)- p is 0, Ring C is a benzene ring or a pyridine ring, q is an integer between 0 and 2, q R 3c However, each is independently a halogen atom or a nitro group. Compounds (referred to as "compound (Ic-iii)" in this specification).

[0291] (Compound (Ic-iv)) In the above equation (Ic), R 1c but, (1) C which may be substituted with one or two pyridyl groups 1-6 alkyl group, (2) One or two C 1-6 A phenyl group which may be substituted with an alkyl group, or (3) Pyridyl group (especially 2-pyridyl group or 4-pyridyl group) And, Y 1c However, -C(=O)- p is 0, Ring C is a benzene ring or a pyridine ring, q is an integer between 0 and 2, and q R 3c However, it is a nitro group. Compounds (referred to as "compound (Ic-iv)" herein).

[0292] (Compound (Ic-i')) In the above equation (Ic), R 1c but, (1) A C group which may be substituted with one or two substituents selected from the group consisting of a substituted phenyl group and a substituted 5 or 6-membered monocyclic heteroaryl group. 1-6 alkyl group, (2) A phenyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (3) A 5 or 6-membered monocyclic heteroaryl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. And, Y 1c However, these are -S(=O)-, -S(=O)2-, -C(=O)-, or -C(=S)-, p is 0 or 1, R 2c However, it is a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. Ring C is a 6-membered aromatic hydrocarbon ring, a 6-membered nitrogen-containing aromatic heterocycle, or a 10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom. q is an integer from 0 to 4, and q R 3c However, each is independently a potentially substituted alkyl group, a potentially substituted alkoxy group, or an electron-withdrawing group. A compound (referred to as "compound (Ic-i')" in this specification).

[0293] (Compound (Ic-ii')) In the above equation (Ic), R 1c but, (1) A C group which may be substituted with one or two selected substituents selected from the group consisting of optionally substituted phenyl groups and optionally substituted 5 or 6-membered monocyclic heteroaryl groups. 1-6 alkyl group, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A phenyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. And, Y 1c However, -S(=O)-, -S(=O)2-, -C(=O)- or -C(=S)- p is 0 or 1, R 2c However, hydrogen atoms may be substituted C 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group or an electron-withdrawing group. Ring C is a 6-membered aromatic hydrocarbon ring, a 6-membered nitrogen-containing aromatic heterocycle, or a 10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom. q is an integer between 0 and 2, and q R 3c However, each can be substituted independently of the others. 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group or an electron-withdrawing group. A compound (referred to as "compound (Ic-ii')" in this specification).

[0294] (Compound (Ic-iii')) In the above equation (Ic), R 1c but, (1)C 1-6 alkyl group, (2) C may be substituted1-6 Alkyl alkyl groups, or substituted C 1-6 A phenyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups, or (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups. And, Y 1c However, -C(=O)- p is 0 or 1, Ring C is a benzene ring, a pyridine ring, or a quinoline ring. q is an integer between 0 and 2, q R 3c However, each is independent of the halogen atom, C 1-6 It is a perfluoroalkyl group or a nitro group. A compound (referred to as "compound (Ic-iii')" herein).

[0295] (Compound (Ic-iv')) In the above equation (Ic), R 1c but, (1)C 1-6 alkyl group, (2)C 1-6 Alkyl alkyl group, C 1-6 A phenyl group which may be substituted with one or two substituents selected from the group consisting of alkoxy groups and halogen atoms, (3) One or two C 1-6 Pyridyl groups substituted with alkyl groups (e.g., 4-methyl-2-pyridyl group) And, Y 1c However, -C(=O)- p is 0 or 1, Ring C is a benzene ring, a pyridine ring, or a quinoline ring. q is an integer between 0 and 2, and q R 3c However, C1-6 It is a perfluoroalkyl group. A compound (referred to as "compound (Ic-iv')" herein).

[0296] (Preferred specific examples of compound (Ic)) Preferred specific examples of compound (Ic) include compounds represented by any of the following formulas (Ic-1) to (Ic-17).

[0297] [ka]

[0298] [ka]

[0299] In one embodiment of the production method of the present invention, compound (Ic) is preferably 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)), 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-2)), 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one (compound (Ic-3)), 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-4)), 5-nitro-2-phenyl-1, 2-Benzothiazol-3(2H)-one (compound (Ic-5)), 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazol-3(2H)-one (compound (Ic-6)), 5-nitro-2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one (compound (Ic-7)), 5-nitro-2-(4-pyridyl)-1,2-benzothiazol-3(2H)-one (compound (Ic-8)), 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazo 2-phenylisothiazolo[4,5-c]pyridine-3(2H)-one (compound (Ic-9)), 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one (compound (Ic-11)), 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-12)), 2-phenyl-2H-1,2-benzothiadin-3(4H)-one (compound (Ic-13)), 2-(2,6-dimethylphenyl It is at least one selected from the group consisting of isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-14)), 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-15)), 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-16)), and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-17)).

[0300] In another embodiment of the production method of the present invention, compound (Ic) is preferably 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)), 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-2)), 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one (compound (Ic-3)), 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-4)), 5-nitro-2-phenyl-1,2-benzothiazole-3(2H)-one (compound At least one selected from the group consisting of (Ic-5), 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazole-3(2H)-one (compound (Ic-6)), 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-7)), 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-8)), and 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3(2H)-one (compound (Ic-9)).

[0301] In another embodiment of the manufacturing method of the present invention, compound (Ic) is preferably 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)), 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-2)), 5-nitro-2-phenyl-1,2-benzothiazole-3(2H)-one (compound (Ic-5)), 2-phenylisothiazolo[4,5-c]pyridine-3(2H)-one (compound (Ic-10)), 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one (compound (Ic-11)), 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H) It is at least one selected from the group consisting of -one (compound (Ic-12)), 2-phenyl-2H-1,2-benzothiazine-3(4H)-one (compound (Ic-13)), 2-(2,6-dimethylphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-14)), 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-15)), 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-16)), and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-17)).

[0302] In another embodiment of the manufacturing method of the present invention, compound (Ic) is preferably 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)), 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-2)), 2-phenylisothiazolo[4,5-c]pyridine-3(2H)-one (compound (Ic-10)), 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one (compound (Ic-11)), 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-12)), 2-phenyl- It is at least one selected from the group consisting of 2H-1,2-benzothiazin-3(4H)-one (compound (Ic-13)), 2-(2,6-dimethylphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-14)), 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-15)), 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-16)), and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-17)).

[0303] In the production method of the present invention, compound (Ic) is most preferably 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)).

[0304] 2-1-1B. Compound (IB) Next, compound (IB) will be described. In compound (IB), R in formula (I) above 1 is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, preferably an optionally substituted C 1-10 The group is an alkyl group, an optionally substituted phenyl group, an optionally substituted 5- or 6-membered monocyclic heteroaryl group, or an optionally substituted 8- to 14-membered fused polycyclic heteroaryl group.

[0305] In compound (IB), R in formula (I) 2 This is an optionally substituted aryl group or an optionally substituted heteroaryl group, preferably an optionally substituted phenyl group, an optionally substituted 5 or 6-membered monocyclic heteroaryl group, or an optionally substituted 8 to 14-membered condensed polycyclic heteroaryl group.

[0306] 2-1-1B-1. Compound (Ib) Compound (IB) is preferably a compound represented by the following formula (Ib) (excluding 2,2-dipyridyl disulfide). Compound (Ib) may be used alone or in combination of two or more types.

[0307] [ka]

[0308] The following describes each group of compound (Ib). R in equation (Ib) above 1b is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, preferably an optionally substituted C 1-10 The group is an alkyl group, an optionally substituted phenyl group, an optionally substituted 5- or 6-membered monocyclic heteroaryl group, or an optionally substituted 8- to 14-membered fused polycyclic heteroaryl group.

[0309] In one embodiment of the manufacturing method of the present invention (hereinafter referred to as "embodiment (A)"), R 1b is more (1) an alkyl group substituted with one carboxyl group, (2) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (6) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (7) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (8) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (9) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (10) A phenyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (11) A 2-imidazolyl group which may be substituted with 1 to 3 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. That is the case.

[0310] In embodiment (A), R 1b More preferably (1) an alkyl group substituted with one carboxyl group, (2) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (6) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (7) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (8) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (9) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. That is the case.

[0311] In embodiment (A), R 1b Particularly preferred (1) C substituted with one carboxyl group 1-6 alkyl group, (2) C may be substituted 1-6Alkyl alkyl groups, or substituted C 1-6 A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (4) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (5) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (6) C may not be substituted 1-6 A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of alkyl groups and optionally substituted phenyl groups, (7) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 The (pyridine-N-oxide)-2-yl group may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. (8) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (9) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. That is the case.

[0312] In embodiment (A), R 1b Most preferably, C is substituted with one carboxyl group. 1-6 These are alkyl groups, 2-benzothiazolyl groups, 2-benzimidazolyl groups, 2-benzoxazolyl groups, 5-(1,2,3-triazolyl) groups, 1-phenyl-1H-tetrazole-5-yl groups, (pyridine-N-oxide)-2-yl groups, 4-pyridyl groups, or 2-pyridyl groups.

[0313] In another embodiment of the manufacturing method of the present invention (hereinafter referred to as "embodiment (B)"), R 1b is more (1) an alkyl group substituted with one carboxyl group, (2) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (6) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (7) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (8) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. That is the case.

[0314] In embodiment (B), R 1b More preferably (1) C substituted with one carboxyl group 1-6 alkyl group, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (4) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (5) C may be substituted 1-6 A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of alkyl groups and optionally substituted phenyl groups, (6) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 The (pyridine-N-oxide)-2-yl group may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. (7) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (8) C may be substituted1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. That is the case.

[0315] In embodiment (B), R 1b Particularly preferred is a C substituted with one carboxyl group. 1-6 These are alkyl groups, 2-benzothiazolyl groups, 2-benzimidazolyl groups, 2-benzoxazolyl groups, 1-phenyl-1H-tetrazole-5-yl groups, (pyridine-N-oxide)-2-yl groups, 4-pyridyl groups, or 2-pyridyl groups.

[0316] In formula (Ib), ring B is a 6-membered aromatic hydrocarbon ring (i.e., a benzene ring) or a 5- or 6-membered aromatic heterocycle, preferably a 5- or 6-membered aromatic heterocycle, more preferably a triazole ring, a tetrazole ring, a pyridine ring, or a pyridine-N-oxide ring, and even more preferably a tetrazole ring, a pyridine ring, or a pyridine-N-oxide ring.

[0317] In equation (Ib) above, n is an integer from 0 to 5, and n R 2b Each of these is independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, more preferably an optionally substituted C 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group or an electron-withdrawing group. Here, "n is 0" means R 2b This means that it does not exist.

[0318] If n in equation (Ib) is an integer greater than or equal to 2, then two adjacent R 2b Together with ring B, it may form a substituted or alternatively substituted bicyclic fused aromatic heterocycle (preferably a substituted or alternatively substituted benzimidazole ring, a substituted or alternatively substituted benzoxazole ring, or a substituted or alternatively substituted benzthiazole ring).

[0319] In one embodiment of the manufacturing method of the present invention (hereinafter referred to as "embodiment (C)"), the formula (r1) of compound (Ib):

[0320] [ka]

[0321] The group represented is preferably (1) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (2) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (6) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (7) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (8) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (9) A phenyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (10) A 2-imidazolyl group which may be substituted with 1 to 3 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. That is the case.

[0322] In embodiment (C), the base (r1) is more preferably (1) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (2) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (6) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (7) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (8) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. That is the case.

[0323] In embodiment (C), the base (r1) is more preferably (1) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (4) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (5) C may be substituted 1-6 A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of alkyl groups and optionally substituted phenyl groups, (6) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6The (pyridine-N-oxide)-2-yl group may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. (7) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (8) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. That is the case.

[0324] In embodiment (C), the group (r1) is particularly preferably a 2-benzothiazolyl group, a 2-benzimidazolyl group, a 2-benzoxazolyl group, a 5-(1,2,3-triazolyl) group, a 1-phenyl-1H-tetrazole-5-yl group, a (pyridine-N-oxide)-2-yl group, a 4-pyridyl group, or a 2-pyridyl group, and most preferably a 2-benzothiazolyl group, a 2-benzimidazolyl group, a 2-benzoxazolyl group, a 5-(1,2,3-triazolyl) group, a 1-phenyl-1H-tetrazole-5-yl group, or a (pyridine-N-oxide)-2-yl group.

[0325] In another embodiment of the manufacturing method of the present invention (hereinafter referred to as "embodiment (D)"), the base (r1) is preferably (1) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (2) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (5) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (6) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (7) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. That is the case.

[0326] In embodiment (D), the base (r1) is more preferably (1) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (4) C may be substituted 1-6A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of alkyl groups and optionally substituted phenyl groups, (5) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 The (pyridine-N-oxide)-2-yl group may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. (6) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (7) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. That is the case.

[0327] In embodiment (D), the group (r1) is particularly preferably a 2-benzothiazolyl group, a 2-benzimidazolyl group, a 2-benzoxazolyl group, a 1-phenyl-1H-tetrazole-5-yl group, a (pyridine-N-oxide)-2-yl group, a 4-pyridyl group, or a 2-pyridyl group, and most preferably a 2-benzothiazolyl group, a 2-benzimidazolyl group, a 2-benzoxazolyl group, a 1-phenyl-1H-tetrazole-5-yl group, or a (pyridine-N-oxide)-2-yl group.

[0328] Suitable compounds (Ib) include the following. Each of these compounds may be used individually or in combination of two or more.

[0329] (Compound (Ib-i)) In the above equation (Ib), R 1b but, (1) an alkyl group substituted with one carboxyl group, (2) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (6) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (7) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (8) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (9) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (10) A phenyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (11) A 2-imidazolyl group which may be substituted with 1 to 3 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. and The base (r1) is (1) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (2) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (6) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (7) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (8) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (9) A phenyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (10) A 2-imidazolyl group which may be substituted with 1 to 3 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. A compound that is (referred to as "compound (Ib-i)" in this specification).

[0330] (Compound (Ib-ii)) In the above equation (Ib), R 1b but, (1) an alkyl group substituted with one carboxyl group, (2) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (6) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (7) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (8) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (9) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. and The base (r1) is (1) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (2) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (6) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (7) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (8) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. A compound that is (referred to as "compound (Ib-ii)" in this specification).

[0331] (Compound (Ib-iii)) In the above equation (Ib), R 1b but, (1) C substituted with one carboxyl group 1-6 alkyl group, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (4) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (5) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (6) C may not be substituted 1-6 A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of alkyl groups and optionally substituted phenyl groups, (7) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 The (pyridine-N-oxide)-2-yl group may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. (8) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (9) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. and The base (r1) is (1) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (4) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (5) C may be substituted 1-6 A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of alkyl groups and optionally substituted phenyl groups, (6) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 The (pyridine-N-oxide)-2-yl group may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. (7) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (8) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. A compound that is (referred to as "compound (Ib-iii)" in this specification).

[0332] (Compound (Ib-iv)) In the above equation (Ib), R 1b However, C substituted with one carboxyl group 1-6 Alkyl group, 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 5-(1,2,3-triazolyl) group, 1-phenyl-1H-tetrazole-5-yl group, (pyridine-N-oxide)-2-yl group, 4-pyridyl group, or 2-pyridyl group, and Compounds in which group (r1) is a 2-benzothiazolyl group, a 2-benzimidazolyl group, a 2-benzoxazolyl group, a 5-(1,2,3-triazolyl) group, a 1-phenyl-1H-tetrazole-5-yl group, a (pyridine-N-oxide)-2-yl group, a 4-pyridyl group, or a 2-pyridyl group (referred to as "compounds (Ib-iv)" herein).

[0333] (Compound (Ib-v)) In the above equation (Ib), R 1b However, C substituted with one carboxyl group 1-6 Alkyl group, 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 5-(1,2,3-triazolyl) group, 1-phenyl-1H-tetrazole-5-yl group, (pyridine-N-oxide)-2-yl group, 4-pyridyl group, or 2-pyridyl group, and The compound (sometimes referred to as "compound (Ib-v)" herein) is a compound in which group (r1) is a 2-benzothiazolyl group, a 2-benzimidazolyl group, a 2-benzoxazolyl group, a 5-(1,2,3-triazolyl) group, a 1-phenyl-1H-tetrazole-5-yl group, or a (pyridine-N-oxide)-2-yl group.

[0334] (Compound (Ib-i')) In the above equation (Ib), R 1b but, (1) an alkyl group substituted with one carboxyl group, (2) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (6) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (7) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (8) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, and The base (r1) is (1) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (2) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (4) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (5) A (pyridine-N-oxide)-2-yl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (6) A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (7) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, A compound that is (referred to herein as "compound (Ib-i')").

[0335] (Compound (Ib-ii')) In the above equation (Ib), R 1b but, (1) C substituted with one carboxyl group 1-6 alkyl group, (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (4) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (5) C may be substituted 1-6 A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of alkyl groups and optionally substituted phenyl groups, (6) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 The (pyridine-N-oxide)-2-yl group may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. (7) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (8) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. and The base (r1) is (1) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (2) C may be substituted1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (3) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, (4) C may be substituted 1-6 A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of alkyl groups and optionally substituted phenyl groups, (5) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 The (pyridine-N-oxide)-2-yl group may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. (6) C may not be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 4-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups, or (7) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups and electron-withdrawing groups. A compound that is (referred to herein as "compound (Ib-ii')").

[0336] (Compound (Ib-iii')) In the above equation (Ib), R 1b However, C substituted with one carboxyl group 1-6Alkyl group, 2-benzothiazolyl group, 2-benzoimidazolyl group, 2-benzoxazolyl group, 1-phenyl-1H-tetrazole-5-yl group, (pyridine-N-oxide)-2-yl group, 4-pyridyl group, or 2-pyridyl group, and Compounds in which group (r1) is a 2-benzothiazolyl group, a 2-benzimidazolyl group, a 2-benzoxazolyl group, a 1-phenyl-1H-tetrazole-5-yl group, a (pyridine-N-oxide)-2-yl group, a 4-pyridyl group, or a 2-pyridyl group (referred to herein as "compound (Ib-iii')").

[0337] (Compound (Ib-iv')) In the above equation (Ib), R 1b However, C substituted with one carboxyl group 1-6 Alkyl group, 2-benzothiazolyl group, 2-benzoimidazolyl group, 2-benzoxazolyl group, 1-phenyl-1H-tetrazole-5-yl group, (pyridine-N-oxide)-2-yl group, 4-pyridyl group, or 2-pyridyl group, and The compound (which may be referred to as "compound (Ib-iv')" herein) is a compound in which group (r1) is a 2-benzothiazolyl group, a 2-benzimidazolyl group, a 2-benzoxazolyl group, a 1-phenyl-1H-tetrazole-5-yl group, or a (pyridine-N-oxide)-2-yl group.

[0338] (Preferred specific examples of compound (Ib)) Preferred specific examples of compound (Ib) include compounds represented by any of the following formulas (Ib-1) to (Ib-12) (wherein Ph in formula (Ib-7) represents a phenyl group).

[0339] [ka]

[0340] In one embodiment of the production method of the present invention, compound (Ib) is preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)), 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)), 2,2'-dibenzimidazolyl disulfide (compound (Ib-4)), 2,2'-dibenzoxazolyl disulfide (compound (Ib-5)), 5,5'-di(1,2,3-triazolyl) disulfide (compound (Ib-6)). It is at least one selected from the group consisting of ), 5,5'-dithiobis(1-phenyl-1H-tetrazole) (compound (Ib-7)), 2,2'-dithiobis(pyridine-N-oxide) (compound (Ib-8)), 4,4'-dipyridyl disulfide (compound (Ib-9)), 3-(2-pyridyldithio)propanoic acid (compound (Ib-10)), 2,2'-dithiobis(1H-imidazole) (compound (Ib-11)), and 2,2'-dithiobis(1-methyl-1H-imidazole) (compound (Ib-12)).

[0341] In another embodiment of the manufacturing method of the present invention, compound (Ib) is preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)), 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)), 2,2'-dibenzoimidazolyl disulfide (compound (Ib-4)), 2,2'-dibenzoxazolyl disulfide (compound (Ib-5)) It is at least one selected from the group consisting of ), 5,5'-di(1,2,3-triazolyl) disulfide (compound (Ib-6)), 5,5'-dithiobis(1-phenyl-1H-tetrazole) (compound (Ib-7)), 2,2'-dithiobis(pyridine-N-oxide) (compound (Ib-8)), 4,4'-dipyridyl disulfide (compound (Ib-9)), and 3-(2-pyridyldithio)propanoic acid (compound (Ib-10)).

[0342] In another embodiment of the manufacturing method of the present invention, compound (Ib) is preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)), 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)), 2,2'-dibenzoimidazolyl disulfide (compound (Ib-4)), or 2,2'-dibenzox It is at least one selected from the group consisting of zolyl disulfide (compound (Ib-5)), 5,5'-dithiobis(1-phenyl-1H-tetrazole) (compound (Ib-7)), 2,2'-dithiobis(pyridine-N-oxide) (compound (Ib-8)), 4,4'-dipyridyl disulfide (compound (Ib-9)), and 3-(2-pyridyldithio)propanoic acid (compound (Ib-10)).

[0343] In another embodiment of the manufacturing method of the present invention, compound (Ib) is preferably at least one selected from the group consisting of 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)), 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)), 2,2'-dibenzimidazolyl disulfide (compound (Ib-4)), 2,2'-dibenzoxazolyl disulfide (compound (Ib-5)), 5,5'-di(1,2,3-triazolyl) disulfide (compound (Ib-6)), 5,5'-dithiobis(1-phenyl-1H-tetrazole) (compound (Ib-7)), and 2,2'-dithiobis(pyridine-N-oxide) (compound (Ib-8)).

[0344] In another embodiment of the manufacturing method of the present invention, compound (Ib) is preferably at least one selected from the group consisting of 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)), 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)), 2,2'-dibenzimidazolyl disulfide (compound (Ib-4)), 2,2'-dibenzoxazolyl disulfide (compound (Ib-5)), 5,5'-dithiobis(1-phenyl-1H-tetrazole) (compound (Ib-7)), and 2,2'-dithiobis(pyridine-N-oxide) (compound (Ib-8)).

[0345] In the manufacturing method of the present invention, compound (Ib) is most preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)).

[0346] 2-1-2. Compound (ID) ~ Compound (IG) Compounds other than those mentioned above (IA) and (IB), namely compound (I), can be classified as follows: In equation (I) above, X 1 However, it is a single bond, R 1 However, is a halogen atom, and R in formula (I) 2 This refers to a compound (referred to herein as "compound (ID)") which is an optionally substituted aryl group or an optionally substituted heteroaryl group. In equation (I) above, X 1 However, -S(=O)2- and R 1 However, it is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 This refers to a compound (referred to herein as "compound (IE)") which is an optionally substituted aryl group or an optionally substituted heteroaryl group. In equation (I) above, X 1 However, it is an oxygen atom, and R 1 and R 2However, together with the sulfur and oxygen atoms to which they are bonded, compounds (referred to herein as "compound (IF)") may form substituted heterocycles, and In equation (I) above, X 1 -N(-R 3 )- and R 1 and R 3 However, together with the nitrogen atoms to which they are bonded, they form a heterocycle which may be substituted, and R 2 However, the compound is an optionally substituted aryl group or an optionally substituted heteroaryl group (hereinafter referred to as "compound (IG)").

[0347] Compounds (ID), (IE), (IF), and (IG) may be used individually or in combination of two or more. Compounds (ID), (IE), (IF), and (IG) will be described in order below.

[0348] 2-1-2D. Compound (ID) X in equation (I) above 1 This section describes compounds (ID) in which the bond is a single bond. Compound (ID) is represented by the following formula (Id): R 2d -SR 1d (Id) [In the formula, R 1d R indicates a halogen atom, and 2d This indicates an optionally substituted aryl group or an optionally substituted heteroaryl group. It can be expressed as follows. Compound (Id) (= Compound (ID)) may be used alone or in combination of two or more types.

[0349] R in the above formula (Id) 1d R is preferably a chlorine atom. 2d Preferably, is a heteroaryl group which may be substituted, more preferably a 5 or 6-membered monocyclic heteroaryl group which may be substituted with an electron-withdrawing group, and even more preferably a pyridyl group which may be substituted with a nitro group.

[0350] Suitable compounds (Id) include the following. Each of these compounds may be used individually or in combination of two or more.

[0351] (Compound (Id-i)) In the above formula (Id), R 1d However, it is a halogen atom, and R 2d However, the compound is a heteroaryl group which may be substituted (referred to herein as "compound (Id-i)").

[0352] (Compound (Id-ii)) In the above formula (Id), R 1d However, it is a chlorine atom, and R 2d However, the compound is a 5- or 6-membered monocyclic heteroaryl group which may be substituted with an electron-withdrawing group (hereinafter referred to as "compound (Id-ii)").

[0353] (Compound (Id-iii)) In the above formula (Id), R 1d However, it is a chlorine atom, and R 2d However, a compound that is a pyridyl group which may be substituted with a nitro group (referred to herein as "compound (Id-iii)").

[0354] (Preferred specific examples of compound (Id)) A preferred specific example of compound (Id) is 3-nitro-2-pyridinesulfenyl chloride, represented by the following formula (Id-1).

[0355] [ka]

[0356] 2-1-2E. Compound (IE) X in equation (I) above 1 This section describes compounds (IE) whose integral is -S(=O)2-. Compounds (IE) are given by the following formula (Ie):

[0357] [ka]

[0358] [In the formula, R 1e and R 2e Each of these independently represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. It can be expressed as follows. Compound (Ie) (= compound (IE)) may be used alone or in combination of two or more types.

[0359] R in the above equation (Ie) 1e and R 2e Each of these is independently preferably a substituted aryl group, and more preferably a substituted phenyl group.

[0360] (Preferred specific examples of compound (Ie)) A preferred specific example of compound (Ie) is (S)-phenyl, represented by the following formula (Ie-1). It is a benzenethiosulfonate.

[0361] [ka]

[0362] 2-1-2F. Compound (IF) In equation (I) above, X 1 However, it is an oxygen atom, and R 1 and R 2 However, we will describe compounds (IF) that form a heterocycle, which may be substituted, together with the sulfur and oxygen atoms to which they are bonded. Compound (IF) is preferably of the following formula (If):

[0363] [ka]

[0364] [In the formula, Ring D represents a 6-membered aromatic hydrocarbon ring or a 5- or 6-membered aromatic heterocycle. w represents an integer from 0 to 4. w pieces of R 1f Each of these independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. X 1f -C(=O)-, -C(=S)-, -S(=O)2-, or -CHR 2f - indicates R 2f This represents a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and v represents an integer between 0 and 3. This is a compound represented by (If). Compound (If) may be used alone or in combination of two or more types.

[0365] In the above formula (If), ring D is preferably a 6-membered aromatic hydrocarbon ring (i.e., a benzene ring).

[0366] In the above formula (If), w is preferably an integer between 0 and 2, and more preferably 0 or 1. Here, "w is 0" means that R 1f This means that it does not exist.

[0367] w Rs in the above equation (If) 1f Each of these may be independently and preferably substituted with C 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group, a halogen atom, or a nitro group.

[0368] R in the above equation (If) 2f Preferably a hydrogen atom, and possibly a substituted C 1-6 Alkyl alkyl groups, or substituted C 1-6 An alkoxy group or an electron-withdrawing group, more preferably a hydrogen atom, which may be substituted with C 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group, a halogen atom, or a nitro group. X in the above equation (If) 1f Preferably, it is -C(=O)-.

[0369] In the above formula (If), v is preferably 0. Here, "v is 0" means that the -CH2- in parentheses in the above formula (If) does not exist.

[0370] Suitable compounds (If) include the following. Each of these compounds may be used individually or in combination of two or more.

[0371] (Compound (If-i)) In the above expression (If), Ring D is a benzene ring, w is preferably an integer between 0 and 2, and w pieces of R 1f However, each is independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. X 1f However, -C(=O)- and A compound in which v is an integer between 0 and 3 (referred to herein as "compound (If-i)").

[0372] (Compound (If-ii)) In the above expression (If), Ring D is a benzene ring, w is preferably an integer between 0 and 2, and w pieces of R 1f However, each can be substituted independently of the others. 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group, a halogen atom, or a nitro group. X 1f However, -C(=O)- and A compound in which v is an integer between 0 and 3 (referred to herein as "compound (If-ii)").

[0373] (Compound (If-iii)) In the above expression (If), Ring D is a benzene ring, w is preferably an integer of 0 or 1, R 1f is an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom or a nitro group, and X 1f is -C(=O)-, a compound in which v is 0 (described herein as "compound (If-iii)").

[0374] (Preferred specific examples of compound (If)) Preferred specific examples of compound (If) are 3H-2,1-benzoxathiol-3-one represented by the following formula (If-1).

[0375]

Chemical formula

[0376] 2-1-2G. Compound (IG) For the compound (IG) in which X 1 in the above formula (I) is -N(-R 3 ​​​​​​​​​​​​​​​​​​​​​​​​​​Each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, an oxo group, or an electron-withdrawing group, When x is an integer of 2 or more, two adjacent Rs 1g may together with ring E form an optionally substituted bicyclic fused nitrogen-containing heterocycle, and R 2g represents an optionally substituted aryl group or an optionally substituted heteroaryl group.] It is a compound represented by. Compound (Ig) may be used alone or in combination of two or more.

[0379] R in the above formula (Ig) 2g is preferably an optionally substituted heteroaryl group, more preferably an optionally substituted 5- or 6-membered monocyclic nitrogen-containing heteroaryl group, or an optionally substituted 8- to 14-membered fused polycyclic nitrogen-containing heteroaryl group.

[0380] R 2g is even more preferably (1) A 2-pyridyl group optionally substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group, and an electron-withdrawing group, or (2) A 2-benzothiazolyl group optionally substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group, and an electron-withdrawing group is.

[0381] R 2g is particularly preferably (1) A 2-pyridyl group optionally substituted with I to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom, and a nitro group, or (2) A 2-pyridyl group optionally substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups. That is the case.

[0382] R 2g The most preferred group is a 2-pyridyl group or a 2-benzothiazolyl group.

[0383] In the above formula (Ig), x is an integer between 0 and 6, preferably between 2 and 6. Here, "x is 0" means R 1g This means that it does not exist.

[0384] x R values ​​in the above equation (Ig) 1g Each of these may be independently and preferably substituted with C 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group, an oxo group, a halogen atom, or a nitro group.

[0385] Formula (r17) in compound (Ig):

[0386] [ka]

[0387] The base represented by formula (r17') is preferably:

[0388] [ka]

[0389] [In the formula, Ring F is a 4- to 8-membered nitrogen-containing heterocycle. y represents an integer from 0 to 4, and y R 3g Each of these independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. If y is an integer greater than or equal to 2, then two adjacent R 3gIt may form a substituted, bicyclic condensed nitrogen-containing heterocycle together with ring F. It is a base represented by .

[0390] In the above equation (r17'), y is between 0 and 4. Here, "y is 0" means R 3g This means that it does not exist.

[0391] The y R in the above equation (r17') 3g Each of these can be substituted independently, preferably C 1-6 Alkyl alkyl groups, or substituted C 1-6 An alkoxy group, a halogen atom, or a nitro group, or two adjacent R groups 3g Preferably, together with ring F, it forms a substituted or otherwise substituted bicyclic condensed nitrogen-containing heterocycle. The bicyclic condensed nitrogen-containing heterocycle is preferably an isoindole ring which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, and more preferably an optionally substituted C 1-6 Alkyl alkyl groups, or substituted C 1-6 The isoindole ring may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups.

[0392] Ring F is preferably a pyrrolidine ring or two adjacent R rings. 3g Along with this, it forms an isoindole ring which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups.

[0393] Ring F is more preferably a pyrrolidine ring, or two adjacent R 3g Along with, C may still be substituted. 1-6 Alkyl alkyl groups, or substituted C 1-6 It forms an isoindole ring which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups.

[0394] Compound (Ig) is preferably represented by the following formula (Ig'), with an imide bond (-C(=O)-N(-SR 2g It is a compound containing )-C(=O)-) (the definition of the symbols in the following formula is as described above).

[0395] [ka]

[0396] Suitable compounds (Ig') include the following. Each of these compounds may be used individually or in combination of two or more.

[0397] (Compound (Ig-ia)) In the above formula (Ig'), Ring F is a pyrrolidine ring, y is an integer between 0 and 4, y R 3g However, each is independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. R 2g However, the compound is a heteroaryl group which may be substituted (referred to herein as "compound (Ig-ia)").

[0398] (Compound (Ig-ib)) In the above formula (Ig'), Ring F is adjacent to two R 3g Together, it forms an isoindole ring which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, and R 2g However, the compound is a heteroaryl group that may be substituted (referred to herein as "compound (Ig-ib)").

[0399] (Compound (Ig-ii-a)) In the above formula (Ig'), Ring F is a pyrrolidine ring, y is an integer between 0 and 4, y R 3g However, each is independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and R 2g However, the compound is a substituted 5 or 6-membered monocyclic nitrogen-containing heteroaryl group, or a substituted 8- to 14-membered condensed polycyclic nitrogen-containing heteroaryl group (referred to herein as "compound (Ig-ii-a)").

[0400] (Compound (Ig-ii-b)) In the above formula (Ig'), Ring F is adjacent to two R 3g Together, it forms an isoindole ring which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, and R 2g However, the compound is a substituted 5- or 6-membered monocyclic nitrogen-containing heteroaryl group, or a substituted 8- to 14-membered condensed polycyclic nitrogen-containing heteroaryl group (referred to herein as "compound (Ig-ii-b)").

[0401] (Compound (Ig-iii-a)) In the above formula (Ig'), Ring F is a pyrrolidine ring, y is an integer between 0 and 4, y R 3g However, each is independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and R 2g but, (1) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (2) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. A compound that is (referred to herein as "compound (Ig-iii-a)").

[0402] (Compound (Ig-iii-b)) In the above formula (Ig'), Ring F is adjacent to two R 3g Together, it forms an isoindole ring which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, and R 2g but, (1) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (2) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. A compound that is (referred to herein as "compound (Ig-iii-b)").

[0403] (Compound (Ig-iv-a)) In the above formula (Ig'), Ring F is a pyrrolidine ring, y is an integer between 0 and 4, y R 3g However, even if they are substituted independently, C 1-6 Alkyl alkyl groups, or substituted C 1-6 It is an alkoxy group, a halogen atom, or a nitro group. R 2g but, (1) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups, or (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups. A compound that is (referred to herein as "compound (Ig-iv-a)").

[0404] (Compound (Ig-iv-b)) In the above formula (Ig'), Ring F is adjacent to two R 3g Along with, C may still be substituted. 1-6 Alkyl alkyl groups, or substituted C 1-6 An isoindole ring is formed which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups, and R 2g but, (1) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups, or (2) C may be substituted 1-6 Alkyl alkyl groups, or substituted C 1-6 A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of alkoxy groups, halogen atoms, and nitro groups. A compound that is (referred to herein as "compound (Ig-iv-b)").

[0405] (Preferred specific examples of compound (Ig)) Preferred specific examples of compound (Ig) include 1-(2-pyridinylthio)-2,5-pyrrolidinedione represented by the following formula (Ig-1), 1-(2-benzothiazolylthio)-2,5-pyrrolidinedione represented by the following formula (Ig-2), 2-(2-pyridinylthio)-1H-isoindole-1,3(2H)-dione represented by the following formula (Ig-3), and 2-(2-benzothiazolylthio)-1H-isoindole-1,3(2H)-dione represented by the following formula (Ig-4).

[0406] [ka]

[0407] 2-1-3. Preferred Compounds (I) In the manufacturing method of the present invention, compound (I) is preferably compound (Ia) and / or compound (Ib). Among compound (Ia), compound (Ic) is preferred.

[0408] In one embodiment of the manufacturing method of the present invention, compound (I) is preferably compound (Ib) and / or compound (Ic), more preferably compound (Ib-i) and / or compound (Ic-i), even more preferably compound (Ib-ii) and / or compound (Ic-ii), even more preferably compound (Ib-iii) and / or compound (Ic-iii), even more preferably compound (Ib-iv) and / or compound (Ic-iv), and particularly preferably compound (Ib-v) and / or compound (Ic-iv).

[0409] In another embodiment of the manufacturing method of the present invention, compound (I) is preferably compound (Ib-i') and / or compound (Ic-i'), more preferably compound (Ib-ii') and / or compound (Ic-ii'), even more preferably compound (Ib-iii') and / or compound (Ic-iii'), and still more preferably compound (Ib-iv') and / or compound (Ic-iv').

[0410] In another embodiment of the manufacturing method of the present invention, compound (I) is preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)), 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)), 2,2'-dibenzimidazolyl disulfide (compound (Ib-4)), 2,2'-dibenzoxazolyl disulfide (compound (Ib-5)), 5,5'-di(1,2,3-triazolyl) disulfide (compound (Ib-1) (Ib-6)), 5,5'-dithiobis(1-phenyl-1H-tetrazole) (compound (Ib-7)), 2,2'-dithiobis(pyridine-N-oxide) (compound (Ib-8)), 4,4'-dipyridyl disulfide (compound (Ib-9)), 3-(2-pyridyldithio)propanoic acid (compound (Ib-10)), 2,2'-dithiobis(1H-imidazole) (compound (Ib-11)), 2,2'-dithiobis(1-methyl-1H-imidazole) (compound (Ib-12)), 2-phenylisothiazole [5,4-b]pyridine-3(2H)-one (compound (Ic-1)), 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-2)), 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one (compound (Ic-3)), 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-4)), 5-nitro-2-phenyl-1,2-benzothiazole-3(2H)-one (compound (Ic-5)), 2-(2,6-dimethylphenyl It is at least one selected from the group consisting of 5-nitro-1,2-benzothiazole-3(2H)-one (compound (Ic-6)), 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-7)), 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-8)), and 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3(2H)-one (compound (Ic-9)).

[0411] In another embodiment of the manufacturing method of the present invention, compound (I) is preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)), 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)), 2,2'-dibenzimidazolyl disulfide (compound (Ib-4)), 2,2'-dibenzoxazolyl disulfide (compound (Ib-5)), 5,5'-di(1,2, 3-Triazolyl) disulfide (compound (Ib-6)), 5,5'-dithiobis(1-phenyl-1H-tetrazole) (compound (Ib-7)), 2,2'-dithiobis(pyridine-N-oxide) (compound (Ib-8)), 4,4'-dipyridyl disulfide (compound (Ib-9)), 3-(2-pyridyldithio)propanoic acid (compound (Ib-10)), 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)), 2-isopr Ropyruisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-2)), 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one (compound (Ic-3)), 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-4)), 5-nitro-2-phenyl-1,2-benzothiazole-3(2H)-one (compound (Ic-5)), 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazole It is at least one selected from the group consisting of ru-3(2H)-one (compound (Ic-6)), 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-7)), 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-8)), and 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3(2H)-one (compound (Ic-9)).

[0412] In another embodiment of the manufacturing method of the present invention, compound (I) is preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)), 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)), 2,2'-dibenzimidazolyl disulfide (compound (Ib-4)), 2,2'-dibenzoxazolyl disulfide (compound (Ib-5) )), 5,5'-di(1,2,3-triazolyl) disulfide (compound (Ib-6)), 5,5'-dithiobis(1-phenyl-1H-tetrazole) (compound (Ib-7)), 2,2'-dithiobis(pyridine-N-oxide) (compound (Ib-8)), 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)), 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one ( Compound (Ic-2)), 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one (Compound (Ic-3)), 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one (Compound (Ic-4)), 5-nitro-2-phenyl-1,2-benzothiazole-3(2H)-one (Compound (Ic-5)), 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazole-3(2H)-one (Compound At least one selected from the group consisting of (Ic-6), 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-7)), 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-8)), and 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3(2H)-one (compound (Ic-9)).

[0413] In another embodiment of the manufacturing method of the present invention, compound (I) is preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)), 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)), 2,2'-dibenzimidazolyl disulfide (compound (Ib-4)), 2,2'-dibenzoxazolyl disulfide (compound (Ib-5)), 5,5'-di(1,2,3-triazolyl) disulfide (compound (Ib-6)), 5 ,5'-Dithiobis(1-phenyl-1H-tetrazole) (compound (Ib-7)), 2,2'-Dithiobis(pyridine-N-oxide) (compound (Ib-8)), 4,4'-Dipyridyl disulfide (compound (Ib-9)), 3-(2-pyridyldithio)propanoic acid (compound (Ib-10)), 2,2'-Dithiobis(1H-imidazole) (compound (Ib-11)), 2,2'-Dithiobis(1-methyl-1H-imidazole) (compound (Ib-12)), 2-Phenylisothiazolo[5,4-b]pyridine-3(2H)- ONE (compound (Ic-1)), 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-ONE (compound (Ic-2)), 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-ONE (compound (Ic-3)), 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-ONE (compound (Ic-4)), 5-nitro-2-phenyl-1,2-benzothiazole-3(2H)-ONE (compound (Ic-5)), 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazole-3(2H) -one (compound (Ic-6)), 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-7)), 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one (compound (Ic-8)), 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3(2H)-one (compound (Ic-9)), 2-phenylisothiazolo[4,5-c]pyridine-3(2H)-one (compound (Ic-10)), 2-phenylisothiazolo[5,4-b]Quinoline-3(2H)-one (compound (Ic-11)), 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-12)), 2-phenyl-2H-1,2-benzothiadin-3(4H)-one (compound (Ic-13)), 2-(2,6-dimethylphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-14)), It is at least one selected from the group consisting of 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-15)), 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-16)), and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-17)).

[0414] In another embodiment of the production method of the present invention, compound (I) is preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)), 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)), 2,2'-dibenzimidazolyl disulfide (compound (Ib-4)), 2,2'-dibenzoxazolyl disulfide (compound (Ib-5)), 5,5'-dithiobis(1-phenyl-1H-tetrazo (Compound (Ib-7)), 2,2'-Dithiobis(pyridine-N-oxide) (Compound (Ib-8)), 4,4'-Dipyridyl disulfide (Compound (Ib-9)), 3-(2-Pyridyldithio)propanoic acid (Compound (Ib-10)), 2-Phenylisothiazolo[5,4-b]pyridine-3(2H)-one (Compound (Ic-1)), 2-Isopropylisothiazolo[5,4-b]pyridine-3(2H)-one (Compound (Ic-2)), 5-Nitro-2-phenyl-1,2-benzothiazole-3 (2H)-one (compound (Ic-5)), 2-phenylisothiazolo[4,5-c]pyridine-3(2H)-one (compound (Ic-10)), 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one (compound (Ic-11)), 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-12)), 2-phenyl-2H-1,2-benzothiadin-3(4H)-one (compound (Ic-13)), 2-(2,6-dimethylphenyl) It is at least one selected from the group consisting of isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-14)), 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-15)), 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-16)), and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-17)).

[0415] In another embodiment of the production method of the present invention, compound (I) is preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)), 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-2)), 2-phenylisothiazolo[4,5-c]pyridine-3(2H)-one (compound (Ic-10)), 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one (compound (Ic-11)), 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one (compound At least one selected from the group consisting of (Ic-12), 2-phenyl-2H-1,2-benzothiadin-3(4H)-one (compound (Ic-13)), 2-(2,6-dimethylphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-14)), 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-15)), 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-16)), and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-17)).

[0416] In another embodiment of the production method of the present invention, compound (I) is preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)), 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)), 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-12)), 2-phenyl-2H-1,2-benzothiadin-3(4H)-one (compound (Ic-13)), 2-(2,6-dimethylphenyl) It is at least one selected from the group consisting of isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-14)), 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-15)), 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-16)), and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-17)).

[0417] In the production method of the present invention, compound (I) is particularly preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)) and / or 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)), and most preferably 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-1)).

[0418] 2-1-4. Method for producing compound (I) Compound (I) may be a commercially available product or may be prepared by known methods. For example, compound (Ib-1) (i.e., 2,2'-dibenzothiazolyl disulfide, MBTS) is commercially available.

[0419] A typical method for producing compound (I) is shown below, but the method is not limited to these. Compound (I) can be produced by the following representative production methods, the synthesis examples described later, known methods (for example, the method described in the Supporting Information of Non-Patent Document 2), or methods similar thereto. The following are schematic diagrams of the reactions in each step of a typical manufacturing method, where each symbol in the diagram has the same meaning as described above.

[0420] (Method for producing compound (Ib)) Compound (Ib) can be produced, for example, by the oxidation and coupling of thiols, as shown in the following formula.

[0421] [ka]

[0422] Examples of oxidizing agents for oxidizing compound (Ib-1s) and compound (Ib-2s) include hydrogen peroxide, iodine, oxygen, N-bromosuccinimide, and N-chlorosuccinimide. One oxidizing agent may be used alone, or two or more may be used in combination.

[0423] The amount of oxidizing agent used is preferably 1 to 10 moles, more preferably 1 to 5 moles, per 1 mole of the total of compound (Ib-1s) and compound (Ib-2s). It is preferable to use compound (Ib-2s) in approximately equal amounts to compound (Ib-1s).

[0424] This reaction is carried out in a suitable solvent that does not inhibit the reaction. Examples of such solvents include ester solvents such as ethyl acetate and isopropyl acetate; polar ether solvents such as 1,4-dioxane and tetrahydrofuran; halogen solvents such as dichloromethane and chloroform; and aromatic solvents such as benzene, toluene, xylene, and mesitylene. The amount of solvent used is preferably 1 to 20 mL, more preferably 3 to 10 mL, per 1 mmol total of compound (Ib-1s) and compound (Ib-2s).

[0425] The reaction temperature is preferably -10°C to 50°C, more preferably 0°C to 40°C, and the reaction time is preferably 1 to 24 hours, more preferably 1 to 10 hours.

[0426] After the reaction, compound (Ib) can be recovered by known methods (e.g., extraction). The recovered compound (Ib) can be purified by known means (e.g., recrystallization, chromatography).

[0427] (Y 1c Method for producing a compound (Ic) in which the compound is a carbonyl group. Y 1c Compound (Ic) in which is a carbonyl group can be produced, for example, by the following process.

[0428] [ka]

[0429] Process A1 This process is for synthesizing the acid chloride of compound (Ic-1s). Specifically, compound (Ic-1s) and an excess amount of thionyl chloride (for example, 10 moles per mole of compound (Ic-1s)) are added to a reactor, and the mixture is refluxed for 1 hour to 1 night to obtain the acid chloride of compound (Ic-1s).

[0430] This process is preferably carried out under an inert atmosphere (for example, an argon atmosphere). After the reaction is complete, the solid containing the acid chloride of compound (Ic-1s) can be recovered by distilling off the thionyl chloride.

[0431] Process A2 This process involves the acid chloride of compound (Ic-1s) and a suitable amine (R 4c This is a step to synthesize compound (Ic-2s) by reacting it with (-NH2). Specifically, the solid containing the acid chloride of compound (Ic-1s) obtained in step A1, and an appropriate amine (R 4cCompound (Ic-2s) is obtained by adding (-NH2) and a base to a suitable solvent and reacting them.

[0432] The base may be either an organic or inorganic base, and is preferably triethylamine. 4c The amounts of -NH2 and base used are preferably about 1 to 1.5 moles each per mole of the acid chloride of compound (Ic-1s).

[0433] This step is carried out in a suitable solvent that does not inhibit the reaction. Examples of such solvents include polar ether solvents such as 1,4-dioxane and tetrahydrofuran (THF), with THF being preferred. The amount of solvent used is not particularly limited as long as it can dissolve the solid, but is preferably about 5 to 10 mL per 1 mmol of the acid chloride of compound (Ic-1s).

[0434] This process is preferably carried out at a low temperature (e.g., around 0°C). The completion of the reaction can be confirmed by TLC or the like. After the reaction is complete, the compound (Ic-2s) can be recovered by known means (e.g., concentration and extraction).

[0435] Process A3 This process involves the synthesis of compound (Ic-3s) by reacting compound (Ic-2s) with tert-butylthiol. More specifically, compound (Ic-3s) is obtained by reacting compound (Ic-2s) and tert-butylthiol in a suitable solvent in the presence of a base.

[0436] The amount of tert-butylthiol used is preferably about 1 to 1.5 moles per mole of compound (Ic-2s).

[0437] Examples of bases used in this process include alkali metal hydroxides such as sodium hydroxide and potassium hydroxide, with sodium hydroxide being preferred. The amount of base used is preferably about 1 to 2 moles per mole of compound (Ic-3s).

[0438] This step is carried out in a suitable solvent that does not inhibit the reaction. Examples of such solvents include amide solvents such as dimethylformamide (DMF), dimethylacetamide, and N-methylpiperidone, with DMF being preferred. The amount of solvent used is not particularly limited as long as it can dissolve the compound (Ic-2s), but is preferably about 5 to 10 mL per 1 mmol of compound (Ic-2s).

[0439] The reaction temperature in this process is approximately room temperature (e.g., 25°C), and the reaction time is, for example, 1 hour to 1 night, preferably 3 to 20 hours. After the reaction is complete, the compound (Ic-3s) can be recovered by known means (e.g., solid-liquid separation).

[0440] Process A4 This process involves a ring-closing reaction of the compound (Ic-3s), resulting in Y 1c This is a step to synthesize compound (Ic) in which is a carbonyl group. This step can be carried out in two ways, as described in steps A4-1 and A4-2 below.

[0441] Process A4-1 In this process, compound (Ic-3s), dimethyl sulfoxide (DMSO), and chlorotrimethylsilane (TMSCl) are added to a suitable solvent, and the resulting reaction mixture is reacted at room temperature (e.g., 25°C), thereby producing Y 1c We synthesize a compound (Ic) in which is a carbonyl group.

[0442] The amount of DMSO used is preferably about 1 to 2 moles per mole of compound (Ic-3s). The amount of TMSCl used is preferably about 1 to 1.5 moles per mole of compound (Ic-3s).

[0443] This step is carried out in a suitable solvent that does not inhibit the reaction. Examples of the solvent include halogenated solvents such as chloroform, dichloromethane, 1,2-dichloroethane, etc., and dichloromethane is preferred. The amount of the solvent used is not particularly limited as long as compound (Ic-3s) can be dissolved, but it is preferably about 2 - 5 mL per 1 mmol of compound (Ic-3s).

[0444] The reaction time for this step is, for example, 1 hour to overnight, preferably about 5 - 10 hours. After the completion of the reaction, compound (Ic) in which Y 1c is a carbonyl can be recovered by known means (such as solid-liquid separation). The recovered compound (Ic) can be purified by known means (such as recrystallization, chromatography, etc.).

[0445] Step A4-2 In this step, compound (Ic-3s) and meta-chloroperbenzoic acid (mCPBA) are reacted in a suitable solvent to obtain a sulfoxide intermediate, and then by heating this, compound (Ic) in which Y 1c is a carbonyl is synthesized.

[0446] The amount of mCPBA used is preferably about 1 - 1.5 mol per 1 mol of compound (Ic-3s).

[0447] The reaction between compound (Ic-3s) and meta-chloroperbenzoic acid (mCPBA) is carried out in a suitable solvent that does not inhibit the reaction. Examples of the solvent include halogenated solvents such as chloroform, dichloromethane, 1,2-dichloroethane, etc., and dichloromethane is preferred. The amount of the solvent used is not particularly limited as long as compound (Ic-3s) can be dissolved, but it is preferably about 8 - 15 mL per 1 mmol of compound (Ic-3s).

[0448] The reaction between the compound (Ic-3s) and metachloroperbenzoic acid (mCPBA) is preferably carried out at a low temperature (e.g., around 0°C). The reaction time is, for example, about 30 minutes to 1 hour. After the reaction is complete, the sulfoxide intermediate can be recovered by known means (e.g., extraction followed by concentration and recrystallization).

[0449] The recovered sulfoxide intermediate is heated in a suitable solvent that does not inhibit its cyclization reaction, Y 1c Compound (Ic) in which is a carbonyl compound can be synthesized. Examples of the solvent include toluene, xylene, pyridine, and combinations thereof, with a combination of toluene and pyridine being preferred. The reaction temperature is preferably 70°C to 120°C. The synthesized compound (Ic) can be purified by known means (e.g., recrystallization, chromatography, etc.).

[0450] The aforementioned compound (Ic-2s) (i.e., Y 1c An intermediate for synthesizing a compound (Ic) in which is a carbonyl group can also be produced, for example, by the following steps.

[0451] [ka]

[0452] Process A2' This process uses a condensing agent to combine the compound (Ic-1s) with a suitable amine (R 4c This is a step in which the compound (Ic-2s) is synthesized by reacting it with (NH2).

[0453] Examples of condensing agents used in this process include dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC) and its hydrochloride salt (EDC·HCl), hexafluorophosphate (benzotriazole-1-yloxy)tripyrrolidinophosphonium (PyBop), O-(benzotriazole-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU), 1-[bis(dimethylamino)methylene]-5-chloro-1H-benzotriazolium-3-oxide hexafluorophosphate (HCTU), and O-benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU). Among these, EDC·HCl is preferred.

[0454] R 4c The amounts of -NH2 and the condensing agent used are preferably about 1 to 5 moles each per mole of compound (Ic-1s).

[0455] A base may be used in this process. The base can be either an organic or inorganic base, and triethylamine is preferred. When a base is used, the amount is preferably about 1 to 5 moles per mole of compound (Ic-1s).

[0456] This step is carried out in a suitable solvent that does not inhibit the reaction. Examples of such solvents include polar ether solvents such as 1,4-dioxane and tetrahydrofuran (THF), with THF being preferred. The amount of solvent used is not particularly limited, but is preferably about 1 to 10 mL per 1 mmol of compound (Ic-1s).

[0457] The reaction temperature for this step is approximately room temperature (e.g., 25°C), and the reaction time is, for example, 1 hour to 1 night, preferably 3 to 20 hours. This step is preferably carried out under an inert atmosphere (e.g., an argon atmosphere). The completion of the reaction can be confirmed by TLC or the like. After the reaction is complete, the compound (Ic-2s) can be recovered by known means (e.g., concentration and extraction).

[0458] 2-2. Oxidation The present invention is characterized by oxidizing an oligonucleotide precursor having a phosphite ester bond or a phosphonic acid ester bond (hereinafter sometimes abbreviated as "oligonucleotide precursor") using compound (I) (excluding 2,2-dipyridyl disulfide).

[0459] By using compound (I) as an oxidizing agent, the formation of by-products (defective products and desulfurized products) can be suppressed. In order to suppress the formation of by-products, the amount of compound (I) relative to the total amount of oxidizing agent used in the production method of the present invention is preferably 80 mol% or more, more preferably 90 mol% or more. In the production method of the present invention, the oxidizing agent is more preferably compound (I).

[0460] From the viewpoint of ensuring the completion of the oxidation reaction, the amount of compound (I) used is preferably 1 to 15 mol, more preferably 1 to 10 mol, and even more preferably 1 to 5 mol per mol of oligonucleotide precursor.

[0461] The oligonucleotide precursor is preferably, (1) Oligonucleotide precursors having phosphite bonds, obtained by condensation of a nucleoside, nucleotide, or oligonucleotide with a phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide, or (2) Oligonucleotide precursors having phosphite bonds obtained by condensation of oligonucleotides with phosphoramidites Examples of the oligonucleotide precursors in (1) include compound (cI) (i.e., an oligonucleotide precursor having a phosphite ester bond obtained by the condensation of compound (aI) and compound (bI)) and compound (c-I') (i.e., an oligonucleotide precursor having a phosphite ester bond obtained by the condensation of compound (a-I') and compound (b-I')). Examples of the oligonucleotide precursors in (2) include compound (γ-I) and compound (γ-I').

[0462] Oligonucleotide precursors having phosphite ester bonds are more preferably oligonucleotide precursors having phosphite ester bonds obtained by condensation of a nucleoside, nucleotide, or oligonucleotide with a phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide.

[0463] The oligonucleotide precursor is more preferably an oligonucleotide precursor having a phosphite bond, obtained by condensation of a nucleoside, nucleotide, or oligonucleotide (a) having a hydrophobic protecting group with a nucleoside, nucleotide, or oligonucleotide (b) whose hydroxyl group is protected by a temporary protecting group that can be removed under acidic conditions and is phosphoramidized.

[0464] The oligonucleotide precursor is particularly preferably compound (cI) in which Pg is **LYZ (i.e., an oligonucleotide precursor having a phosphite ester bond obtained by the condensation of compound (aI) in which Pg is **LYZ and compound (bI)), or compound (c-I') in which Pg is **LYZ (i.e., an oligonucleotide precursor having a phosphite ester bond obtained by the condensation of compound (a-I') in which Pg is **LYZ and compound (b-I')).

[0465] The oligonucleotide precursor is preferably an oligonucleotide precursor having a thiophosphate ester bond in addition to a phosphite ester bond or a phosphonic acid ester bond. When an oligonucleotide precursor having a thiophosphate ester bond is oxidized with iodine, a by-product (desulfurized product) is produced in which the thiophosphate ester bond is converted to a phosphate ester bond. However, according to the present invention, which uses compound (I) as an oxidizing agent, the production of this desulfurized product can be suppressed.

[0466] It is preferable to oxidize the oligonucleotide precursor in the presence of water. The water present in the system acts as a source of oxygen atoms, thus promoting the oxidation of the oligonucleotide precursor. The amount of water is preferably 0.5 to 200 moles, more preferably 1 to 100 moles, and even more preferably 1 to 50 moles per mole of oligonucleotide precursor.

[0467] The reaction temperature for the oxidation of the oligonucleotide precursor is preferably 10 to 50°C, more preferably 15 to 40°C, and the reaction time is preferably 5 minutes to 5 hours, more preferably 30 minutes to 3 hours.

[0468] The oxidation of oligonucleotide precursors is preferably carried out in a solution containing a nonpolar solvent. The concentration of oligonucleotide precursors in the solution is not particularly limited as long as they are dissolved in the solvent, but is preferably 1 to 30% by weight.

[0469] Examples of nonpolar solvents include halogenated solvents such as chloroform, dichloromethane, and 1,2-dichloroethane; 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 nonpolar ether solvents such as diethyl ether, cyclopentyl methyl ether, and tert-butyl methyl ether. A single nonpolar solvent may be used, or two or more may be used in combination. The nonpolar solvent is preferably at least one selected from the group consisting of halogenated solvents, aromatic solvents, ester solvents, and aliphatic solvents, more preferably halogenated solvents and toluene, even more preferably at least one selected from the group consisting of chloroform, dichloromethane, and toluene, and particularly preferably dichloromethane.

[0470] The solution containing the nonpolar solvent may further contain an aprotic polar solvent. Examples of aprotic polar solvents include nitrile solvents such as acetonitrile and propionitrile; polar ether solvents such as tetrahydrofuran; and pyridine. Among these, nitrile solvents are preferred, and acetonitrile or pyridine are more preferred. The amount of aprotic polar solvent used is preferably 10 to 100 mL, more preferably 10 to 50 mL, per 100 mL of the nonpolar solvent.

[0471] 2-3. One-pot synthesis The manufacturing method of the present invention can also be carried out by one-pot synthesis. This one-pot synthesis is Step (1) involves condensing a nucleoside, nucleotide, or oligonucleotide (a) having a hydrophobic protecting group with a nucleoside, nucleotide, or oligonucleotide (b) that is protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions and is phosphoramidized, in a solution containing a nonpolar solvent, to form an oligonucleotide precursor (c) having a phosphite ester bond and a hydrophobic protecting group, and protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions. Step (2) involves adding a quenching agent (i) for the phosphoramidized nucleoside, nucleotide, or oligonucleotide (b) to the solution after step (1) to quench the phosphoramidized nucleoside, nucleotide, or oligonucleotide (b). Step (3) involves adding an oxidizing agent to the solution after step (2) to oxidize the oligonucleotide precursor (c) to form an oligonucleotide (d) having a phosphate ester bond and a hydrophobic protecting group, and being protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions. Step (4) involves adding an oxidizing agent quenching agent (ii) to the solution after step (3) to quench the oxidizing agent. Step (5) involves adding an acid to the solution after step (4) to remove the temporary protecting group of the oligonucleotide (d) that can be removed under acidic conditions, thereby forming an oligonucleotide (e) having an unprotected hydroxyl group and a hydrophobic protecting group. If necessary, step (6) to add a base to the solution after step (5), and (7) A step in which a polar solvent is added to the solution containing the oligonucleotide (e) to precipitate the oligonucleotide (e). Including, and The oxidizing agent is characterized by being compound (I) (excluding 2,2-dipyridyl disulfide). Herein, the method of production of the present invention by one-pot synthesis is characterized by not isolating the intermediates (i.e., the oligonucleotide precursor (c) obtained in step (1), and the oligonucleotide (d) obtained in step (3)). The steps of the one-pot synthesis will be described in order below.

[0472] (Step (1) (condensation)) In step (1), a nucleoside, nucleotide, or oligonucleotide (a) having a hydrophobic protecting group is condensed with a nucleoside, nucleotide, or oligonucleotide (b) whose hydroxyl group is protected by a temporary protecting group that can be removed under acidic conditions and is phosphoramidized in a solution containing a nonpolar solvent to form an oligonucleotide precursor (c) having a phosphite ester bond and a hydrophobic protecting group, and whose hydroxyl group is protected by a temporary protecting group that can be removed under acidic conditions.

[0473] The hydrophobic protecting group possessed by the nucleoside, nucleotide, or oligonucleotide (a) is the protecting group (Pg-5). The description of the protecting group (Pg-5) is as described above.

[0474] The primary protecting group of the hydroxyl group of the phosphoramidized nucleoside, nucleotide, or oligonucleotide (b) is preferably a 4,4'-dimethoxytrityl group or a 4-monomethoxytrityl group, and more preferably a 4,4'-dimethoxytrityl group, from the viewpoint of ease of deprotection and other factors.

[0475] The combination of the nucleoside, nucleotide, or oligonucleotide (a) used in step (1) and the phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide (b) is preferably a combination of compound (aI) and compound (bI), or a combination of compound (a-I') and compound (b-I'), more preferably a combination of compound (aI) in which Pg is **LYZ and compound (bI), or a combination of compound (a-I') in which Pg is **LYZ and compound (b-I').

[0476] Step (1) is carried out in a solution containing a nonpolar solvent. Examples of nonpolar solvents include halogenated solvents such as chloroform, dichloromethane, and 1,2-dichloroethane; 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 nonpolar ether solvents such as diethyl ether, cyclopentyl methyl ether, and tert-butyl methyl ether. Only one nonpolar solvent may be used, or two or more may be used in combination. Preferably, the nonpolar solvent is at least one selected from the group consisting of halogenated solvents, aromatic solvents, ester solvents, and aliphatic solvents, more preferably at least one selected from the group consisting of halogenated solvents and toluene, even more preferably at least one selected from the group consisting of chloroform, dichloromethane, and toluene, and particularly preferably dichloromethane and / or toluene. The steps after step (1) are also carried out in a solution containing a nonpolar solvent.

[0477] The solution containing the nonpolar solvent may further contain an aprotic polar solvent. Examples of aprotic polar solvents include nitrile solvents such as acetonitrile and propionitrile; polar ether solvents such as tetrahydrofuran; among these, nitrile solvents are preferred, and acetonitrile is more preferred. The amount of aprotic polar solvent used is preferably 10 to 100 mL, more preferably 10 to 50 mL, per 100 mL of the nonpolar solvent.

[0478] The amount of phosphoramidized nucleoside, nucleotide, or oligonucleotide (b) used is, for example, 1 to 10 mol, preferably 1 to 5 mol, per 1 mol of nucleoside, nucleotide, or oligonucleotide (a) used. The concentration of the nucleoside, nucleotide, or oligonucleotide (a) in the solution is preferably 1 to 30% by weight.

[0479] The reaction temperature in step (1) is not particularly limited as long as condensation proceeds, but is for example 0 to 100°C, preferably 20 to 50°C. The reaction time varies depending on the type of raw materials used, the reaction temperature, etc., but is for example 5 minutes to 24 hours.

[0480] To promote condensation, known activators may be used. Examples of activators include pyridine trifluoroacetate, tetrazole, 5-ethylthio-1H-tetrazole, 5-benzylthio-1H-tetrazole, and 4,5-dicyanoimidazole. Only one activator may be used, or two or more may be used in combination. When an activator is used, the amount used is preferably 0.5 to 10 mol, more preferably 1 to 5 mol, per 1 mol of the phosphoramidized nucleoside, nucleotide, or oligonucleotide (b).

[0481] (Step (2) (Quenching of phosphoramidized nucleoside, nucleotide, or oligonucleotide (b))) In step (2), the quenching agent (i) for the phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide (b) is added to the solution after step (1) to quench the phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide (b).

[0482] Quenching agent (i) may be used alone or in combination of two or more. Examples of quenching agent (i) include water, alcohols, phenols, and amines.

[0483] Examples of alcohols that can be used as a quenching agent (i) include halogenated monohydric alcohols such as methanol, 2-propanol, t-butanol, 2,2,2-trifluoroethanol, tetrahydrofurfuryl alcohol, furfuryl alcohol, 2,3-O-isopropylidene-D-ribofuranose, and 3'-O-triisopropylsilyl-thymidine, and halogenated polyhydric alcohols such as ethylene glycol and diethylene glycol.

[0484] Examples of phenols that can be used as quenching agents (i) include 4-nitrophenol and pentafluorophenol. Examples of amines that can be used as quenching agents (i) include morpholine.

[0485] The amount of quenching agent (i) used is preferably 1 to 20 mol, more preferably 1 to 10 mol, and even more preferably 1 to 5 mol, relative to 1 mol of phosphoramidized nucleoside, nucleotide, or oligonucleotide (b) used in step (1).

[0486] The quenching agent (i) is preferably water. Using water as the quenching agent (i) can accelerate the oxidation in the subsequent step (3). When water is used as the quenching agent (i), the amount used is preferably 1 to 20 mol, more preferably 2 to 15 mol, and even more preferably 2 to 10 mol, per 1 mol of phosphoramidized nucleoside, nucleotide, or oligonucleotide (b) used in step (1).

[0487] The temperature of the solution after adding the quenching agent (i) is not particularly limited as long as the phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide (b) can be quenched, but is preferably 5 to 40°C, more preferably 15 to 30°C. The stirring time of the solution after adding the quenching agent (i) varies depending on the type of quenching agent (i) used, the temperature, etc., but is for example 10 minutes to 3 hours.

[0488] (Step (3) (oxidation)) Step (3) is characterized by using compound (I) (excluding 2,2-dipyridyl disulfide) as an oxidizing agent to oxidize the phosphite ester bond of the oligonucleotide precursor (c) and convert it to a phosphate ester bond. Specifically, in step (3), an oxidizing agent (i.e., compound (I)) is added to the solution after step (2) to oxidize the oligonucleotide precursor (c) to form an oligonucleotide (d) having a phosphate ester bond and a hydrophobic protecting group, and protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions. The explanation of the oxidizing agent and oxidation is as described above.

[0489] In the manufacturing method of the present invention, the formation of desulfurized products can be suppressed, therefore, the oligonucleotide precursor (c) is preferably an oligonucleotide precursor having a thiophosphate ester bond in addition to a phosphite ester bond.

[0490] (Addition of aromatic amines) It is preferable to add an aromatic amine to the solution after step (3). Only one aromatic amine may be used, or two or more may be used in combination. Adding an aromatic amine can further suppress the formation of defective bodies. The aromatic amine may be added after step (3) and before the addition of the quenching agent in step (4), or the quenching agent (ii) and the aromatic amine may be added simultaneously in step (4), or the quenching agent (ii) and the aromatic amine may be added sequentially in step (4).

[0491] Examples of aromatic amines include aniline, 2-chloroaniline, 3-chloroaniline, 2,4-dichloroaniline, 2-fluoroaniline, 4-methoxyaniline, 4-nitroaniline, 2,6-dichloroaniline, and 2,6-xylidine.

[0492] The aromatic amine is preferably at least one selected from the group consisting of aniline, 2-chloroaniline, 3-chloroaniline, 2,4-dichloroaniline, 2-fluoroaniline, 4-methoxyaniline, 4-nitroaniline, 2,6-dichloroaniline, and 2,6-xylidine, more preferably 2-chloroaniline and / or 2,6-xylidine, and even more preferably 2,6-xylidine (i.e., 2,6-dimethylaniline).

[0493] From the viewpoint of suppressing defects, the amount of aromatic amine used is preferably 1 to 20 mol, more preferably 1 to 10 mol, and even more preferably 1 to 5 mol, per 1 mol of oxidizing agent used in step (3).

[0494] (Step (4) (Quenching of the oxidizing agent)) In step (4), the oxidizing agent quenching agent (ii) is added to the solution after step (3) to quench the oxidizing agent. Quenching agent (ii) may be used alone or in combination of two or more types.

[0495] The quenching agent (ii) is preferably an organophosphorus compound. One organophosphorus compound may be used alone, or two or more may be used in combination.

[0496] The organophosphorus compound is preferably at least one selected from the group consisting of phosphines, phosphite triesters, phosphiniate esters, phosphonic acid diesters, and phosphiniate esters, and more preferably phosphines. The phosphines, etc., may be used individually or in combination of two or more.

[0497] Phosphines are compounds represented by the formula P(R)3 (wherein the formula, the three Rs each independently represent a hydrogen atom, an alkyl group, or an aryl group). Examples of phosphines include triphenylphosphine and methyldiphenylphosphine.

[0498] Triethyl phosphite is a compound represented by the formula P(OR')3 (wherein the three R' groups each independently represent an alkyl group or an aryl group). Examples of triethyl phosphite include triethyl phosphite.

[0499] Phosphosphinate esters are compounds represented by the formula: P(OR')(R)2 (wherein the formula, the two Rs each independently represent a hydrogen atom, an alkyl group, or an aryl group, and R' represents an alkyl group or an aryl group). Examples of phosphosphinate esters include ethoxydiphenylphosphine.

[0500] Phosphosphite diesters are given by formula: P(R)(OR') 2、 This is a compound represented by the formula (wherein R represents a hydrogen atom, an alkyl group, or an aryl group, and the two R's each independently represent an alkyl group or an aryl group). Examples of phosphonites include diethoxyphenylphosphine.

[0501] Phosphinic acid esters are compounds represented by the formula: P(=O)(R)2(OR') (wherein the formula, the two Rs each independently represent a hydrogen atom, an alkyl group, or an aryl group, R' represents an alkyl group or an aryl group, and one R and R' may form a heterocycle together with the phosphorus and oxygen atoms to which they are bonded). Examples of phosphinic acid esters include 9,10-dihydro-9-oxa-10-phosphaphenanthrene 10-oxide, represented by the following formula.

[0502] [ka]

[0503] In one embodiment of the manufacturing method of the present invention, the quenching agent (ii) is preferably at least one selected from the group consisting of triphenylphosphine, methyldiphenylphosphine, triethyl phosphite, ethoxydiphenylphosphine, diethoxyphenylphosphine, and 9,10-dihydro-9-oxa-10-phosphaphenanthrene 10-oxide, more preferably triphenylphosphine and / or methyldiphenylphosphine, and even more preferably triphenylphosphine.

[0504] The amount of quenching agent (ii) used should be sufficient to quench the excess oxidizing agent in step (3), and is preferably 1 to 10 mol, more preferably 1 to 5 mol, per 1 mol of the excess oxidizing agent (i.e., "amount of oxidizing agent used in step (3)" - "amount of phosphoramiditized nucleoside, nucleotide, or oligonucleotide (b) used in step (1)").

[0505] The temperature of the solution after adding the quenching agent (ii) is not particularly limited as long as the oxidizing agent is quenched, but is preferably 0°C to 50°C, more preferably 10°C to 40°C. The stirring time of the solution after adding the quenching agent (ii) varies depending on the type of quenching agent (ii) used, temperature, etc., but is preferably 5 minutes to 5 hours, more preferably 5 minutes to 2 hours.

[0506] (Step (5) (Removal of temporary protecting group)) In step (5), an acid is added to the solution after step (4) to remove the temporary protecting group of oligonucleotide (d) that can be removed under acidic conditions, thereby forming oligonucleotide (e) having an unprotected hydroxyl group and a hydrophobic protecting group. One acid may be used alone, or two or more acids may be used in combination.

[0507] The acid is not particularly limited as long as it can effectively remove the temporary protecting group, but examples include trifluoroacetic acid, dichloroacetic acid, trifluoromethanesulfonic acid, trichloroacetic acid, methanesulfonic acid, hydrochloric acid, acetic acid, and p-toluenesulfonic acid. From the viewpoint of effectively removing the temporary protecting group, trifluoroacetic acid, dichloroacetic acid, trifluoromethanesulfonic acid, and trichloroacetic acid are more preferred, trifluoroacetic acid, dichloroacetic acid, and trifluoromethanesulfonic acid are even more preferred, trifluoroacetic acid and trifluoromethanesulfonic acid are even more preferred, and trifluoroacetic acid is particularly preferred.

[0508] The amount of acid used is, for example, 1 to 100 moles, preferably 1 to 40 moles, per mole of oligonucleotide(d).

[0509] The reaction temperature in step (5) is not particularly limited as long as the reaction proceeds, but is for example -10°C to 50°C, more preferably 0°C to 40°C. The reaction time varies depending on the oligonucleotide (d) used, the type of acid and the type of nonpolar solvent, the reaction temperature, etc., but is for example 5 minutes to 5 hours.

[0510] It is preferable to add the cation scavenger to the solution before, during, or after the removal of the temporary protecting group of oligonucleotide (d). That is, it is preferable to remove the temporary protecting group in the presence of the cation scavenger, or to add the cation scavenger to the reaction solution after the removal of the temporary protecting group. Only one type of cation scavenger may be used, or two or more types may be used in combination.

[0511] The cation scavenger is not particularly limited as long as it does not cause reprotection by the removed temporary protecting group or a side reaction to the deprotected functional group, but any pyrrole derivatives such as pyrrole, 2-methylpyrrole, 3-methylpyrrole, 2,3-dimethylpyrrole, and 2,4-dimethylpyrrole; indole derivatives such as indole, 3-methylindole, 4-methylindole, 5-methylindole, 6-methylindole, 7-methylindole, 5,6-dimethylindole, 6,7-dimethylindole, and 5-methoxyindole; and furan derivatives such as 2-methylfuran, 2,3-dimethylfuran, 2-methyl-3-(methylthio)furan, and mentfuran can be used. The amount of cation scavenger used is preferably 1 to 50 mol, more preferably 5 to 20 mol, per 1 mol of oligonucleotide(d).

[0512] (Step (6) (neutralization)) To neutralize the acid used in step (5), the one-pot synthesis may further include step (6), in which a base is added to the solution after step (5) and before step (7). However, since the acid used in step (5) can be removed from oligonucleotide(e) by performing step (7) (solid-liquid separation) and washing, step (6) (neutralization) is not essential.

[0513] A single base may be used, or two or more bases may be used in combination. Organic bases are preferred as the bases. Examples of organic bases include pyridine, 2,4,6-trimethylpyridine, benzimidazole, 1,2,4-triazole, N-phenylimidazole, 2-amino-4,6-dimethylpyrimidine, 1,10-phenanthroline, imidazole, N-methylimidazole, 2-chlorobenzimidazole, 2-bromobenzimidazole, 2-methylimidazole, 2-phenylbenzimidazole, N-phenylbenzimidazole, and 5-nitrobenzimidazole. Among these, pyridine, 2,4,6-trimethylpyridine, benzimidazole, 1,2,4-triazole, N-phenylimidazole, N-methylimidazole, 2-amino-4,6-dimethylpyrimidine, and 1,10-phenanthroline are preferred; pyridine, 2,4,6-trimethylpyridine, benzimidazole, 1,2,4-triazole, and N-phenylimidazole are more preferred; pyridine, 2,4,6-trimethylpyridine, benzimidazole, and 1,2,4-triazole are even more preferred; and pyridine, 2,4,6-trimethylpyridine, and benzimidazole are particularly preferred.

[0514] The amount of base used in step (6) is preferably 1 to 10 moles, and more preferably 1 to 3 moles, relative to the amount of acid used in step (5) (1 mole).

[0515] (Step (7) (solid-liquid separation)) In step (7), a polar solvent is added to the solution containing the oligonucleotide (e) (i.e., the solution after step (5), or the solution after step (6) if necessary) to precipitate the oligonucleotide (e).

[0516] Examples of polar solvents used in step (7) include alcohol-based solvents such as methanol, ethanol, and isopropanol; nitrile-based solvents such as acetonitrile and propionitrile; ketone-based solvents such as acetone and 2-butanone; polar ether-based solvents such as 1,4-dioxane and tetrahydrofuran; amide-based solvents such as dimethylformamide, dimethylacetamide, and N-methylpiperidone; sulfoxide-based solvents such as dimethyl sulfoxide; and water. One polar solvent may be used alone, or two or more may be used in combination. Among these, nitrile-based solvents are preferred, and acetonitrile is more preferred.

[0517] To increase the recovery rate of oligonucleotide(e), the amount of polar solvent added in the solid-liquid separation is preferably 1 to 20 mL, more preferably 5 to 20 mL, and even more preferably 5 to 10 mL, per 1 mL of nonpolar solvent contained in the solution.

[0518] To increase the recovery rate of oligonucleotide(e), a precipitation accelerator such as that described in International Publication No. 2016 / 117663 (e.g., 3,4,5-tris(octadecyloxy)benzylpivalate) may be used.

[0519] The precipitated oligonucleotide(e) can be recovered by known means such as filtration.

[0520] The oligonucleotide chain can be extended by repeating a one-pot synthesis including steps (1) to (5) and (7) (preferably steps (1) to (7)). Such a method for producing oligonucleotides by repeating one-pot synthesis is also included in the production method of the present invention.

[0521] (deprotection) The manufacturing method of the present invention may include a step after step (7) (solid-liquid separation) in which protecting groups (e.g., phosphate protecting groups, hydrophobic protecting groups) of the obtained oligonucleotide (e) are removed by known methods. For example, the 2-cyanoethyl group, which is a phosphate protecting group, and the aforementioned protecting groups (Pg-5), etc., can be removed by treatment with aqueous ammonia, aqueous ammonia / ethanol solution, or a mixture of aqueous ammonia and aqueous methylamine solution.

[0522] 3.New compounds The present invention relates to a method for producing oligonucleotides, which is useful in formula (In):

[0523] [ka]

[0524] [In the formula, (1)R 1n represents an optionally substituted phenyl group, p' represents 0, ring C'' represents a 10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom, and q' represents 0 or (2)R 1n represents an optional phenyl group, p' represents 0, ring C'' represents a 6-membered nitrogen-containing aromatic heterocycle, q' represents 1, and R 3n is C 1-6 Does it indicate a perfluoroalkyl group? (3)R 1n represents an optional phenyl group, p' represents 1, ring C'' represents a 6-membered nitrogen-containing aromatic heterocycle, and q' represents 0, or (4)R 1n is one C 1-6 This indicates a pyridyl group substituted with an alkyl group, where p' represents 0, the ring C'' represents a 6-membered nitrogen-containing aromatic heterocycle, and q' represents 0. We also provide compounds represented by [formula].

[0525] Compound (In) is useful as an oxidizing agent for oxidizing oligonucleotide precursors having phosphite ester bonds or phosphonic acid ester bonds (particularly oligonucleotide precursors having phosphite ester bonds) in the production of oligonucleotides. Compound (In) may be used alone or in combination of two or more types.

[0526] In equation (In), "p' represents 0" means that the parenthesized -CH2- in equation (In) does not exist. In equation (In), "q' represents 0" means that R in equation (In) 3n This means that it does not exist.

[0527] R 1n The "optionally substituted phenyl group" is preferably a phenyl group. R 1n "1 C" 1-6 The pyridyl group substituted with an alkyl group is preferably a pyridyl group substituted with one methyl group, and more preferably a 4-methyl-2-pyridyl group.

[0528] The "10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom" of ring C'' is preferably a quinoline ring. The "6-membered nitrogen-containing aromatic heterocycle" of ring C'' is preferably a pyridine ring.

[0529] R 3n C 1-6 The "perfluoroalkyl group" is preferably a trifluoromethyl group.

[0530] Compound (In) is preferably 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one (compound (Ic-11)), 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-12)), 2-phenyl-2H-1,2-benzothiadin-3(4H)-one (compound (Ic-13)), or 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one (compound (Ic-15)).

[0531] The compound (In) is the aforementioned "Y 1c Compounds (Ic) in which the compound is a carbonyl group can be produced by the following methods: synthesis examples described later, known methods (for example, the method described in the Supporting Information of Non-Patent Document 2), or methods similar thereto. [Examples]

[0532] The present invention will be described in more detail below with reference to examples, etc., but the present invention is not limited by the following examples, etc., and can be implemented with appropriate modifications within the scope that is consistent with the spirit of the above and below, and all such modifications are included within the technical scope of the present invention.

[0533] In the following examples, "room temperature" means "20°C to 30°C". Furthermore, the meanings of the abbreviations used in the following examples are as follows.

[0534] DMF: Dimethylformamide THF: Tetrahydrofuran SUC: succinyl TOB: 3,4,5-Tris(octadecyloxy)benzyloxy Piv-TOB:3,4,5-Tris(octadecyloxy)benzylpivalate DDTT:[(N,N-dimethylaminomethylidene)amino]-3H-1,2,4-dithiazoline-3-thion POS:5-phenyl-3H-1,2,4-dithiazol-3-one DMTr:4,4'-Dimethoxytrityl

[0535] (Phosphoramidite or H-phosphonate) 2'-OMe-C-CE phosphoramidite:5'-O-(4,4'-dimethoxytrityl)-N 4 -Benzoyl-2'-O-methylcytidine-3'-[O-(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite dT-CE phosphoramidite: 5'-O-(4,4'-dimethoxytrityl)-deoxythymidine-3'-[O-(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite dC-CE phosphoramidite:5'-O-(4,4'-dimethoxytrityl)-N 4 -Benzoyl-deoxycytidine-3'-[O-(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite 5-Me-dC(Bz)-CE phosphoramidite:5'-O-(4,4'-dimethoxytrityl)-N 4 -Benzoyl-2'-deoxy-5-methylcytidine-3'-[O-(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite dA-CE phosphoramidite:5'-O-(4,4'-dimethoxytrityl)-N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite dG-CE phosphoramidite:5'-O-(4,4'-dimethoxytrityl)-N 2 -Isobutyryl-2'-O-methylguanosine-3'-[O-(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite 2'-FA-CE phosphoramidite:5'-O-(4,4'-dimethoxytrityl)-N 4 -Benzoyl-2'-fluorodeoxyadenosine-3'-[O-(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite dT-H-phosphonate TEA salt: Triethylamine salt of 5'-O-(4,4'-dimethoxytrityl)-deoxythymidine-3'-H-phosphonate

[0536] (oligonucleotide) HO-T-SUC-TOB: Deoxythymidine-3'-yl 3,4,5-tris(octadecyloxy)benzyl succinate HO-T(S)T-SUC-TOB: Deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothinyl Deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-Am(S)Cm-SUC-TOB:N 6 -Benzoyl-2'-O-methyladenosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4 -Benzoyl-2'-O-methylcytidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-Cm(O)Am(S)Cm-SUC-TOB:N 4 -Benzoyl-2'-O-methylcytidine-3'-[O-(2-cyanoethyl)]phosphoryl N 6 -Benzoyl-2'-O-methyladenosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4 -Benzoyl-2'-O-methylcytidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-T(O)T-SUC-TOB: Deoxythymidine-3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-Cm(O)T(O)T-SUC-TOB:N 4 -Benzoyl-2'-O-methylcytidine 3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-Af(O)Cm(O)T(O)T-SUC-TOB:N 6 -Benzoyl-2'-fluorodeoxyadenosine-3'-[O-(2-cyanoethyl)]phosphoryl N 4 -Benzoyl-2'-O-methylcytidine 3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-Af(O)Cm(O)T(O)T-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-N 6 -Benzoyl-2'-fluorodeoxyadenosine-3'-[O-(2-cyanoethyl)]phosphoryl N 4 -Benzoyl-2'-O-methylcytidine 3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-Af(O)Cm(O)T(O)T-OH:5'-O-(4,4'-dimethoxytrityl)-2'-fluorodeoxyadenosine-3'-phosphoryl-2'-O-methylcytidine-3'-phosphoryl-deoxythymidine-3'-phosphoryl-deoxythymidine DMTrO-Af(O)Cm(O)T-OH:5'-O-(4,4'-dimethoxytrityl)-2'-fluorodeoxyadenosine-3'-phosphoryl-2'-O-methylcytidine-3'-phosphoryl-deoxythymidine DMTrO-Af(O)T(O)T-OH:5'-O-(4,4'-dimethoxytrityl)-2'-fluorodeoxyadenosine-3'-phosphoryl-deoxythymidine-3'-phosphoryl-deoxythymidine HO-A(S)C(S)T(S)T-SUC-TOB:N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4-Benzoyl-deoxycytidine 3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-G(III)A(S)C(S)T(S)T-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphytyl N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4 -Benzoyl-deoxycytidine 3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-G(O)A(S)C(S)T(S)T-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphoryl N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4 -Benzoyl-deoxycytidine 3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-G(O)A(S)C(S)T(S)T-OH:5'-O-(4,4'-dimethoxytrityl)-deoxyguanosine-3'-phosphoryl-deoxyadenosine-3'-phospholothionyl-deoxycytidine-3'-phospholothionyl-deoxythymidine-3'-phospholothionyl-deoxythymidine DMTrO-T(OH)T-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-deoxythymidine-3'-phosphoryl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-G(O)T(S)T-SUC-TOB:N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-A(O)G(O)T(S)T-SUC-TOB:N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)]phosphoryl N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO- Me C(O)A(O)G(O)T(S)T-SUC-TOB:N 4 -Benzoyl-2'-deoxy-5-methylcytidine-3'-[O-(2-cyanoethyl)]phosphoryl N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)]phosphoryl N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-T(S)T(S)T-SUC-TOB: Deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl Deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl Deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-T(S)T(S)T-OH:Deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl-deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl-deoxythymidine DMTrO-C(O)T(S)T-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-N 4 -Benzoyl-deoxycytidine 3'-[O-(2-cyanoethyl)]phosphoryl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-C(S)T(S)T-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-N 4 -Benzoyl-deoxycytidine-3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO- Me C-SUC-TOB:N 4 -Benzoyl-5-methyl-2'-deoxycytidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-G(S) Me C-SUC-TOB:N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4 -Benzoyl-5-methyl-2'-deoxycytidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate HO-A(S)G(S) Me C-SUC-TOB:N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4-Benzoyl-5-methyl-2'-deoxycytidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-G(O)A(S)G(S) Me C-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphoryl N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4 -Benzoyl-5-methyl-2'-deoxycytidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-G(S)A(S)G(S) Me C-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4 -Benzoyl-5-methyl-2'-deoxycytidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-G(O)A(S)G(S) Me C-OH:5'-O-(4,4'-dimethoxytrityl)-deoxyguanosine-3'-phosphoryl-deoxyadenosine-3'-phospholothionyl-deoxyguanosine-3'-phospholothionyl-5-methyl-2'-deoxycytidine

[0537] (Alkyl alkyl precursor) DMTrO-G(III)A(S)C(S)T(S)T-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphytyl N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4 -Benzoyl-deoxycytidine 3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphorothionyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-C(III)T(S)T-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-N 4 -Benzoyl-deoxycytidine-3'-[O-(2-cyanoethyl)]phosphytyl deoxythymidine-3'-[O-(2-cyanoethyl)]phosphothionyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-T(H)T-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-deoxythymidine-3'-phosphonyl deoxythymidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMTrO-G(III)A(S)G(S) Me C-SUC-TOB:5'-O-(4,4'-dimethoxytrityl)-N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphytyl N 6 -[1-(dimethylamino)ethylidene]-deoxyadenosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 2 -Isobutyryl-deoxyguanosine-3'-[O-(2-cyanoethyl)]phosphorothionyl N 4 -Benzoyl-5-methyl-2'-deoxycytidine-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate

[0538] Synthesis Example 1: Synthesis of oligonucleotide precursors containing phosphite ester bonds Under an argon atmosphere, HO-T-SUC-TOB (2.5 g, 2.0 mmol), Piv-TOB (5.0 g, 5.0 mmol), anhydrous dichloromethane (100 mL), and anhydrous acetonitrile (30 mL) were added to a 1000 mL flask to prepare a solution. Then, dT-CE phosphoramidite (3.0 g, 4.0 mmol) and triphenylphosphine (270 mg, 1.0 mmol) were added, and the solution was stirred for 30 minutes. Subsequently, 5-ethylthio-1H-tetrazole (530 mg, 4.0 mmol) was added, and the solution was stirred at room temperature for 45 minutes. Then, 2,2,2-trifluoroethanol (1.5 mL, 20 mmol) was added to the solution, and after stirring at room temperature for 30 minutes, 2,6-dimethylaniline (1.8 mL, 15 mmol) and POS (1.0 g, 5.1 mmol) were added sequentially, and the solution was stirred at room temperature for 1.0 hour. Subsequently, 2,3-dimethylfuran (2.1 mL, 20 mmol), trifluoroacetic acid (5.2 mL, 68 mmol), and 2,6-dimethylaniline (83.5 μL, 0.7 μmol) were added sequentially, and the solution was stirred at room temperature for 55 minutes. Further addition of trifluoroacetic acid (620 μL, 8.1 mmol) and 2,6-dimethylaniline (10 μL, 0.10 μmol) was added, and the solution was stirred at room temperature for 15 minutes. Then, pyridine (19 mL, 230 mmol) and methanol (870 μL) were added sequentially, and the solution was stirred at room temperature for 115 minutes. After that, acetonitrile (1 L) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and obtained a mixture of Piv-TOB and the dimer oligonucleotide HO-T(S)T-SUC-TOB (8.1 g, 98% yield). m / z:Calcd. 1610.03,Found 1611.06[M+H] +

[0539] A mixture of Piv-TOB and the trimer oligonucleotide HO-C(S)T(S)T-SUC-TOB was obtained using the same procedure as above, except that the solid obtained above (8.1 g) was used and dC-CE phosphoramidite was used instead of dT-CE phosphoramidite (8.7 g, yield 97%). m / z:Calcd. 2072.11,Found 2073.14[M+H] +

[0540] A mixture of Piv-TOB and the tetramer oligonucleotide HO-A(S)C(S)T(S)T-SUC-TOB was obtained using the same procedure as above, except that the solid obtained above (2.8 g) was used and dA-CE phosphoramidite was used instead of dT-CE phosphoramidite (3.0 g, yield 98%). m / z:Calcd. 2523.23,Found 1262.63[M+2H] 2+

[0541] Under an argon atmosphere, the solid obtained above (400 mg), anhydrous dichloromethane (4.0 mL), and anhydrous acetonitrile (1.2 mL) were added to a 100 mL two-necked flask to prepare a solution. Then, dG-CE phosphoramidite (136 mg, 161 μmol) and 5-ethylthio-1H-tetrazole (21.1 mg, 162 μmol) were added sequentially, and the solution was stirred at room temperature for 60 minutes. After that, acetonitrile (30 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and obtained a mixture of Piv-TOB and the oligonucleotide precursor DMTrO-G(III)A(S)C(S)T(S)T-SUC-TOB, which is a pentamer (453 mg, 99% yield). m / z:Calcd. 3261.49,Found 1631.67[M+2H] 2+

[0542] Example 1: Oxidation using compound (Ib-1) Under an argon atmosphere, the solid obtained in Synthesis Example 1 (100.0 mg: 56.9 mg, 17.4 μmol, as DMTrO-G(III)A(S)C(S)T(S)T-SUC-TOB), anhydrous dichloromethane (0.9 mL), and anhydrous acetonitrile (0.3 mL) were added to a 20 mL two-necked flask to prepare the solution. Then, water (9 μL, 0.5 mmol) and compound (Ib-1) (12.1 mg, 36.4 μmol) were added sequentially, and the solution was stirred at room temperature for 60 minutes. After that, triphenylphosphine (4.6 mg, 18 μmol) was added, and the solution was stirred for 30 minutes. Next, acetonitrile (10 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, then dried to obtain a mixture of Piv-TOB and the pentamer oligonucleotide DMTrO-G(O)A(S)C(S)T(S)T-SUC-TOB (82 mg, yield 82%). Note that a commercially available compound (Ib-1) was used. m / z:Calcd. 3277.48,Found 1639.75[M+2H] 2+

[0543] Example 2: Oxidation using compound (Ic-1) Under an argon atmosphere, the solid obtained in Synthesis Example 1 (100 mg: 57 mg, 17 μmol, as DMTrO-G(III)A(S)C(S)T(S)T-SUC-TOB), anhydrous dichloromethane (0.9 mL), and anhydrous acetonitrile (0.3 mL) were added to a 20 mL two-necked flask to prepare the solution. Then, water (9 μL, 0.5 mmol) and compound (Ic-1) (8.0 mg, 35 μmol) were added sequentially, and the solution was stirred at room temperature for 60 minutes. After that, triphenylphosphine (4.7 mg, 18 μmol) was added, and the solution was stirred for 30 minutes. Next, acetonitrile (10 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel. After drying, a mixture of Piv-TOB and the pentamer oligonucleotide DMTrO-G(O)A(S)C(S)T(S)T-SUC-TOB was obtained (83 mg, yield 83%). The compound (Ic-1) used was synthesized by the method described in the Supporting Information of Non-Patent Document 2. m / z:Calcd.3277.48,Found 1639.75[M+2H] 2+

[0544] Comparative Example 1: Oxidation using iodine Under an argon atmosphere, the solid obtained in Synthesis Example 1 (100 mg: 57 mg as DMTrO-G(III)A(S)C(S)T(S)T-SUC-TOB, 17 μmol), anhydrous dichloromethane (0.9 mL), and anhydrous acetonitrile (0.3 mL) were added to a 20 mL two-necked flask to prepare the solution. Then, water (9 μL, 0.5 mmol), pyridine (5.7 μL, 71 μmol), and iodine (8.8 mg, 35 μmol) were added sequentially, and the solution was stirred at room temperature for 60 minutes. After that, triphenylphosphine (4.8 mg, 18 μmol) was added, and the solution was stirred for 30 minutes. Next, acetonitrile (10 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, then dried to obtain a mixture of Piv-TOB and the pentamer oligonucleotide DMTrO-G(O)A(S)C(S)T(S)T-SUC-TOB (83 mg, yield 83%). m / z:Calcd. 3277.48,Found 1639.75[M+2H] 2+

[0545] Comparison of Example 1, Example 2, and Comparative Example 1 The oligonucleotides containing thiophosphate ester bonds and the by-products obtained when the thiophosphate ester bonds are converted to phosphate ester bonds (hereinafter referred to as "desulfurized products") were analyzed as follows, and the proportion of desulfurized products was calculated.

[0546] The solid (10 mg) obtained in Example 1, Example 2, or Comparative Example 1 and 28% by weight aqueous ammonia (5 mL) were placed in an autoclave and heated at 65°C for 4 hours, then cooled to room temperature. After removing insoluble matter from the reaction mixture using a syringe filter, the mixture was concentrated under reduced pressure using a centrifugal evaporator to obtain the oligonucleotide DMTrO-G(O)A(S)C(S)T(S)T-OH (hereinafter referred to as "the target oligonucleotide") having a thiophosphate ester bond. m / z:Calcd. 1827.36,Found 1826.34[MH] -

[0547] The mixture containing the obtained target oligonucleotide and desulfurized product was analyzed by liquid chromatography-mass spectrometry (LC-MS). Using the extracted ion chromatogram (EIC) of each observed compound (target oligonucleotide and desulfurized product), the peak area of ​​each compound was calculated using the following formula: Percentage of desulfurized product (%) = (Peak area of ​​desulfurized product / Peak area of ​​target oligonucleotide) × 100 The percentage of desulfurized material was calculated using the following method. The results are shown in Table 1.

[0548] [Table 1]

[0549] As is clear from the results in Table 1, the formation of desulfurized products can be suppressed by using compound (Ib-1) or compound (Ic-1) instead of iodine as the oxidizing agent.

[0550] Synthesis Example 2: Synthesis of oligonucleotide precursors containing phosphonic acid ester bonds Under an argon atmosphere, HO-T-SUC-TOB (500 mg, 400 μmol), dT-H-phosphonate TEA salt (568 mg, 800 μmol), anhydrous dichloromethane (10 mL), and anhydrous pyridine (10 mL) were added to a 200 mL three-necked flask to prepare a solution. Subsequently, pivaloyl chloride (0.2 mL, 1.6 mmol) was added, and the solution was stirred for 1 hour. Then, acetonitrile (140 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and obtained the dimer oligonucleotide precursor DMTrO-T(H)T-SUC-TOB (727 mg, 98% yield).

[0551] Example 3: Oxidation using compound (Ic-1) Under an argon atmosphere, the oligonucleotide precursor obtained in Synthesis Example 2 (171 mg, 93 μmol), dichloromethane (4.7 mL), and acetonitrile (1.4 mL) were added to a 50 mL two-necked flask to prepare a solution. Subsequently, water (60 μL), compound (Ic-1) (43 mg, 189 μmol), and diazabicycloundecene (DBU) (55 μL, 372 μmol) were added, and the solution was stirred at room temperature for 2 hours. After that, acetonitrile (42 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and quantitatively obtained the oligonucleotide DMTrO-T(OH)T-SUC-TOB as a dimer. m / z:Calcd. 1843.16,Found 1842.15[MH] -

[0552] Example 4: Oxidation using compound (Ib-1) Under an argon atmosphere, the oligonucleotide precursor obtained in Synthesis Example 2 (170 mg, 93 μmol), dichloromethane (4.7 mL), and acetonitrile (1.4 mL) were added to a 50 mL two-necked flask to prepare a solution. Subsequently, compound (Ib-1) (62 mg, 186 μmol) and DBU (55 μL, 372 μmol) were added, and the solution was stirred at room temperature for 2 hours. After that, acetonitrile (42 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and quantitatively obtained the oligonucleotide DMTrO-T(OH)T-SUC-TOB as a dimer. m / z:Calcd. 1843.16,Found 1842.15[MH] -

[0553] Experimental Example 1 Non-patent document 1 (ARKIVOC 2009 (iii) 264-273) discloses that a dinucleotide (compound 4) can be produced by condensing and oxidizing an H-phosphonate (compound 1) and a nucleoside (compound 2) in the presence of a large amount of triphenylphosphine and a large amount of 2,2'-dipyridyl disulfide, as shown in the following formula.

[0554] [ka]

[0555] Furthermore, repeated extension during oligonucleotide synthesis can unintentionally remove the protecting group (e.g., 2-cyanoethyl group) from the thiophosphate ester portion of the oligonucleotide. Therefore, oligonucleotides from which the protecting group of the thiophosphate ester portion has been removed were reacted with 2,2'-dipyridyl disulfide used in Non-Patent Literature 1 or compound (Ib-1) used in the present invention to evaluate 2,2'-dipyridyl disulfide and compound (Ib-1). In the following experiments, a large amount of 2,2'-dipyridyl disulfide or compound (Ib-1) was used relative to the substrate (i.e., oligonucleotide from which the protecting group of the thiophosphate ester portion has been removed) in accordance with the reaction conditions described in Non-Patent Literature 1.

[0556] (1) Experiments using 2,2'-dipyridyl disulfide and triphenylphosphine A mixture of HO-T(S)T(S)T-SUC-TOB and Piv-TOB from which the 2-cyanoethyl group has been removed (40 mg: 20 mg as HO-T(S)T(S)T-SUC-TOB, 0.012 mmol) was dissolved in anhydrous dichloromethane (0.3 mL) and anhydrous pyridine (0.3 mL) to prepare a solution. Triphenylphosphine (15 mg, 0.059 mmol) and 2,2'-dipyridyl disulfide (25 mg, 0.12 mmol) were added, and the solution was stirred at room temperature for 4.5 hours. Then, acetonitrile (7 mL) was added to the solution, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, followed by drying under reduced pressure to obtain a solid (31.2 mg).

[0557] The obtained solid (10 mg) and 28 wt% aqueous ammonia (5 mL) were placed in an autoclave and heated at 65°C for 4 hours, then cooled to room temperature. After removing insoluble matter from the reaction mixture using a syringe filter, the mixture was concentrated under reduced pressure using a centrifugal evaporator, and the resulting concentrate was analyzed by LC-MS.

[0558] (2) Experiments using compound (Ib-1) and water A mixture of HO-T(S)T(S)T-SUC-TOB and Piv-TOB from which the cyanoethyl group had been removed (40 mg: 20 mg as HO-T(S)T(S)T-SUC-TOB, 0.012 mmol) was dissolved in anhydrous dichloromethane (0.3 mL) and anhydrous pyridine (0.3 mL) to prepare a solution. Water (6 μL, 0.3 mmol) and compound (Ib-1) (39 mg, 0.12 mmol) were added, and the solution was stirred at room temperature for 4.5 hours. Subsequently, acetonitrile (7 mL) was added to the solution, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, followed by vacuum drying to quantitatively obtain the solid.

[0559] The obtained solid (10 mg) and 28 wt% aqueous ammonia (5 mL) were placed in an autoclave and heated at 65°C for 4 hours, then cooled to room temperature. After removing insoluble matter from the reaction mixture using a syringe filter, the mixture was concentrated under reduced pressure using a centrifugal evaporator, and the resulting concentrate was analyzed by LC-MS.

[0560] (3) Evaluation The amount of oligonucleotide HO-T(S)T(S)T-OH (hereinafter referred to as "the target oligonucleotide") having a thiophosphate ester bond, and the amount of by-products (hereinafter referred to as "by-products") formed by the reaction of the target oligonucleotide with an oxidizing agent (2,2'-dipyridyl disulfide or compound (Ib-1)) were measured as follows, and the proportion of by-products was calculated. Specifically, the concentrate obtained in the above experiment was analyzed by LC-MS, and the peak area of ​​each compound (target oligonucleotide and by-product) was calculated using the extracted ion chromatogram (EIC) of each observed compound, and the following formula was used: Percentage of by-products (%) = (Peak area of ​​by-products / (Peak area of ​​target oligonucleotide + Peak area of ​​by-products) × 100) The proportion of by-products was calculated using the following method. The results are shown in Table 2.

[0561] [Table 2]

[0562] As shown in Table 2, the use of 2,2'-dipyridyl disulfide and triphenylphosphine resulted in the formation of a large amount of byproducts. On the other hand, no byproducts were detected even when a large amount of compound (Ib-1) and water were used. Furthermore, the method described in Non-Patent Document 1, which uses 2,2'-dipyridyl disulfide and triphenylphosphine, also results in the formation of a large amount of byproducts, similar to the results shown in this Experimental Example 1.

[0563] Synthesis Example 3: Synthesis of 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)) In a 200 mL round-bottom flask, tetrahydrofuran (100 mL), 2-mercaptopropionic acid (10 mmol), and compound (Ib-1) (20 mmol) were added, and the solution was stirred for 24 hours. The solution was then filtered, and the solvent from the filtrate was removed under reduced pressure. The resulting concentrate was purified by silica gel column chromatography to obtain the target product (1.6 g). 1 H-NMR(400MHz, methanol-d4)δ 7.91(d,J=8.0,1H),7.81(d,J=8.0,1H),7.47(dd,J=8.0,1H),7.38(dd,J=8.0,1H),3.89(q,J=7.1,1H),1.58(d,J=7.1,3H)

[0564] Synthesis Example 4: Synthesis of 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)) A solution was prepared by adding tetrahydrofuran (100 mL), 3-mercaptopropionic acid (10 mmol), and compound (Ib-1) (20 mmol) to a 200 mL round-bottom flask, and the solution was stirred for 24 hours. The solution was then filtered, and the solvent from the filtrate was removed under reduced pressure. The resulting concentrate was purified by silica gel column chromatography to obtain the target product (1.3 g). 1 H-NMR(400MHz,chloroform-d)δ 7.93(d,J=8.0,1H),7.81(d,J=8.0,1H),7.48(dd,J=8.0,1H),7.39(dd,J=8.0,1H),3.24(t,J=6.8,2H),2.81(t,J=6.8,2H).

[0565] Synthesis Example 5: Synthesis of 2,2'-dibenzoimidazolyl disulfide (compound (Ib-4)) Dichloromethane (150 mL), 2-mercaptobenzimidazole (20 mmol), and triethylamine (22 mmol) were added to a 300 mL round-bottom flask, followed by the addition of iodine (11 mmol). The solution was stirred for 3 hours. The solution was then filtered, and the solvent from the filtrate was removed under reduced pressure. The resulting concentrate was recrystallized using diethyl ether and ethanol to obtain the target product (2.5 g). 1 H-NMR (400MHz, DMSO-d 6 )δ=7.58-7.56(m,4H),7.26-7.22(m,4H).

[0566] Synthesis Example 6: Synthesis of 2,2'-dibenzoxazolyl disulfide (compound (Ib-5)) A solution was prepared by adding 2-mercaptobenzoxazole (1.5 g, 10 mmol) and ethyl acetate (18 mL) to a 30 mL round-bottom flask. Hydrogen peroxide (33 wt%, 489 μL) was added dropwise to the solution over 60 minutes at 0°C, and the solution was then heated to room temperature and stirred for 2.5 hours. Water (1.4 mL) was added to the solution, and the mixture was stirred for 10 minutes. The organic layer was separated, and the aqueous layer was extracted twice with ethyl acetate (5 mL). The solution obtained from the extraction was dried over sodium sulfate, and the solvent was removed under reduced pressure. The obtained solid was washed with ice-cold diethyl ether and then recrystallized with diethyl ether. The precipitated solid was collected by filtration, dried under reduced pressure, and the target product (210 mg) was obtained. 1 H-NMR(400MHz,Chloroform-d)δ=7.76-7.69(m,2H),7.58-7.50(m,2H),7.42-7.32(m,4H).

[0567] Synthesis Example 7: Synthesis of 2,2'-dithiobis(pyridine-N-oxide) (compound (Ib-8)) A solution was prepared by adding 2-mercaptopyridine-N-oxide (3.8 g, 30 mmol) and water (27 mL) to a 100 mL round-bottom flask. Hydrogen peroxide (33 wt%, 3.2 mL) was added dropwise to the solution over 60 minutes at 0°C, and the solution was then heated to 40°C and stirred for 1 hour. The solution was filtered to collect the solid, which was washed with methanol and dried under reduced pressure to obtain the target product (1.2 g). 1 H-NMR (400MHz, Chloroform-d) δ=8.31(d,J=5.2,2H),7.62(d,J=6.4,2H),7.34-7.24(m,2H),7.20(dd,J=6.4,2H).

[0568] Synthesis Example 8: Synthesis of an oligonucleotide precursor containing a phosphite ester bond Under an argon atmosphere, HO-T-SUC-TOB (2.1 g, 1.7 mmol), anhydrous dichloromethane (87 mL), and anhydrous acetonitrile (26 mL) were added to a 200 mL two-necked flask to prepare the solution. Then, molecular sieve 3A (1.7 g), triphenylphosphine (227 mg, 867 μmol), dT-CE phosphoramidite (2.6 g, 3.5 mmol), and 5-ethylthio-1H-tetrazole (452 ​​mg, 3.47 mmol) were added sequentially, and the mixture was stirred at room temperature for 60 minutes. Trifluoroethanol (1.3 mL, 17 mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 30 minutes. Then, 2,6-xylidine (2.6 mL, 21 mmol) and DDTT (892 mg, 4.34 mmol) were added, and the solution was stirred at room temperature for 60 minutes. After removing molecular sieve 3A by filtration, indole (2.0 g, 17 mmol) and trifluoroacetic acid (4.8 mL, 63 mmol) were added sequentially, and the solution was stirred at room temperature for 30 minutes. Further addition of trifluoroacetic acid (265 μL, 3.46 mmol) was made, and the solution was stirred at room temperature for 30 minutes. Subsequently, pyridine (16 mL, 200 mmol) and water (742 μL) were added sequentially, and the solution was stirred at room temperature for 60 minutes. Acetonitrile (858 mL) was then added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and obtained the dimer oligonucleotide HO-T(S)T-SUC-TOB (2.6 g, yield 94%). m / z:Calcd.1610.03,Found 1611.05[M+H] +

[0569] Under an argon atmosphere, HO-T(S)T-SUC-TOB (1.3 g, 811 μmol), anhydrous dichloromethane (40 mL), and anhydrous acetonitrile (12 mL) were added to a 100 mL two-necked flask to prepare a solution. Then, molecular sieve 3A (816 mg), triphenylphosphine (109 mg, 417 μmol), dC(Bz)-CE phosphoramidite (1.4 g, 1.6 mmol), and 5-ethylthio-1H-tetrazole (212 mg, 1.63 μmol) were added sequentially, and the solution was stirred at room temperature for 60 minutes. After removing molecular sieve 3A by filtration, acetonitrile (30 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and obtained the oligonucleotide precursor DMTrO-C(III)T(S)T-SUC-TOB, a trimer (1.8 g, 96% yield). m / z:Calcd. 2342.26,Found 2343.28[M+H] +

[0570] Examples 5-15 and Comparative Example 2: Oxidation using various oxidizing agents A solution was prepared by adding DMTrO-C(III)T(S)T-SUC-TOB (60 mg, 25 μmol), anhydrous dichloromethane (1.3 mL), and anhydrous acetonitrile (0.4 mL) to a flask. After adding water (17 μL) to the solution, various oxidizing agents (38 μmol) shown in Table 3 below were added, and oxidation was carried out by stirring at room temperature for 3 hours to synthesize the triper oligonucleotide DMTrO-C(O)T(S)T-SUC-TOB (hereinafter referred to as "oxidized product"). In Examples 6 and 7, where 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)) or 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)) was used as the oxidizing agent, as shown in Table 3 below, in addition to oxidation in the presence of water as described above, oxidation was also carried out under anhydrous conditions without adding water to the solution in the above procedure.

[0571] Compounds (Ib-2) to (Ib-5) and (Ib-8) were those obtained in the aforementioned synthesis examples 3 to 7. Compound (Ic-1) used was synthesized by the method described in the Supporting Information of Non-Patent Document 2. Other oxidizing agents used were commercially available products.

[0572] Comparison of Examples 5-15 and Comparative Example 2 The oxidized solutions (10 μL) of Examples 5-15 and Comparative Example 2, along with DDTT (1 mg), were placed in a 1 mL vial. After dilution with tetrahydrofuran (450 μL), DBU (20 μL) was added, and the solution was stirred at room temperature for 30 seconds to sulfurize the unreacted DMTrO-C(III)T(S)T-SUC-TOB, synthesizing DMTrO-C(S)T(S)T-SUC-TOB (hereinafter referred to as "sulfide"), and a test solution containing the oxidized and sulfide was prepared. MS analysis of the obtained test solution was performed, and using the abundance of each compound observed (oxidized (i.e., the target oligonucleotide DMTrO-C(O)T(S)T-SUC-TOB) and sulfide), the following formula was derived: Percentage of oxidized form (%) = (Abundance of oxidized form / (Abundance of oxidized form + Abundance of sulfide form)) × 100 The proportion of oxidized products was calculated using the following method. The results are shown in Table 3. The numbers in parentheses in Examples 6 and 7 of Table 3 indicate the proportion of oxidized products when oxidation was carried out under anhydrous conditions.

[0573] [Table 3]

[0574] As shown in Table 3, by using compounds (Ib-1) to (Ib-10) or (Ic-1) as oxidizing agents, oligonucleotide precursors can be oxidized more efficiently than 2,2'-dipyridyl disulfide used in Non-Patent Document 1 (ARKIVOC 2009 (iii) 264-273).

[0575] Example 16: Synthesis of DMTrO-Af(O)Cm(O)T(O)T-OH using compound (Ib-1) as an oxidizing agent Under an argon atmosphere, HO-T-SUC-TOB (351 mg, 284 μmol), Piv-TOB (350 mg, 350 μmol), anhydrous dichloromethane (14 mL), and anhydrous acetonitrile (4 mL) were added to a 300 mL two-necked flask to prepare the solution. Then, dT-CE phosphoramidite (402 mg, 540 μmol) and 5-ethylthio-1H-tetrazole (69.9 mg, 537 μmol) were added sequentially, and the solution was stirred at room temperature for 45 minutes. Furthermore, dT-CE phosphoramidite (21 mg, 29 μmol) and 5-ethylthio-1H-tetrazole (3.7 mg, 28 μmol) were added, and the solution was stirred at room temperature for 35 minutes. Then, water (184 μL, 10.2 mmol) was added, and the solution was stirred at room temperature for 30 minutes. Compound (Ib-1) (282 mg, 848 μmol) was added to the solution, and the solution was stirred at room temperature for 60 minutes. Then, triphenylphosphine (74.2 mg, 283 μmol) was added, and the solution was stirred for 30 minutes. Subsequently, 2,3-dimethylfuran (298 μL, 2.83 mmol) and trifluoroacetic acid (563 μL, 7.35 mmol) were added sequentially, and the solution was stirred at room temperature for 30 minutes. Furthermore, trifluoroacetic acid (86.6 μL, 1.13 mmol) was added, and the solution was stirred at room temperature for 30 minutes. Subsequently, pyridine (2.0 mL, 25 mmol) and water (120 μL) were added sequentially to the solution, and the solution was stirred at room temperature for 60 minutes. Subsequently, acetonitrile (170 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel. The mixture was then dried under reduced pressure to obtain a mixture of Piv-TOB and its dimer, oligonucleotide HO-T(O)T-SUC-TOB (766 mg, yield 96%). m / z:Calcd. 1594.06,Found 1595.07[M+H] +

[0576] A mixture of Piv-TOB and the triper oligonucleotide HO-Cm(O)T(O)T-SUC-TOB was obtained in the same manner as above, except that the solid obtained above (766 mg) was used and 2'-OMe-C-CE phosphoramidite was used instead of dT-CE phosphoramidite (875 mg, yield 98%). m / z:Calcd.2070.17,Found 2071.19[M+H] +

[0577] Under an argon atmosphere, the solid obtained above (875 mg), anhydrous dichloromethane (13 mL), and anhydrous acetonitrile (4 mL) were added to a 200 mL two-necked flask to prepare a solution. Then, 2'-FA-CE phosphoramidite (441 mg, 503 μmol) and 5-ethylthio-1H-tetrazole (65.7 mg, 505 μmol) were added sequentially, and the solution was stirred at room temperature for 45 minutes. Furthermore, 2'-FA-CE phosphoramidite (25 mg, 28 μmol) and 5-ethylthio-1H-tetrazole (3.4 mg, 26 μmol) were added, and the solution was stirred at room temperature for 60 minutes. Next, water (172 μL, 9.54 mmol) was added, and the solution was stirred at room temperature for 60 minutes. Then, compound (Ib-1) (266 mg, 800 μmol) was added to the solution, and the solution was stirred at room temperature for 60 minutes. After that, triphenylphosphine (71.1 mg, 271 μmol) was added, and the solution was stirred at room temperature for 30 minutes. Subsequently, acetonitrile (129 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, and then dried under reduced pressure to obtain a mixture of Piv-TOB and the tetramer oligonucleotide DMTrO-Af(O)Cm(O)T(O)T-SUC-TOB (1007 mg, yield 93%). m / z:Calcd.2860.41,Found 2861.42[M+H] +

[0578] The solid obtained above (9.3 mg) and 28 wt% aqueous ammonia (5 mL) were placed in an autoclave and heated at 65°C for 4 hours, then cooled to room temperature. After removing insoluble matter from the reaction mixture using a syringe filter, the mixture was concentrated under reduced pressure using a centrifugal evaporator to obtain an aqueous solution of DMTrO-Af(O)Cm(O)T(O)T-OH. m / z:Calcd. 1498.39,Found 1497.37[MH] -

[0579] Example 17: Synthesis of DMTrO-Af(O)Cm(O)T(O)T-OH using compound (Ic-1) as an oxidizing agent An aqueous solution of DMTrO-Af(O)Cm(O)T(O)T-OH was obtained in the same manner as in Example 16, except that compound (Ib-1) was replaced with compound (Ic-1). m / z:Calcd. 1498.39,Found 1497.37[MH] -

[0580] Comparative Example 3: Synthesis of DMTrO-Af(O)Cm(O)T(O)T-OH using iodine as an oxidizing agent Under an argon atmosphere, HO-T-SUC-TOB (354 mg, 286 μmol) and Piv-TOB (351 mg, 352 μmol) were added to a 200 mL two-necked flask, followed by the addition of anhydrous dichloromethane (14 mL) and anhydrous acetonitrile (4.2 mL) to prepare the solution. Subsequently, dT-CE phosphoramidite (406 mg, 545 μmol) and 5-ethylthio-1H-tetrazole (70.1 mg, 539 μmol) were added sequentially, and the solution was stirred at room temperature for 45 minutes. Furthermore, dT-CE phosphoramidite (23 mg, 31 μmol) and 5-ethylthio-1H-tetrazole (4.0 mg, 31 μmol) were added, and the solution was stirred at room temperature for 35 minutes. Next, water (180 μL, 10.2 mmol) was added, and the solution was stirred at room temperature for 30 minutes. Then, pyridine (91 μL, 1.1 mmol) and iodine (145 mg, 569 μmol) were added sequentially, and the solution was stirred at room temperature for 90 minutes. Subsequently, 2,3-dimethylfuran (298 μL, 2.83 mmol) and trifluoroacetic acid (650 μL, 8.48 mmol) were added sequentially, and the solution was stirred at room temperature for 30 minutes. Further, trifluoroacetic acid (86.6 μL, 1.13 mmol) was added, and the solution was stirred at room temperature for 30 minutes. Then, pyridine (2.3 mL, 29 mmol) and water (120 μL) were added sequentially, and the solution was stirred at room temperature for 60 minutes. Subsequently, acetonitrile (140 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel. After drying under reduced pressure, a mixture of Piv-TOB and its dimer, oligonucleotide HO-T(O)T-SUC-TOB, was quantitatively obtained (827 mg). m / z:Calcd.1594.06,Found 1595.07[M+H] +

[0581] A mixture of Piv-TOB and the triper oligonucleotide HO-Cm(O)T(O)T-SUC-TOB was obtained using the same procedure as above, except that the solid obtained above (812 mg) was replaced with 2'-OMe-C-CE phosphoramidite. (893 mg, yield 94%) m / z:Calcd.2070.17,Found 2071.19[M+H] +

[0582] Under an argon atmosphere, the solid obtained above (889 mg), anhydrous dichloromethane (13 mL), and anhydrous acetonitrile (4.0 mL) were added to a 200 mL two-necked flask to prepare the solution. Then, 2'-FA-CE phosphoramidite (729 mg, 833 μmol) and 5-ethylthio-1H-tetrazole (107 mg, 819 μmol) were added sequentially, and the solution was stirred at room temperature for 105 minutes. Subsequently, water (174 μL, 9.63 mmol) was added, and the solution was stirred at room temperature for 30 minutes. Then, pyridine (200 μL, 2.48 mmol) and iodine (317 mg, 1.25 mmol) were added sequentially, and the solution was stirred at room temperature for 60 minutes. Finally, triphenylphosphine (109 mg, 414 μmol) was added, and the solution was stirred at room temperature for 30 minutes. Subsequently, acetonitrile (137 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel. After drying under reduced pressure, a mixture of Piv-TOB and the tetramer oligonucleotide DMTrO-Af(O)Cm(O)T(O)T-SUC-TOB was obtained (1052 mg, yield 95%). m / z:Calcd. 2860.41,Found 2861.42[M+H] +

[0583] The solid obtained above (9.8 mg) and 28 wt% aqueous ammonia (5 mL) were placed in an autoclave and heated at 65°C for 4 hours, then cooled to room temperature. After removing insoluble matter from the reaction mixture using a syringe filter, the mixture was concentrated under reduced pressure using a centrifugal evaporator to obtain an aqueous solution of DMTrO-Af(O)Cm(O)T(O)T-OH. m / z:Calcd. 1498.39,Found 1497.37[MH] -

[0584] Comparison of Examples 16, 17 and Comparative Example 3 DMTrO-Af(O)Cm(O)T(O)T-OH (hereinafter referred to as "the target oligonucleotide") in aqueous solutions obtained in Examples 16, 17, and Comparative Example 3, and trimers DMTrO-Af(O)Cm(O)T-OH or DMTrO-Af(O)T(O)T-OH (hereinafter referred to as "the defective compound"), which are obtained by deleting the 2nd or 3rd residue of the target oligonucleotide, respectively, were measured by MS analysis. For MS analysis, the peak area of ​​each compound was calculated using the extracted ion chromatogram (EIC) of each observed compound (the target oligonucleotide and the defective compound), and the following formula was used: Percentage of defective cells (%) = (Sum of peak areas of each defective cell / Peak area of ​​the target oligonucleotide) × 100 The percentage of deficient cells was calculated using the following method. The results are shown in Table 4.

[0585] [Table 4]

[0586] As is clear from the results in Table 4, the formation of deficiencies can be suppressed by using compound (Ib-1) or compound (Ic-1) instead of iodine as the oxidizing agent.

[0587] Example 18: Synthesis of HO-Af(O)Cm(O)T(O)T-SUC-TOB using compound (Ic-1) as an oxidizing agent (with addition of aromatic amine) Under an argon atmosphere, HO-T-SUC-TOB (6.4 g, 5.2 mmol), Piv-TOB (13 g, 13 mmol), anhydrous dichloromethane (349 mL), and anhydrous acetonitrile (98 mL) were added to a 3 L three-necked flask to prepare the solution. Then, dT-CE phosphoramidite (7.3 g, 9.8 mmol) and 5-ethylthio-1H-tetrazole (1.3 g, 9.8 mmol) were added sequentially, and the solution was stirred at room temperature for 65 minutes. Furthermore, dT-CE phosphoramidite (385 mg, 517 μmol) and 5-ethylthio-1H-tetrazole (67.3 mg, 517 μmol) were added, and the solution was stirred at room temperature for 30 minutes. After that, water (932 μL, 51.7 mmol) was added, and the solution was stirred at room temperature for 40 minutes. Next, compound (Ic-1) (3.5 g, 15 mmol) was added, and the solution was stirred for 60 minutes. Triphenylphosphine (1.4 g, 5.2 mmol) and 2,6-xylidine (5.5 mL, 45 mmol) as an aromatic amine were sequentially added to the solution, and the solution was stirred at room temperature for 30 minutes. Subsequently, 2,3-dimethylfuran (5.4 mL, 52 mmol) and trifluoroacetic acid (13.8 mL, 180 mmol) were sequentially added, and the solution was stirred at room temperature for 80 minutes. Then, pyridine (43.7 mL, 541 mmol) and water (2.5 mL) were sequentially added to the solution, and the solution was stirred at room temperature for 90 minutes. After that, acetonitrile was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and a mixture of Piv-TOB and the dimer oligonucleotide HO-T(O)T-SUC-TOB was obtained (21 g, yield 98%). m / z:Calcd. 1594.06,Found 1595.07[M+H] +

[0588] A mixture of Piv-TOB and the trimer oligonucleotide HO-Cm(O)T(O)T-SUC-TOB was quantitatively obtained (23 g) in the same manner as above, except that the solid obtained above (21 g) was used and 2'-OMe-C-CE phosphoramidite was used instead of dT-CE phosphoramidite. m / z:Calcd. 2070.17,Found 2071.19[M+H] +

[0589] A mixture of Piv-TOB and the tetramer oligonucleotide HO-Af(O)Cm(O)T(O)T-SUC-TOB was obtained (23 g, 99% yield) in the same manner as above, except that the solid obtained above (21 g) was used and 2'-FA-CE phosphoramidite was used instead of 2'-OMe-C-CE phosphoramidite. m / z:Calcd. 2558.28,Found 2559.29[M+H] +

[0590] Furthermore, to confirm the effect of adding aromatic amines, 1.3 mL of the reaction solution was withdrawn from the reaction solution before the addition of 2,3-dimethylfuran, and acetonitrile (9 mL) was added to the solution. The precipitated solid was filtered by suction using a Kiriyama funnel and then dried to obtain a mixture of Piv-TOB and the tetramer oligonucleotide DMTrO-Af(O)Cm(O)T(O)T-SUC-TOB (69 mg). m / z:Calcd. 2860.41,Found 2861.42[M+H] +

[0591] The DMTrO-Af(O)Cm(O)T(O)T-SUC-TOB (3.4 mg) and 28 wt% aqueous ammonia (5 mL) obtained above were placed in an autoclave and heated at 65°C for 4 hours, then cooled to room temperature. After removing insoluble matter from the reaction mixture using a syringe filter, the mixture was concentrated under reduced pressure using a centrifugal evaporator to obtain an aqueous solution of DMTrO-Af(O)Cm(O)T(O)T-OH. m / z:Calcd.1498.39,Found 1497.37[MH] -

[0592] Comparison of Example 17 and Example 18 DMTrO-Af(O)Cm(O)T(O)T-OH (hereinafter referred to as "the target oligonucleotide") obtained in aqueous solutions in Examples 17 and 18, and trimers DMTrO-Af(O)Cm(O)T-OH or DMTrO-Af(O)T(O)T-OH (hereinafter referred to as "the defective compound"), which are obtained by deleting the 2nd or 3rd residue of the target oligonucleotide, respectively, were measured by MS analysis. For MS analysis, the peak are...

Claims

1. A method for producing an oligonucleotide having a phosphate ester bond by oxidizing an oligonucleotide precursor having a phosphite ester bond or a phosphonic acid ester bond with an oxidizing agent, The oxidizing agent is defined by formula (I): 【Chemistry 1】 [In the formula, X １ This consists of a single bond, a sulfur atom, an oxygen atom, and -S (=O). ２ - or -N (-R ３ ) - indicates, X １ If it is a single bond, １ R indicates a halogen atom, and ２ This represents an optionally substituted aryl group or an optionally substituted heteroaryl group. X １ is a sulfur atom or -S(=O) ２ -, when R １ represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R ２ represents an optionally substituted aryl group or an optionally substituted heteroaryl group, X １ If R is an oxygen atom, １ and R ２ Together with the sulfur and oxygen atoms to which they are bonded, they form a heterocycle which may be substituted, and X １ ga-N(-R ３ ) - If R １ R represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R ２ and R ３ They, together with the sulfur and nitrogen atoms to which they bond, form a heterocycle which may be substituted, or R １ and R ３ Together with the nitrogen atoms to which they are bonded, they form a heterocycle that may be substituted, and R ２ This indicates an optionally substituted aryl group or an optionally substituted heteroaryl group. A method using a compound represented by (excluding 2,2-dipyridyl disulfide).

2. The compound represented by formula (I) is formula (Ia): 【Chemistry 2】 [In the formula, R １ａ This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Ring A represents a 5- or 6-membered unsaturated heterocycle. Y １ａ is -S(=O)-, -S(=O) ２ -, -C(=O)- or -C(=S)-, m represents an integer between 0 and 3. m R ２ａ Each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and If m is an integer greater than or equal to 2, then two adjacent R ２ａ It may form a fused ring, which may be substituted, together with ring A. Compounds represented by formula (Ib): 【Transformation 3】 [In the formula, R １ｂ This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Ring B represents a 6-membered aromatic hydrocarbon ring, or a 5- or 6-membered aromatic heterocycle. n represents an integer from 0 to 5. n R ２ｂ Each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and If n is an integer greater than or equal to 2, then two adjacent R ２ｂ Together with ring B, it may form a substituted, bicyclic fused aromatic heterocycle. Compounds represented by (excluding 2,2-dipyridyl disulfide) The method according to claim 1.

3. The compound represented by formula (Ia) is given by formula (Ic): 【Chemistry 4】 [In the formula, R １ｃ This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Y １ｃ is -S(=O)-, -S(=O) ２ -, -C(=O)- or -C(=S)-, p represents 0 or 1. R ２ｃ This represents a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. Ring C represents a 6-membered aromatic hydrocarbon ring, a 6-membered nitrogen-containing aromatic heterocycle, or a 10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom. q represents an integer from 0 to 4, and q R ３ｃ Each of these independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. The method according to claim 2, wherein the compound is represented by the compound shown in the claim.

4. The compound represented by formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazole Lu-3(2H)-one, 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one, 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole- 3(2H)-one, 2-phenylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one, 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-phenyl-2H-1,2-benzothiadin-3(4H)-one, 2-(2,6-dimethylphenyl)isothiazolo[5,4 The method according to claim 3, wherein at least one is selected from the group consisting of -b]pyridine-3(2H)-one, 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one, 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one, and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one.

5. The method according to claim 3, wherein the ring C is a six-membered aromatic hydrocarbon ring or a six-membered nitrogen-containing aromatic heterocycle.

6. The compound represented by formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazole-3(2H)-one, 2-(2,6-dimethyl The method according to claim 5, wherein at least one is selected from the group consisting of phenyl)-5-nitro-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one, and 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3(2H)-one.

7. Formula (r1) in the compound represented by the above formula (Ib): 【Transformation 5】 The base represented by, (1) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (2) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (4) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (6) A (pyridine-N-oxide)-2-yl group which may be substituted with one to four substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (7) A 4-pyridyl group which may be substituted with one to four substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (8) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (9) A phenyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (10) A 2-imidazolyl group which may be substituted with one to three substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. The method according to any one of claims 2 to 6.

8. The method according to claim 7, wherein the compound represented by formula (Ib) is at least one selected from the group consisting of 2,2'-dibenzothiazolyl disulfide, 2-(2-benzothiazolyldithio)propanoic acid, 3-(2-benzothiazolyldithio)propanoic acid, 2,2'-dibenzimidazolyl disulfide, 2,2'-dibenzoxazolyl disulfide, 5,5'-di(1,2,3-triazolyl)disulfide, 5,5'-dithiobis(1-phenyl-1H-tetrazole), 2,2'-dithiobis(pyridine-N-oxide), 4,4'-dipyridyl disulfide, 3-(2-pyridyldithio)propanoic acid, 2,2'-dithiobis(1H-imidazole), and 2,2'-dithiobis(1-methyl-1H-imidazole).

9. The method according to any one of claims 1 to 8, wherein the oxidation is carried out in the presence of water.

10. Oligonucleotide precursors having phosphite ester bonds or phosphonic acid ester bonds, (1) Oligonucleotide precursors having phosphite ester bonds obtained by condensation of a nucleoside, nucleotide, or oligonucleotide with a phosphoramidite-treated nucleoside, nucleotide, or oligonucleotide, or (2) Oligonucleotide precursors having phosphite bonds obtained by condensation of oligonucleotides with phosphoramidites The method according to any one of claims 1 to 9.

11. The method according to any one of claims 1 to 10, wherein the oligonucleotide precursor having a phosphite ester bond or a phosphonic acid ester bond is an oligonucleotide precursor having a thiophosphate ester bond in addition to the phosphite ester bond or phosphonic acid ester bond.

12. A method for producing oligonucleotides by one-pot synthesis, The aforementioned one-pot synthesis, Step (1) involves condensing a nucleoside, nucleotide, or oligonucleotide (a) having a hydrophobic protecting group with a nucleoside, nucleotide, or oligonucleotide (b) that is protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions and is phosphoramidized, in a solution containing a nonpolar solvent, to form an oligonucleotide precursor (c) having a phosphite ester bond and a hydrophobic protecting group, and protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions. Step (2) involves adding a quenching agent (i) for the phosphoramidized nucleoside, nucleotide, or oligonucleotide (b) to the solution after step (1) to quench the phosphoramidized nucleoside, nucleotide, or oligonucleotide (b). Step (3) involves adding an oxidizing agent to the solution after step (2) to oxidize the oligonucleotide precursor (c) to form an oligonucleotide (d) having a phosphate ester bond and a hydrophobic protecting group, and being protected by a temporary protecting group whose hydroxyl group can be removed under acidic conditions. Step (3) involves adding an oxidizing agent quenching agent (ii) to the solution after step (4) to quench the oxidizing agent. Step (5) involves adding an acid to the solution after step (4) to remove the temporary protecting group of oligonucleotide (d) that can be removed under acidic conditions, thereby forming oligonucleotide (e) having an unprotected hydroxyl group and a hydrophobic protecting group, and Step (7) to add a polar solvent to the solution containing the oligonucleotide (e) to precipitate the oligonucleotide (e). Including, and The oxidizing agent is defined by formula (I): 【Transformation 6】 [In the formula, X １ This consists of a single bond, a sulfur atom, an oxygen atom, and -S (=O). ２ - or -N (-R ３ ) - indicates, X １ If it is a single bond, １ R indicates a halogen atom, and ２ This represents an optionally substituted aryl group or an optionally substituted heteroaryl group. X １ is a sulfur atom or -S (=O) ２ - If R １ R represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and ２ This represents an optionally substituted aryl group or an optionally substituted heteroaryl group. X １ If R is an oxygen atom, １ and R ２ Together with the sulfur and oxygen atoms to which they are bonded, they form a heterocycle which may be substituted, and X １ ga-N(-R ３ ) - If R １ R represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R ２ and R ３ They, together with the sulfur and nitrogen atoms to which they bond, form a heterocycle which may be substituted, or R １ and R ３ Together with the nitrogen atoms to which they are bonded, they form a heterocycle that may be substituted, and R ２ This indicates an optionally substituted aryl group or an optionally substituted heteroaryl group. A method using a compound represented by (excluding 2,2-dipyridyl disulfide).

13. The compound represented by formula (I) is formula (Ia): 【Transformation 7】 [In the formula, R １ａ This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Ring A represents a 5- or 6-membered unsaturated heterocycle. Y １ａ is -S(=O)-, -S(=O) ２ -, -C(=O)- or -C(=S)-, m represents an integer between 0 and 3. m R ２ａ Each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and If m is an integer greater than or equal to 2, then two adjacent R ２ａ It may form a fused ring, which may be substituted, together with ring A. Compounds represented by formula (Ib): 【Transformation 8】 [In the formula, R １ｂ This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Ring B represents a 6-membered aromatic hydrocarbon ring, or a 5- or 6-membered aromatic heterocycle. n represents an integer from 0 to 5. n R ２ｂ Each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and If n is an integer greater than or equal to 2, then two adjacent R ２ｂ Together with ring B, it may form a substituted, bicyclic fused aromatic heterocycle. Compounds represented by (excluding 2,2-dipyridyl disulfide) The method according to claim 12.

14. The compound represented by formula (Ia) is given by formula (Ic): 【Chemistry 9】 [In the formula, R １ｃ This represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. Y １ｃ is -S(=O)-, -S(=O) ２ -, -C(=O)- or -C(=S)-, p represents 0 or 1. R ２ｃ This represents a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. Ring C represents a 6-membered aromatic hydrocarbon ring, a 6-membered nitrogen-containing aromatic heterocycle, or a 10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom. q represents an integer from 0 to 4, and q R ３ｃ Each of these independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group. The method according to claim 13, wherein the compound is represented by the compound.

15. The compound represented by formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazole Lu-3(2H)-one, 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one, 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3 (2H)-one, 2-phenylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one, 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-phenyl-2H-1,2-benzothiadin-3(4H)-one, 2-(2,6-dimethylphenyl)isothiazolo[5,4- The method according to claim 14, wherein at least one is selected from the group consisting of [b]pyridine-3(2H)-one, 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one, 2-(4-methoxyphenyl)isothiazolo[5,4-b]pyridine-3(2H)-one, and 2-(4-fluorophenyl)isothiazolo[5,4-b]pyridine-3(2H)-one.

16. The method according to claim 14, wherein the ring C is a six-membered aromatic hydrocarbon ring or a six-membered nitrogen-containing aromatic heterocycle.

17. The compound represented by formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridine-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazole-3(2H)-one, 2-(2,6-dimethyl The method according to claim 16, wherein at least one is selected from the group consisting of phenyl)-5-nitro-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazole-3(2H)-one, 5-nitro-2-(4-pyridyl)-1,2-benzothiazole-3(2H)-one, and 2-[1-methyl-1-(2-pyridyl)ethyl]-5-nitro-1,2-benzothiazole-3(2H)-one.

18. Formula (r1) in the compound represented by the above formula (Ib): 【Chemistry 10】 The base represented by, (1) A 2-benzothiazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (2) A 2-benzimidazolyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (3) A 2-benzoxazolyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (4) A 5-(1,2,3-triazolyl) group which may be substituted with one or two substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups, (5) A 1H-tetrazole-5-yl group which may be substituted with one substituent selected from the group consisting of optionally substituted alkyl groups and optionally substituted aryl groups, (6) A (pyridine-N-oxide)-2-yl group which may be substituted with one to four substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. (7) A 4-pyridyl group which may be substituted with one to four substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (8) A 2-pyridyl group which may be substituted with 1 to 4 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, (9) A phenyl group which may be substituted with 1 to 5 substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups, or (10) A 2-imidazolyl group which may be substituted with one to three substituents selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups, and electron-withdrawing groups. The method according to any one of claims 13 to 17.

19. The method according to claim 18, wherein the compound represented by formula (Ib) is at least one selected from the group consisting of 2,2'-dibenzothiazolyl disulfide, 2-(2-benzothiazolyldithio)propanoic acid, 3-(2-benzothiazolyldithio)propanoic acid, 2,2'-dibenzimidazolyl disulfide, 2,2'-dibenzoxazolyl disulfide, 5,5'-di(1,2,3-triazolyl)disulfide, 5,5'-dithiobis(1-phenyl-1H-tetrazole), 2,2'-dithiobis(pyridine-N-oxide), 4,4'-dipyridyl disulfide, 3-(2-pyridyldithio)propanoic acid, 2,2'-dithiobis(1H-imidazole), and 2,2'-dithiobis(1-methyl-1H-imidazole).

20. The method according to any one of claims 12 to 19, wherein the quenching agent (i) is water.

21. The method according to any one of claims 12 to 20, wherein the quenching agent (ii) is an organophosphorus compound.

22. The method according to claim 21, wherein the organophosphorus compound is at least one selected from the group consisting of phosphines, phosphite triesters, phosphiniteroates, phosphonic acid diesters, and phosphiniteroates.

23. The method according to claim 21, wherein the organophosphorus compound is a phosphine.

24. The method according to any one of claims 12 to 23, wherein the one-pot synthesis further comprises adding aromatic amines to the solution after step (3).

25. The method according to any one of claims 12 to 24, wherein the one-pot synthesis further comprises a step (6) of adding a base to the solution after step (5) after step (5) and before step (7).

26. The method according to any one of claims 12 to 25, wherein the oligonucleotide precursor (c) is an oligonucleotide precursor having a thiophosphate ester bond in addition to a phosphite ester bond.

27. Formula (In): 【Chemistry 11】 [In the formula, (1) R １ｎ represents an optionally substituted phenyl group, p' represents 0, ring C'' represents a 10-membered bicyclic fused aromatic heterocycle containing a nitrogen atom, and q' represents 0 or (2) R １ｎ represents an optional phenyl group, p' represents 0, ring C'' represents a 6-membered nitrogen-containing aromatic heterocycle, q' represents 1, and R ３ｎ is C １－６ Does it indicate a perfluoroalkyl group? (3) Caution １ｎ represents an optionally substituted phenyl group, p' represents 1, ring C'' represents a 6-membered nitrogen-containing aromatic heterocycle, and q' represents 0, or (4) R １ｎ is one C １－６ This indicates a pyridyl group substituted with an alkyl group, where p' represents 0, the ring C'' represents a 6-membered nitrogen-containing aromatic heterocycle, and q' represents 0. A compound represented by the formula.

28. The compound according to claim 27, wherein the compound represented by formula (In) is 2-phenylisothiazolo[5,4-b]quinoline-3(2H)-one, 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridine-3(2H)-one, 2-phenyl-2H-1,2-benzothiadin-3(4H)-one, or 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridine-3(2H)-one.

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