Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
  • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
  • C07D513/04—Ortho-condensed systems
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D279/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one sulfur atom as the only ring hetero atoms
  • C07D279/02—1,2-Thiazines; Hydrogenated 1,2-thiazines
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H1/00—Processes for the preparation of sugar derivatives
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
  • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
  • C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
  • Y02P20/00—Technologies relating to chemical industry
  • Y02P20/50—Improvements relating to the production of bulk chemicals
  • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

• the present invention relates to a method for producing oligonucleotides.
• the phosphoramidite method and the H-phosphonate method are widely used as methods for producing oligonucleotides.
• a phosphoramidite (compound (1)) and a nucleoside (compound (2)) are condensed to have a phosphite ester bond.
• An oligonucleotide (compound (4)) is obtained by synthesizing an oligonucleotide precursor (compound (3)) and then oxidizing the oligonucleotide precursor.
• the "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.
• DMTr represents a 4,4'-dimethoxytrityl group
• i-Pr represents an isopropyl group
• CE represents a 2-cyanoethyl group
• Base and Base' represent an optionally protected nucleobase.
• R represents a protecting group or solid support.
• nucleosides instead of nucleosides, nucleotides or oligonucleotides (for example, oligonucleotides having a thiophosphate ester bond) may be condensed. Further, in the phosphoramidite method, instead of oxidizing the oligonucleotide precursor, sulfuration of the oligonucleotide precursor may be performed.
• H-phosphonate (compound (5)) and pivaloyl chloride (PivCl) are reacted to form an activated H-phosphonate (compound ( 6)), condensing it with a nucleoside (compound (7)) to synthesize an oligonucleotide precursor having a phosphonate ester bond (compound (8)), and then oxidizing the oligonucleotide precursor An oligonucleotide (compound (9)) is obtained by doing so.
• DMTr represents 4,4′-dimethoxytrityl
• N(Et) 3 represents triethylamine
• Base and Base′ represent an optionally protected nucleobase
• Piv represents a pivaloyl group
• R represents a protecting group or solid support.
• nucleosides instead of nucleosides, nucleotides or oligonucleotides (for example, oligonucleotides with thiophosphate ester bonds) may be condensed. Also, in the H-phosphonate method, instead of oxidizing the oligonucleotide precursor, sulfuration of the oligonucleotide precursor may be performed.
• H-phosphonate (compound 1) and nucleoside (compound 2) are synthesized in the presence of a large amount of triphenylphosphine and a large amount of 2,2'-dipyridyl disulfide, as shown by the following formula. is disclosed to condense and oxidize to produce a dinucleotide (compound 4).
• DMTr represents 4,4′-dimethoxytrityl
• Th represents a thymine base
• PPh 3 represents a triphenylphosphine
• Ac represents an acetyl group.
• Non-Patent Document 2 and Patent Document 1 do not disclose a method for producing an oligonucleotide
• benzoisothiazolone is produced by triethyl phosphite (P(OEt) 3 ) as represented by the following formula. ) derivative (compound 1) (in other words, oxidation of triethyl phosphite (P(OEt) 3 ) by the benzisothiazolone derivative (compound 1) to give triethyl phosphate (O ⁇ P(OEt) 3 ).
• forming is disclosed.
• iodine is widely used as an oxidizing agent for oxidizing an oligonucleotide precursor having a phosphite ester bond (for example, the compound represented by formula (3) above).
• iodine is widely used as an oxidizing agent for oxidizing an oligonucleotide precursor having a phosphonate ester bond (for example, the compound represented by the above formula (8)).
• the present inventors have found that when iodine is used as an oxidizing agent for oligonucleotide precursors with phosphite linkages, the phosphite linkage sites of the precursors interact with the oxidizing agent to decompose. It was found that a by-product formed by In addition to phosphite or phosphonate linkages, oxidation of oligonucleotide precursors having thiophosphate linkages with iodine converts the thiophosphate linkages to phosphate linkages, forming It was found that a by-product (hereinafter sometimes referred to as "desulfurized product”) is generated.
• the present invention has been made in view of the circumstances described above, and an object of the present invention is to provide an oligonucleotide capable of suppressing the formation of the above-mentioned deletion product or desulfurization product compared to the case of using iodine. It is to provide a manufacturing method.
• 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 comprising:
• the oxidizing agent has the formula (I):
• R 1a 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 heterocyclic ring
• m represents an integer of 0 to 3
• m R 2a each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and when m is an integer of 2 or more, adjacent Two R 2a 's may form an optionally substituted condensed ring together with ring A.
• R 1b 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 of 0 to 5
• Each of the n R 2b independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and when n is an integer of 2 or more, adjacent Two R 2b 's may form an optionally substituted bicyclic condensed aromatic heterocyclic ring together with ring B.
• a compound represented by (excluding 2,2-dipyridyl disulfide) The method according to [1] above.
• R 1c represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group
• p represents 0 or 1
• R 2c 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 heterocyclic ring, or a 10-membered bicyclic condensed aromatic heterocyclic ring containing a nitrogen atom
• q represents an integer of 0 to 4
• q R 3c each independently represent an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group.
• the compound represented by the formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridin-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine- 3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridin-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazol-3(2H)-one, 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazol-3(2H)-one , 5-nitro-2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one, 5-nitro-2-(4-pyridyl)-1,2-benzothiazol-3(2H) -one, 2-[1-methyl-1-(2-pyridyl)ethyl
• the compound represented by formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridin-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine- 3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridin-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazol-3(2H)-one, 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazol-3(2H)-one , 5-nitro-2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazol-3(2H
• the compound represented by the formula (Ib) is 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).
• an oligonucleotide precursor having a phosphite ester bond or a phosphonate ester bond (1) an oligonucleotide precursor having a phosphite ester bond obtained by condensation of a nucleoside, nucleotide or oligonucleotide with a phosphoramidized nucleoside, nucleotide or oligonucleotide; or (2) an oligonucleotide and , which is an oligonucleotide precursor having a phosphite ester bond obtained by condensation with a phosphoramidite, the method according to any one of [1] to [9].
• a method for producing an oligonucleotide by one-pot synthesis comprising: The one-pot synthesis is In a solution containing a non-polar solvent, a nucleoside, nucleotide or oligonucleotide (a) having a hydrophobic protecting group, a hydroxyl group protected with a temporary protecting group removable under acidic conditions, and phosphoramidated Further, an oligo which is condensed with a nucleoside, nucleotide or oligonucleotide (b) and has a phosphite ester bond and a hydrophobic protecting group and the hydroxyl group is protected by a temporary protecting group removable under acidic conditions step (1) of forming a nucleotide precursor (c); A quenching agent (i) for the phosphoramidated nucleoside, nucleotide or oligonucleotide (b) is added to the solution after step (1) to quench the phosphoramidated nucleoside,
• step (3) of forming an oligonucleotide (d) protected with a temporary protecting group step (4) of adding a quenching agent (ii) for the oxidizing agent to the solution after step (3) to quench the oxidizing agent; adding an acid to the solution after step (4) to remove the temporary protecting group removable under acidic conditions of the oligonucleotide (d), resulting in an oligonucleotide having an unprotected hydroxyl group and a hydrophobic protecting group;
• R 1a 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 heterocyclic ring
• m represents an integer of 0 to 3
• m R 2a each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and when m is an integer of 2 or more, adjacent Two R 2a 's may form an optionally substituted condensed ring together with ring A.
• R 1b 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 of 0 to 5
• Each of the n R 2b independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, and when n is an integer of 2 or more, adjacent Two R 2b 's may form an optionally substituted bicyclic condensed aromatic heterocyclic ring together with ring B.
• a compound represented by (excluding 2,2-dipyridyl disulfide) The method according to [12] above.
• R 1c represents an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group
• p represents 0 or 1
• R 2c 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 heterocyclic ring, or a 10-membered bicyclic condensed aromatic heterocyclic ring containing a nitrogen atom
• q represents an integer of 0 to 4
• q R 3c each independently represent an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group.
• the compound represented by the formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridin-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine- 3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridin-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazol-3(2H)-one, 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazol-3(2H)-one , 5-nitro-2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one, 5-nitro-2-(4-pyridyl)-1,2-benzothiazol-3(2H) -one, 2-[1-methyl-1-(2-pyridyl)ethyl
• the compound represented by formula (Ic) is 2-phenylisothiazolo[5,4-b]pyridin-3(2H)-one, 2-isopropylisothiazolo[5,4-b]pyridine- 3(2H)-one, 2-isopropylisothiazolo[4,5-c]pyridin-3(2H)-one, 2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one, 5-nitro-2-phenyl-1,2-benzothiazol-3(2H)-one, 2-(2,6-dimethylphenyl)-5-nitro-1,2-benzothiazol-3(2H)-one , 5-nitro-2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one, 5-nitro-2-(2-pyridyl)-1,2-benzothiazol-3(2H
• the compound represented by the formula (Ib) is 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).
• the quenching agent (ii) is an organophosphorus compound.
• the organic phosphorus compound is at least one selected from the group consisting of phosphines, phosphite triesters, phosphite esters, phosphonite diesters and phosphinate esters. .
• the organic phosphorus compound is a phosphine.
• the one-pot synthesis further comprises step (6) of adding a base to the solution after step (5) after step (5) and before step (7), above [12] to [24 ]
• step (6) of adding a base to the solution after step (5) after step (5) and before step (7), above [12] to [24 ] The method according to any one of
• oligonucleotide precursor (c) is an oligonucleotide precursor having a thiophosphate ester bond in addition to a phosphite ester bond the method of.
• R 1n represents an optionally substituted phenyl group
• p′ represents 0, ring C′′ represents a 10-membered bicyclic condensed aromatic heterocycle containing a nitrogen atom
• q ' indicates 0, or
• R 1n represents an optionally substituted phenyl group
• p′ represents 0, ring C′′ represents a 6-membered nitrogen-containing aromatic heterocycle
• q′ represents 1
• R 3n represents a C 1-6 perfluoroalkyl group
• R 1n represents an optionally substituted phenyl group
• p′ represents 1, ring C′′ represents a 6-membered nitrogen-containing aromatic heterocycle
• q′ represents 0, or
• R 1n represents a pyridyl group substituted with one C 1-6 alkyl group
• p′ represents 0, ring C′′ represents a 6-membered nitrogen-containing aromatic heterocyclic ring
• q ' indicates 0.
• the compound represented by the formula (In) is 2-phenylisothiazolo[5,4-b]quinolin-3(2H)-one, 2-phenyl-5-trifluoromethylisothiazolo[5, 4-b]pyridin-3(2H)-one, 2-phenyl-2H-1,2-benzothiazin-3(4H)-one, or 2-(4-methyl-2-pyridinyl)isothiazolo[5,4- b]
• the compound according to the above [27] which is pyridin-3(2H)-one.
• oligonucleotides by suppressing the production of the above-mentioned deletion products or desulfurization products compared to the case of using iodine.
• C ab means having a carbon number of a or more and b or less (a and b are integers).
• halogen atom means a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.
• hydrocarbon group includes, for example, an aliphatic hydrocarbon group, an araliphatic hydrocarbon group, a monocyclic saturated hydrocarbon group, an aromatic hydrocarbon group, and the like. includes monovalent groups and divalent groups such as alkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, aryl groups, aralkyl groups and alkylene groups.
• alkyl (group) may be linear or branched.
• Alkyl (group) includes an alkyl group having 1 or more carbon atoms, preferably a C 1-10 alkyl group, more preferably a C 1-6 if there is no particular limitation on the carbon number range. is an alkyl group. Specific examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl and the like.
• alkenyl (group) may be linear or branched.
• Alkenyl (group) includes, for example, a C 2-6 alkenyl group and the like. Specific examples include vinyl, 1-propenyl, allyl, isopropenyl, butenyl, isobutenyl and the like.
• alkynyl (group) may be linear or branched.
• Alkynyl (group) includes, for example, a C 2-6 alkynyl group and the like. Specific examples 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.
• cycloalkyl (group) means a cyclic alkyl group, and includes, for example, a C 3-8 cycloalkyl group, preferably a C 3-6 cycloalkyl group. Specific examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.
• aryl (group) means a monocyclic or polycyclic (condensed) aromatic hydrocarbon group, such as phenyl, 1-naphthyl, 2-naphthyl, biphenylyl, 1-anthryl and C 6-14 aryl groups such as , 2-anthryl and the like. Among them, a C 6-10 aryl group is more preferred, and phenyl is particularly preferred.
• heteroaryl refers to a monocyclic or polycyclic (condensed) aromatic ring containing heteroatoms selected from nitrogen, sulfur and oxygen atoms in addition to carbon atoms. group heterocyclic group.
• Heteroaryl groups include, for example, 5- or 6-membered monocyclic heteroaryl groups and 8- to 14-membered fused polycyclic heteroaryl groups.
• heteroaryl includes, for example, thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, (pyridine-N- oxid)-yl, pyrazinyl, pyrimidinyl, pyridazinyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, triazolyl, tetrazolyl, triazinyl, etc. mentioned.
• the "heteroaryl group” includes a (pyridin-N-oxide)-yl group represented by the following formula (where * indicates the bonding position).
• the "5- or 6-membered monocyclic nitrogen-containing heteroaryl (group)” includes, for example, the above-mentioned “5- or 6-membered monocyclic heteroaryl (group)” Examples include those containing at least one or more nitrogen atoms as atoms.
• 8- to 14-membered condensed polycyclic heteroaryl includes, for example, benzothiophenyl, benzofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzisothiazolyl, ryl, benzotriazolyl, imidazopyridinyl, thienopyridinyl, furopyridinyl, pyrrolopyridinyl, pyrazolopyridinyl, oxazolopyridinyl, thiazolopyridinyl, imidazopyrazinyl, imidazopyrimidinyl, thienopyrimidinyl, furopyrimidinyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl, oxazolopyrimidinyl, thiazolopyrimidinyl, pyrazolotriazinyl,
• the "8- to 14-membered condensed polycyclic nitrogen-containing heteroaryl (group)” includes, for example, the aforementioned “8- to 14-membered condensed polycyclic heteroaryl (group)", Examples include those containing at least one or more nitrogen atoms as ring-constituting atoms.
• aralkyl (group) includes C 7-20 aralkyl groups, preferably C 7-16 aralkyl groups (C 6-10 aryl-C 1-6 alkyl groups). Specific examples thereof include benzyl, 1-phenylethyl, 2-phenylethyl, 1-phenylpropyl, naphthylmethyl, 1-naphthylethyl, 1-naphthylpropyl and the like.
• alkylene (group) may be linear or branched.
• the “alkylene (group)” includes an alkylene group having 1 or more carbon atoms, preferably a C 1-10 alkylene group, more preferably a C 1-6 alkylene group, unless the carbon number range is particularly limited. It is an alkylene group. Specific examples include methylene, ethylene, propylene, butylene, pentylene and hexylene.
• alkoxy (group) may be linear or branched.
• Alkoxy (group) includes an alkoxy group having 1 or more carbon atoms, preferably a C 1-10 alkoxy group, more preferably a C 1-6 alkoxy group when there is no particular limitation on the carbon number range. It is an alkoxy group. Specific examples thereof include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy and the like.
• acyl (group) may be linear or branched.
• Examples of “acyl (group)” include C 1-6 alkanoyl group, C 7-13 aroyl group and the like.
• Specific examples of “acyl (group)” include formyl, acetyl, n-propionyl, isopropionyl, n-butyryl, isobutyryl, pivaloyl, valeryl, hexanoyl, benzoyl, naphthoyl, levulinyl and the like.
• “Acyl (group)” may be substituted.
• the term "6-membered aromatic hydrocarbon ring” means a benzene ring.
• the "6-membered aromatic hydrocarbon ring” forms part of the condensed ring
• the "6-membered aromatic hydrocarbon ring” means a benzene ring forming part of the condensed ring. do.
• 5- or 6-membered aromatic heterocyclic ring includes, for example, thiophene ring, furan ring, 1H-pyrrole ring, imidazole ring, pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, 1,2,4-oxadiazole ring, 1,3,4-oxadiazole ring, 1,2,4-thiadiazole ring, 1,3,4-thiadiazole ring, triazole ring, tetrazole ring, pyridine ring , pyridine-N-oxide ring, pyrazine ring, pyrimidine ring, pyridazine ring, triazine ring and the like.
• aromatic heterocycle includes pyridine-N-oxide ring.
• the "4- to 8-membered nitrogen-containing heterocyclic ring” includes, for example, azetidine ring, pyrrolidine ring, pyrroline ring, 2H-pyrrole ring, piperidine ring, piperazine ring, azacycloheptane ring, azacyclooctane ring , 1H-pyrrole ring, imidazole ring, pyrazole ring, thiazole ring, isothiazole ring, oxazole ring, isoxazole ring, 1,2,4-oxadiazole ring, 1,3,4-oxadiazole ring, 1, 2,4-thiadiazole ring, 1,3,4-thiadiazole ring, triazole ring, tetrazole ring, pyridine ring, pyridine-N-oxide ring, pyrazine ring, pyrimidine ring, pyridazine ring, tria
• the "5- or 6-membered nitrogen-containing heterocyclic ring” includes, for example, the above-mentioned “4- to 8-membered nitrogen-containing heterocyclic ring” having 5 or 6 ring-constituting atoms. be done.
• the "6-membered nitrogen-containing aromatic heterocyclic ring” includes, for example, pyridine ring, pyridine-N-oxide ring, pyrazine ring, pyrimidine ring, pyridazine ring, triazine ring and the like.
• the "bicyclic condensed nitrogen-containing heterocyclic ring” includes, for example, pyrrolidine ring, decahydroisoquinoline ring, decahydroquinoline ring, benzothiophene ring, benzofuran ring, isobenzofuran ring, benzimidazole ring, benzo oxazole ring, benzisoxazole ring, benzothiazole ring, benzisothiazole ring, benzotriazole ring, indole ring, isoindole ring, 1H-indazole ring, purine ring, isoquinoline ring, 5,6,7,8-tetrahydro isoquinoline ring, quinoline ring, 5,6,7,8-tetrahydroquinoline ring, phthalazine ring, pteridine ring, naphthyridine ring, quinoxaline ring, quinazoline ring, cinn
• the "bicyclic condensed aromatic heterocyclic ring” includes, for example, a benzothiophene ring, a benzofuran ring, an isobenzofuran ring, a benzimidazole ring, a benzoxazole ring, a benzisoxazole ring, a benzothiazole ring, a benzo isothiazole ring, benzotriazole ring, indole ring, isoindole ring, 1H-indazole ring, purine ring, isoquinoline ring, 5,6,7,8-tetrahydroisoquinoline ring, quinoline ring, 5,6,7,8 -tetrahydroquinoline ring, phthalazine ring, pteridine ring, naphthyridine ring, quinoxaline ring, quinazoline ring, cinnoline ring and the like.
• the "bicyclic condensed aromatic heterocycle” includes one ring that is an aromatic heterocycle and the other ring that is a non-aromatic heterocyclic ring or a non-aromatic hydrocarbon ring. Also included are certain bicyclic fused rings (eg, 5,6,7,8-tetrahydroisoquinoline rings).
• the "nitrogen atom-containing 10-membered bicyclic condensed aromatic heterocycle” includes, for example, 5,6,7,8-tetrahydroisoquinoline ring, quinoline ring and the like.
• a perchloroalkyl group e.g., trichlor
• a "perchloroalkyl group” means an alkyl group in which all hydrogen atoms are substituted with chlorine atoms.
• perfluoroalkyl group means an alkyl group in which all hydrogen atoms are substituted with fluorine atoms.
• the "substituent” includes, for example, the above-mentioned halogen atom, alkyl group, aralkyl group, alkoxy group, acyl group, alkenyl group, alkynyl group, cycloalkyl group, aryl group, hydroxyl group, nitro cyano group, guanidyl group, carboxy group, alkoxycarbonyl group (the alkoxy moiety is the same as the alkoxy group), sulfo group, phospho group, alkylsulfanyl group (the alkyl moiety is the same as the alkyl group), alkylsulfinyl group (alkyl part is the same as the alkyl group), alkylsulfonyl group (the alkyl part is the same as the alkyl group), amino group, monoalkylamino group (the alkyl part is the same as the alkyl group), dialkylamino group (the alkyl part is the
• the "hydroxyl-protecting group” is not particularly limited, and known protecting groups can be used.
• the protecting group 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-N
• the "temporary protecting group for a hydroxyl group removable under acidic conditions” includes, for example, a trityl group, a 9-phenyl-9-xanthenyl group, a 9-phenyl-9-thioxanthenyl group, a 1,1-bis (4-Methoxyphenyl) -1-phenylmethyl group (herein sometimes referred to as “4,4'-dimethoxytrityl group”) and other bis (C 1-6 alkoxy) trityl groups, 1-( and mono(C 1-18 alkoxy)trityl groups such as 4-methoxyphenyl)-1,1-diphenylmethyl group (which may be referred to as “4-monomethoxytrityl group” in this specification).
• the temporary protective group is preferably 4,4'-dimethoxytrityl group or 4-monomethoxytrityl group, more preferably 4,4'-dimethoxytr
• the "amino-protecting group” is not particularly limited, and known protecting groups can be used.
• the protecting group include pivaloyl group, pivaloyloxymethyl group, acetyl group, trifluoroacetyl group, phenoxyacetyl group, 4-isopropylphenoxyacetyl group, 4-tert-butylphenoxyacetyl group, benzoyl group and isobutyryl group.
• (2-hexyl)decanoyl group dimethylformamidinyl group, 1-(dimethylamino)ethylidene group, and 9-fluorenylmethyloxycarbonyl group.
• protecting group for a phosphate group is not particularly limited, and known protecting groups can be used.
• Phosphate protecting groups include, for example: 2-cyanoethyl group represented by the following formula (Pg-1), 2-[2-(4,4′-dimethoxytrityloxy)ethylsulfonyl]ethyl group represented by the following formula (Pg-2), [3-(4,4′-dimethoxytrityloxy)-2,2-di(ethoxycarbonyl)]propyl group represented by the following formula (Pg-3), [3-(4,4'-dimethoxytrityloxy)-2,2-di(N-methylcarbamoyl)]propyl group represented by the following formula (Pg-4).
• the phosphate group-protecting group is preferably a 2-cyanoethyl group.
• Me represents a methyl group
• Et represents an ethyl group
• DMTr represents a 4,4'-dimethoxytrityl group
• * represents a bonding position with a phosphate group.
• hydrophobic protecting group for example, a protecting group described as “anchor” in WO 2017/104836 can be used.
• the hydrophobic protecting group is preferably a protecting group represented by the following formula (Pg-5): **L-Y-Z (Pg-5) [In the formula, ** indicates the position of attachment to the group to be protected; L is a single bond, or formula (Pg-i-1) or (Pg-i-2):
• R 1p and R 2p each independently represent a C 1-22 hydrocarbon group
• L 1 represents an optionally substituted divalent C 1-22 hydrocarbon group, and —CH 2 — in the divalent C 1-22 hydrocarbon group may be replaced with a linker
• R 4p represents a C 1-22 alkylene group
• R 3p and R 5p each independently represent a hydrogen atom or a C 1 -22 alkyl group, or R 3p and R 5p together may form a ring).
• Y represents a single bond, an oxygen atom, or NR (R represents a hydrogen atom, an alkyl group or an aralkyl group); and Z represents formulas (Pg-ii-1) to formulas (Pg-ii-5 ) either:
• Ring A' represents a benzene ring
• Ring B' represents a cyclohexane ring
• Ring D' represents a naphthalene ring or a bicyclic condensed aromatic heterocycle
• R 6p is a hydrogen atom, or when R b is a group represented by the following formula ( Pg -iii), R 6p of ring A' or ring B' is may represent a single bond or -O- to form a condensed ring together with ring A' or ring B' and ring C'
• k represents an integer of 1 to 4
• Ring C' represents a benzene ring: j represents an integer from 0 to 4; j Q are each independently as defined above; j R 9p each independently represents a hydrocarbon group in which a linear aliphatic hydrocarbon group having 10 or more carbon atoms is bonded via a single bond or a linker; R 8p represents a hydrogen atom, or represents a single bond or -O- together with R 6p of ring A' or ring B' to form a condensed ring together with ring A' or ring B' and ring C' may form Ring C' is, in addition to j QR 9p , a halogen atom, a C 1-6 alkyl group optionally substituted with a halogen atom, and a C 1-6 alkoxy group optionally substituted with a halogen atom It may have a substituent selected from the group consisting of ), or R a and R b together form an oxo group;
• ring A', ring C', R6p and R8p form a condensed ring
• ring A' and ring C' each represent a benzene ring portion in the condensed ring.
• ring B', ring C', R 6p and R 8p form a condensed ring
• ring C' represents a benzene ring moiety in the condensed ring
• ring B' represents a cyclohexane ring moiety in the condensed ring.
• the linear aliphatic hydrocarbon group having 10 or more carbon atoms is preferably a group selected from a linear C 10-40 alkyl group and a linear C 10-40 alkenyl group, more preferably linear C 10-40 alkyl group, more preferably linear C 10-30 alkyl group, particularly preferably linear C 12-28 alkyl group, most preferably linear is a C 14-26 alkyl group.
• 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 to 3 benzyl group in which a linear C 10-40 alkyl group is bonded via —O—, or cyclohexylmethyl in which 1 to 3 linear C 10-40 alkyl groups are bonded via —O— is the base.
• L 1 is a divalent C 1-22 hydrocarbon group; and L 2 is a single bond.
• L 1 is an ethylene group
• L 1 is an ethylene group
• L 1 is an ethylene group
• L 1 is an optionally substituted phenylene group
• L represented by formula (Pg-i-1) is particularly preferably a succinyl group.
• L represented by formula (Pg-i-2) in formula (Pg-5) will be described.
• L 1 in formula (Pg-i-2) is preferably a divalent C 6-10 aromatic hydrocarbon group, more preferably a phenylene group.
• L 2 in formula (Pg-i-2) is preferably a single bond.
• a preferred combination of L 1 and L 2 in formula (Pg-i-2) is a combination in which L 1 is a divalent C 6-10 aromatic hydrocarbon group and L 2 is a single bond.
• a more preferred combination of L 1 and L 2 in formula (Pg-i-2) is a combination in which L 1 is a phenylene group and L 2 is a single bond.
• R 1p and R 2p in formula (Pg-i-2) are each independently preferably a C 1-22 alkyl group, more preferably a C 1-10 alkyl group.
• a preferred embodiment of L represented by formula (Pg-i-2) is R 1p and R 2p are each independently a C 1-22 alkyl group; L 1 is a divalent C 6-10 aromatic hydrocarbon group; and L 2 is a group that is a single bond.
• R 1p and R 2p are each independently a C 1-10 alkyl group; L 1 is a phenylene group; and L 2 is a group that is a single bond.
• R is preferably a hydrogen atom, a C 1-6 alkyl group or a C 7-16 aralkyl group, more preferably a hydrogen atom, methyl, ethyl or It is benzyl, more preferably a hydrogen atom.
• Y is preferably a single bond, an oxygen atom or NR, more preferably a single bond or an oxygen atom.
• R 6p is preferably a hydrogen atom.
• R a and R b are preferably hydrogen atoms or together form an oxo group.
• a preferred embodiment of Z represented by formula (Pg-ii-1) is R a and R b are hydrogen atoms; R 6p is a hydrogen atom; k is an integer from 1 to 3; the k Q's are —O—; and the k R 7p 's are each independently a linear C 10-40 alkyl group, is the base.
• R a and R b are hydrogen atoms;
• R 6p is a hydrogen atom;
• k is an integer from 1 to 3;
• k Q are —O—;
• k R 7p are each independently a benzyl group to which 1 to 3 linear C 10-40 alkyl groups are bonded via —O— , or a cyclohexylmethyl group in which 1 to 3 linear C 10-40 alkyl groups are linked via —O—, is the base.
• R a is a hydrogen atom
• R 6p is a hydrogen atom
• k is an integer from 1 to 3
• k R 7p are each independently a linear C 10-40 alkyl group
• R b is of formula (Pg-iii) (formula in which * is a binding position, j is an integer of 0 to 3, j Qs are -O-, and j R 9p are each independently a linear C 10- 40 is an alkyl group
• R 8p is a hydrogen atom. is the base.
• R a is a hydrogen atom
• k is an integer from 1 to 3
• k R 7p are each independently a linear C 10-40 alkyl group
• R b is of formula (Pg-iii) (formula in which * is a binding position, j is an integer of 0 to 3, j Qs are -O-, and j R 9p are each independently a linear C 10- 40 alkyl group, and R 8p represents a single bond or -O- together with R 6p to form a condensed ring with ring A' and ring C'.)
• R 8p represents a single bond or -O- together with R 6p to form a condensed ring with ring A' and ring C'.
• Z represented by formula (Pg-ii-1) is R a and R b together form an oxo group; R 6p is a hydrogen atom; k is an integer from 1 to 3; the k Q's are —O—; and the k R 7p 's are each independently a linear C 10-40 alkyl group, is the base.
• Z represented by formula (Pg-ii-1) is R a and R b together form an oxo group;
• R 6p is a hydrogen atom;
• k is an integer from 1 to 3;
• k number of Q is -O-;
• k R 7p are each independently a benzyl group in which 1 to 3 linear C 10-40 alkyl groups are bonded via —O—, or 1 to 3 linear C 10 -40 alkyl group is a cyclohexylmethyl group bonded via -O-, is the base.
• a preferred embodiment of Z represented by formula (Pg-ii-2) is R 6p is a hydrogen atom; k is an integer from 1 to 3; k Q are —O—; and k R 7p are each independently a linear C 10-40 alkyl group, 1 to 3 linear C 10-40 alkyl groups is a benzyl group bonded via -O-, or a cyclohexylmethyl group bonded via -O- with 1 to 3 linear C 10-40 alkyl groups, is the base.
• R 6p is preferably a hydrogen atom.
• R a and R b are preferably hydrogen atoms or together form an oxo group.
• a preferred embodiment of Z represented by formula (Pg-ii-3) is R a and R b are hydrogen atoms; R 6p is a hydrogen atom; k is an integer from 1 to 3; the k Q's are —O—; and the k R 7p 's are each independently a linear C 10-40 alkyl group, is the base.
• R a and R b are hydrogen atoms;
• R 6p is a hydrogen atom;
• k is an integer from 1 to 3;
• k Q are —O—;
• k R 7p are each independently a benzyl group to which 1 to 3 linear C 10-40 alkyl groups are bonded via —O— , or a cyclohexylmethyl group in which 1 to 3 linear C 10-40 alkyl groups are linked via —O—, is the base.
• R a is a hydrogen atom
• R 6p is a hydrogen atom
• k is an integer from 1 to 3
• k R 7p are each independently a linear C 10-40 alkyl group
• R b is of formula (Pg-iii) (formula in which * is a binding position, j is an integer of 0 to 3, j Qs are -O-, and j R 9p are each independently a linear C 10- 40 is an alkyl group
• R 8p is a hydrogen atom. is the base.
• R a is a hydrogen atom; k is an integer from 1 to 3; and k R 7p are each independently a linear C 10-40 alkyl group; and R b is of formula (Pg-iii) (formula in which * is a binding position, j is an integer of 0 to 3, j Qs are -O-, and j R 9p are each independently a linear C 10- 40 alkyl group, and R 8p represents a single bond or -O- together with R 6p to form a condensed ring with ring B' and ring C'). , is the base.
• R a and R b together form an oxo group
• R 6p is a hydrogen atom
• k is an integer from 1 to 3
• the k Q's are —O—
• the k R 7p 's are each independently a linear C 10-40 alkyl group, is the base.
• R a and R b together form an oxo group
• R 6p is a hydrogen atom
• k is an integer from 1 to 3
• k number of Q is -O-
• k R 7p are each independently a benzyl group in which 1 to 3 linear C 10-40 alkyl groups are bonded via —O—, or 1 to 3 linear C 10 -40 alkyl group is a cyclohexylmethyl group bonded via -O-, is the base.
• a preferred embodiment of Z represented by formula (Pg-ii-4) is Ring D' is a naphthalene ring; R a and R b are hydrogen atoms; k is an integer from 1 to 3; k Q′ are —O—; and k R 7p are each independently a linear C 10-40 alkyl group, 1 to 3 linear C 10-40 alkyl the group is a benzyl group linked via -O-, or a cyclohexylmethyl group linked via -O- with 1 to 3 linear C 10-40 alkyl groups, is the base.
• Z represented by formula (Pg-ii-4) is ring D' is an indole ring; R a and R b are hydrogen atoms; k is 1; Q′ is a single bond; and R 7p is a linear C 10-40 alkyl group, benzyl in which 1 to 3 linear C 10-40 alkyl groups are bonded via —O— or a cyclohexylmethyl group to which 1 to 3 linear C 10-40 alkyl groups are attached via —O—; and R 7p is a group attached to the nitrogen atom of the indole ring be.
• a preferred embodiment of Z represented by formula (Pg-ii-5) is u is 1 or 2; and R 10p is a C 4-10 perfluoroalkyl group.
• Z is preferably a group represented by formula (Pg-ii-1), formula (Pg-ii-2) or formula (Pg-ii-3).
• the protecting group (Pg-5) is preferably L is a succinyl group or a group represented by formula (Pg-i-2) (wherein R 1p and R 2p are each independently C 1-10 alkyl L 1 is a divalent phenylene group, and L 2 is a single bond.), and YZ is a 3,4,5-tris(octadecyloxy)benzyloxy group, 3,5-bis(docosyloxy)benzyloxy group, 3,5-bis[3′,4′,5′-tris(octadecyloxy)benzyloxy]benzyloxy group, 3,4,5-tris[3′, 4′,5′-tris(octadecyloxy)benzyloxy]benzyloxy group, 3,4,5-tris(octadecyloxy)benzylamino group, 2,4-bis(docosyloxy)benzylamino group, 3,5-bis (docosyloxy)benzylamino group
• 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, 3,5-bis[3′, 4′,5′-tris(octadecyloxy)benzyloxy]benzyloxy group, 3,4,5-tris[3′,4′,5′-tris(octadecyloxy)benzyloxy]benzyloxy group, 3,4 , 5-tris(octadecyloxy)benzylamino group, 2,4-bis(docosyloxy)benzylamino group, 3,5-bis(docosyloxy)benzylamino group, bis(4-docosyloxyphenyl)methylamino group, 4 -Methoxy-2-[3',4',5'-tris(octadecyloxy
• 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, 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)
• the protecting group (Pg-5) is particularly 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 or a phenyl(2,3 ,4-tris(octadecyloxy)phenyl)methylamino group, or LY is a succinyl-1,4-piperazinediyl group, and Z is 3,4,5-tris( octadecyloxy)benzoyl group.
• the protecting group (Pg-5) is most preferably L is a succinyl group, and YZ is a 3,4,5-tris(octadecyloxy)benzyloxy group or a 3,4,5-tris(octadecyloxy)cyclohexylmethyloxy group or LY is a succinyl-1,4-piperazinediyl group and Z is a 3,4,5-tris(octadecyloxy)benzoyl group.
• the protecting group (Pg-5), and the compounds used to form it, are prepared by known methods (e.g., WO2017/104836, WO2019/131719, WO2020/235658, It can be formed or manufactured by a method described in Japanese Patent Publication No. 2021/039935 or International Publication No. 2021/198883) or a method analogous thereto.
• solid-phase support is not particularly limited as long as it is used for solid-phase synthesis of oligonucleotide synthesis in the relevant field, and examples thereof include glass beads, resin beads, and the like. mentioned.
• the support or resin used as a solid phase carrier can be any support or resin known in the art and suitable for use in solid phase synthesis.
• Said supports or resins include, for example, polystyrene supports (which may be further functionalized with, for example, p-methylbenzyl-hydrylamine), diatomaceous earth-encapsulated polydimethylacrylamide (pepsin K), silica, microparticles, Examples include porous glass, amphipathic polystyrene-polyethylene glycol (PEG) resin, PEG-polyamide, PEG-polyester resin, Wang-PEG resin, Rink-amide PEG resin, and the like. Said support or resin may be modified. Modified supports or resins include, for example, long chain alkylamino Controlled Pore Glass (lcaa CPG) and the like.
• nucleoside which is a constituent unit of an oligonucleotide, means that a nucleobase is a sugar (for example, 2-deoxyribose, ribose, and a divalent organic group at the 2- and 4-position carbon atoms).
• nucleobase is not particularly limited as long as it is used for the synthesis of nucleic acids. and purine bases such as adenyl group (adenine base) and guanyl group (guanine base).
• optionally protected nucleobase means that, for example, an amino group, a carbonyl group, etc. in the nucleobase may be protected.
• the optionally protected nucleobase is preferably a nucleobase the amino group of which may be protected with the aforementioned amino group-protecting group, more preferably a nucleobase having no amino group, or the aforementioned amino group. It is a nucleobase having an amino group protected with a group-protecting group.
• carbonyl groups such as 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, dimethylcarbamic acid chloride, diethylcarbamic acid chloride, ethylphenylcarbamic acid chloride, 1-pyrrolidinecarboxylic acid chloride, 4-morpholinecarboxylic acid chloride, diphenylcarbamic acid chloride, etc., to obtain a carbonyl Groups can be protected.
• nucleobase as used herein means that a nucleobase is a substituent (e.g., a halogen atom, an alkyl group, an aralkyl group, an alkoxy group, an acyl group, an alkoxyalkyl group, a hydroxyl group, an amino group, a monoalkylamino, a dialkylamino , carboxy, cyano, nitro, etc.) substituted modified nucleobases (e.g., 8-bromoadenyl, 8-bromoguanyl, 5-bromocytosyl, 5-iodocytosyl, 5-bromouracil, 5-iodo uracil group, 5-fluorouracil group, 5-methylcytosyl group, 8-oxoguanyl group, hypoxanthinyl group, etc.) are also included.
• a substituent e.g., a halogen atom, an alkyl group, an aralkyl
• “Sugar” in the present specification also includes amino sugar in which the hydroxyl group is replaced by an amino group, and ribose in which the 2-position hydroxyl group is replaced by a halogen atom.
• amino sugars include, for example, 2-deoxyribose in which the 3-position hydroxyl group is replaced by an amino group, ribose in which the 3-position hydroxyl group is replaced by an amino group, and 3-position hydroxyl group is replaced by an amino group and the 2-position hydroxyl group is replaced by an amino group.
• amino sugars include, for example, 2-deoxyribose in which the 3-position hydroxyl group is replaced by an amino group, ribose in which the 3-position hydroxyl group is replaced by an amino group, and 3-position hydroxyl group is replaced by an amino group and the 2-position hydroxyl group is replaced by an amino group.
• examples thereof include ribose substituted with halogen (in the following formula, Xs represents a halogen atom).
• 2-deoxyribose or ribose in which the 2- and 4-position carbon atoms are bonded by a divalent organic group 2-deoxyribose or ribose in which the 3- and 5-position carbon atoms are bonded by a divalent organic group
• Examples of ribose, or 2-deoxyribose or ribose in which the 3- and 4-position carbon atoms are bonded by a divalent organic group include the following compounds.
• R represents a hydrogen atom, an optionally substituted hydrocarbon group, an optionally substituted hydroxyl group, or an optionally substituted amino group
• R′ represents a hydrogen atom or a hydroxyl group
• the hydroxyl group ( -OH ) in the phosphate group is replaced with a group (e.g., a protected phosphorus acid group) are also included in the "phosphate group".
• phosphite ester bond refers to a bond represented by the following formula (P1-1), or represented by the following formula (P1-2) in which the oxygen atom in the bond is replaced with NH. (In the formula below, * indicates the bonding position).
• phosphonate ester bond refers to a bond represented by the following formula (P2-1), or represented by the following formula (P2-2) in which the oxygen atom in the bond is replaced with NH. represents a bond (in the following formula, * indicates a bond position).
• phosphate ester bond refers to a bond represented by the following formula (P3-1), or represented by the following formula (P3-2) in which the oxygen atom in the bond is replaced with NH. represents a bond (in the following formula, * indicates a bond position).
• thiophosphate bond refers to a bond represented by the following formula (P4-1), or represented by the following formula (P4-2) in which the oxygen atom in the bond is replaced with NH. represents a bond (in the following formula, * indicates a bond position).
• nucleotide means a compound in which a phosphate group is bound to a nucleoside.
• the description of the phosphate group is as described above.
• oligonucleotide means a compound in which one or more nucleotides are linked to a nucleoside.
• the "oligonucleotide” includes not only oligonucleotides having a phosphate ester bond represented by the formula (P3-1), but also oligonucleotides having a phosphate ester bond represented by the formula (P3-2). Nucleotides, oligonucleosides having a thiophosphate ester bond represented by the above formula (P4-1), and oligonucleosides having a thiophosphate ester bond represented by the above formula (P4-2) are also included.
• the number of nucleosides in the oligonucleotide in the present invention is not particularly limited, it is preferably 2-50, more preferably 2-30.
• phosphoramidite refers to a phosphite diester monoamide (P(OR) 2 (NR 2 ), and each of the four R's independently represents an optionally substituted alkyl group. , said NR 2 may be combined with each other to form a cyclic amino group).
• phosphoramidated nucleoside refers to a group represented by —X n —P(OR)(NR 2 ) in a nucleoside (wherein X n represents an oxygen atom or NH , the three Rs each independently represent an optionally substituted alkyl group, and the NR 2 may combine with each other to form a cyclic amino group).
• X n represents an oxygen atom or NH
• the three Rs each independently represent an optionally substituted alkyl group, and the NR 2 may combine with each other to form a cyclic amino group.
• the term “phosphoramidated nucleotide” refers to a group represented by —X n —P (OR) (NR 2 ) in a nucleotide (wherein X n represents an oxygen atom or NH , the three Rs each independently represent an optionally substituted alkyl group, and the NR 2 may combine with each other to form a cyclic amino group).
• X n represents an oxygen atom or NH
• the three Rs each independently represent an optionally substituted alkyl group, and the NR 2 may combine with each other to form a cyclic amino group.
• X n represents an oxygen atom or NH
• the three Rs each independently represent an optionally substituted alkyl group
• the NR 2 may combine with each other to form a cyclic amino group.
• phosphoramidated oligonucleotide refers to a group represented by —X n —P(OR)(NR 2 ) in an oligonucleotide (wherein X n is an oxygen atom or NH , and each of the three Rs independently represents an optionally substituted alkyl group, and the NR 2 is two Rs that are bonded to each other to form a cyclic amino group).
• X n is an oxygen atom or NH
• each of the three Rs independently represents an optionally substituted alkyl group
• the NR 2 is two Rs that are bonded to each other to form a cyclic amino group
• oligonucleotide precursor having a phosphite ester bond means a precursor in which the phosphate ester bond in the oligonucleotide is replaced with a phosphite ester bond. Oligonucleotides with phosphate ester linkages can be produced by oxidizing this precursor.
• Oligonucleotide precursors with phosphite ester linkages are preferably phosphorous acid, obtained by condensation of (1) nucleosides, nucleotides or oligonucleotides with phosphoramidized nucleosides, nucleotides or oligonucleotides or (2) an oligonucleotide precursor having a phosphite ester bond obtained by condensing an oligonucleotide with a phosphoramidite.
• Oligonucleotide precursors with phosphite ester linkages are more preferably obtained by condensation of nucleosides, nucleotides or oligonucleotides with phosphoramidated nucleosides, nucleotides or oligonucleotides, phosphite ester linkages is an oligonucleotide precursor having
• Oligonucleotide precursors having a phosphite ester bond are more preferably protected with a nucleoside, nucleotide or oligonucleotide (a) having a hydrophobic protecting group and a temporary protecting group in which the hydroxyl group is removable under acidic conditions.
• nucleosides, nucleotides or oligonucleotide precursors with phosphite ester linkages obtained by condensation with nucleosides, nucleotides or oligonucleotides (b) which are polymorphic and phosphoramidized.
• the condensation includes condensation in which the oligonucleotide chain is extended in the direction from the 3′ end to the 5′ end (hereinafter referred to as “3′-5′ condensation”) and oligonucleotide chain extension in the direction from the 5′ end to the 3′ end.
• Any condensation hereinafter referred to as “5′-3′ condensation” that extends the nucleotide chain may be used.
• Preferred nucleosides, nucleotides or oligonucleotides (a) and phosphoramidized nucleosides, nucleotides or oligonucleotides (b) in each of 3′-5′ condensation and 5′-3′ condensation are described below in order. do.
• nucleoside, nucleotide or oligonucleotide (a) is, for example, the formula (a-I):
• r represents an integer of 0 or more; r + 1 Base 1 each independently represents a nucleobase that may be protected; r+1 X n1 each independently represent an oxygen atom or NH; r+1 X n2 each independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group that binds to the 2- and 4-position carbon atoms; r R 10 each independently represents an oxygen atom or a sulfur atom; r R p1 each independently represents a phosphate-protecting group; Pg is **L-Y-Z (wherein ** represents the bonding position with X n1 ), **L-Y-Sp (wherein ** represents the bonding position with X n1 , and Sp indicates a solid phase support) or a solid phase support; is. ] and compounds represented by (ie, nucleosides or oligonucleotides).
• compound (aI) is a nucleoside, and when r is 1 or more, compound (aI) is an oligonucleotide.
• r is preferably 49 or less, more preferably 29 or less, still more preferably 19 or less, particularly preferably 4 or less, and most preferably 2 or less.
• the amino group of Base 1 which is a nucleobase, is preferably protected with a protecting group.
• the amino group-protecting group include the aforementioned amino group-protecting groups, and the amino group-protecting group of Base 1 is preferably an acetyl group, a phenoxyacetyl group, a 4-isopropylphenoxyacetyl group, a benzoyl group, isobutyryl group, (2-hexyl)decanoyl group, dimethylformamidinyl group, and ⁇ NC(R 11 )—N(R 12 )(R 13 ) (wherein R 11 represents a methyl group; , R 12 and R 13 each independently represent a C 1-6 alkyl group, or R 11 and R 12 together represent a 5- or 6-membered nitrogen-containing group together with the carbon and nitrogen atoms to which they are attached. may form a heterocyclic ring).
• compound (aI) has a plurality of amino groups, the number of amino-protecting
• the halogen atom of Xn2 is preferably a fluorine atom or a chlorine atom, more preferably a fluorine atom.
• the optionally protected hydroxyl-protecting group of X n2 includes, for example, the hydroxyl-protecting groups described above, preferably a methyl group, a 2-methoxyethyl group, a triethylsilyl group, a triisopropylsilyl group, or a tert -butyldimethylsilyl group, (2-cyanoethoxy)methyl group and 1-(2-cyanoethoxy)ethyl group.
• the "divalent organic group bonded to the 2-position carbon atom and the 4-position carbon atom" of Xn2 is not particularly limited as long as it bonds to the 2-position carbon atom and the 4-position carbon atom of the nucleoside.
• the divalent organic group include an optionally substituted C 2-7 alkylene group, —O—, —NR 33 — (R 33 represents a hydrogen atom or a C 1-6 alkyl group), -S-, -CO-, -COO-, -OCONR 34 - (R 34 represents a hydrogen atom or a C 1-6 alkyl group) and -CONR 35 - (R 35 represents a hydrogen atom or a C 1-6 alkyl group and a divalent organic group composed of an optionally substituted C 1-7 alkylene group, and the like.
• the “divalent organic group bonded to the 2-position carbon atom and the 4-position carbon atom” includes an optionally substituted C 2-7 alkylene group, —OR i — (R i is bonded to the 4-position carbon atom a C 1-6 alkylene group), -O-NR 33 -R j - (R j represents a C 1-6 alkylene group bonded to the 4-position carbon atom, and R 33 has the same meaning as defined above), - OR k -OR l - (R k represents a C 1-6 alkylene group and R l represents a C 1-6 alkylene group that bonds and bridges with the 4-position carbon atom) is preferred, and -OR i - (R i has the same meaning as above), —O—NR 33 —R j — (R j and R 33 have the same meaning as above), —OR k —OR l — (R k and R l has the same meaning as above) is more preferred.
• the “divalent organic group bonded to the 2- and 4-position carbon atoms” includes —O—CH 2 —, —O—CH 2 —CH 2 —, —O—NR 33 —CH 2 —(R 33 has the same meaning as above), -O-CH 2 -O-CH 2 - is more preferred, -O-CH 2 -, -O-CH 2 -CH 2 -, -O-NH-CH 2 -, —O—N(CH 3 )—CH 2 — and —O—CH 2 —O—CH 2 — (each of which the left side binds to the 2-position carbon atom and the right side binds to the 4-position carbon atom) are further preferable.
• Each of r+1 X n2 is independently preferably a hydrogen atom, a halogen atom, or an optionally protected hydroxyl group, more preferably a hydrogen atom, a fluorine atom, or an optionally protected hydroxyl group.
• Examples of the r number of R p1 include the aforementioned phosphoric acid group-protecting group, preferably a 2-cyanoethyl group.
• compound (aI) is a nucleoside or oligonucleotide supported on a solid support.
• Pg is preferably **LYZ (in the formula, ** indicates the binding position with Xn1 ) or a solid phase carrier, more preferably **LYZ.
• the definitions and explanations of L, Y and Z are the same as the definitions and explanations of L, Y and Z for protecting group (Pg-5) above.
• the phosphoroamidated nucleoside, nucleotide or oligonucleotide (b) is, for example, the formula (bI):
• s represents an integer of 0 or more; s + 1 Base 2 each independently represents a nucleic acid base that may be protected; s+1 X n3 each independently represent an oxygen atom or NH; s+1 X n4 each independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group that binds to the 2- and 4-position carbon atoms; Q′′ represents a temporary protecting group removable under acidic conditions for the hydroxyl group; s R 14 each independently represents an oxygen atom or a sulfur atom; s+1 R p2 each independently represent a phosphate-protecting group; and R 15 and R 16 each independently represent an alkyl group or form together with an adjacent nitrogen atom It represents a 5- or 6-membered saturated cyclic amino group, and such a saturated cyclic amino group may have one oxygen atom or one sulfur atom as a ring-constituting atom in addition to the nitrogen atom.
• Base 2 each independently represents
• compound (bI) When s is 0, compound (bI) is a phosphoramidated nucleoside, and when s is 1 or more, compound (bI) is a phosphoramidite oligonucleotide. be.
• s is preferably 49 or less, more preferably 29 or less, still more preferably 19 or less, particularly preferably 4 or less, and most preferably 2 or less.
• the amino group of Base 2 which is a nucleobase, is preferably protected with a protecting group.
• the protecting group the -LYZ group described above can be used in addition to the protecting group for the amino group described above.
• compound (bI) has a plurality of
• Xn4 in the formula (bI) is the same as the description of Xn2 in the formula (aI).
• Each of s+1 X n4 is independently preferably a hydrogen atom, a halogen atom, or an optionally protected hydroxyl group, more preferably a hydrogen atom or an optionally protected hydroxyl group.
• R p2 in formula (b-I) above is the same as the description of R p1 in formula (a-I) above.
• Examples of s+1 R p2 include the aforementioned phosphate group-protecting groups, preferably a 2-cyanoethyl group.
• R 15 and R 16 in the above formula (bI) are each independently preferably a C 1-10 alkyl group or a 5- or 6-membered saturated group formed together with adjacent nitrogen atoms. It is a cyclic amino group, more preferably a C 1-10 alkyl group, still more preferably a C 1-6 alkyl group.
• Compound (aI) and compound (bI) can be produced by a known method (eg, the method described in International Publication No. 2017/104836) or a method analogous thereto.
• An oligonucleotide precursor having a phosphite ester bond obtained by condensation of compound (aI) and compound (bI) is a compound represented by the following formula (cI) (the following formula The definitions and explanations for the symbols (cI) are as described above).
• compound (cI) is an oligonucleotide precursor supported on a solid support.
• Pg is preferably **LYZ (in the formula, ** indicates the binding position with Xn1 ) or a solid phase carrier, more preferably **LYZ.
• At least one of s R 14 and r R 10 is preferably a sulfur atom.
• nucleoside, nucleotide or oligonucleotide (a) is, for example, the formula (a-I'):
• r represents an integer of 0 or more; r + 1 Base 1 each independently represents a nucleobase that may be protected; r X n1 each independently represents an oxygen atom or NH; r+1 X n2 each independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group that binds to the 2- and 4-position carbon atoms; X n5 represents a hydroxyl group or an amino group; r R 10 each independently represents an oxygen atom or a sulfur atom; r R p1 each independently represents a phosphate-protecting group; Pg is **L-Y-Z (in the formula, ** indicates the bonding position with the oxygen atom), **L-Y-Sp (wherein, ** indicates the bonding position with X n1 , and Sp indicates a solid phase support) or a solid phase support; are the same. ] and compounds represented by (ie, nucleosides or oligonu).
• Xn5 in formula (aI') is preferably a hydroxyl group.
• the explanation of the symbols other than X n5 is the same as the explanation of the symbols in the above formula (aI).
• Pg in the formula (aI') is **LY-Sp (wherein ** represents the binding position with X n1 and Sp represents the solid phase carrier) or a solid phase carrier
• compound (aI') is a nucleoside or oligonucleotide supported on a solid support.
• Pg is preferably **LYZ (in the formula, ** indicates the bonding position with an oxygen atom) or a solid phase carrier, more preferably **LYZ.
• nucleoside, nucleotide or oligonucleotide (b) is, for example, the formula (b-I'):
• s represents an integer of 0 or more; s + 1 Base 2 each independently represents a nucleic acid base that may be protected; s X n3 each independently represent an oxygen atom or NH; s+1 X n4 each independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group that binds to the 2- and 4-position carbon atoms; X n6 represents a hydroxyl group protected with a temporary protecting group removable under acidic conditions, or an amino group protected with a temporary protecting group removable under acidic conditions; s R 14 each independently represents an oxygen atom or a sulfur atom; s+1 R p2 each independently represent a phosphate protecting group; and R 15 and R 16 each independently represent an alkyl group or form together with an adjacent nitrogen atom It represents a 5- or 6-membered saturated cyclic amino group, and such a saturated cyclic amino group may have one oxygen atom or one sulfur atom as a ring
• Xn6 is preferably a hydroxyl group protected with a temporary protecting group removable under acidic conditions.
• the explanation of the symbols other than Xn6 is the same as the explanation of the symbols in the above formula (bI).
• Compound (a-I') and compound (b-I') can be produced by a known method (eg, the method described in International Publication No. 2017/104836) or a method analogous thereto.
• An oligonucleotide precursor having a phosphite ester bond obtained by condensation of compound (a-I') and compound (b-I') has the following formula (c-I'):
• r represents an integer of 0 or more; r + 1 Base 1 each independently represents a nucleobase that may be protected; r+1 X n1 each independently represent an oxygen atom or NH; r+1 X n2 each independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group that binds to the 2- and 4-position carbon atoms; r R 10 each independently represents an oxygen atom or a sulfur atom; r R p1 each independently represents a phosphate-protecting group; s represents an integer of 0 or more; s + 1 Base 2 each independently represents a nucleic acid base that may be protected; s X n3 each independently represent an oxygen atom or NH; s+1 X n4 each independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group that binds to the 2- and 4-position carbon atoms;
• Pg in the formula (cI′) is **LY-Sp (wherein ** indicates the binding position with X n1 and Sp indicates a solid phase carrier) or a solid phase carrier
• compound (cI') is an oligonucleotide precursor supported on a solid support.
• Pg is preferably **LYZ (in the formula, ** indicates the bonding position with an oxygen atom) or a solid phase carrier, more preferably **LYZ.
• At least one of r R 10 and s R 14 is preferably a sulfur atom.
• an oligonucleotide precursor having a phosphite ester bond obtained by condensing an oligonucleotide with a phosphoramidite will be explained.
• the precursor include formula ( ⁇ -I) or formula ( ⁇ -I′):
• t represents an integer of 1 or more, t + 1
• Base 1 each independently represents a nucleobase that may be protected
• t+1 X n1 each independently represents an oxygen atom or NH
• t+1 X n2 each independently represents a hydrogen atom, a halogen atom, an optionally protected hydroxyl group, or a divalent organic group that binds to the 2- and 4-position carbon atoms
• t R 10 each independently represents an oxygen atom or a sulfur atom
• t R p1 and R p3 each independently represent a phosphate group-protecting group
• R p4 represents an alkyl group having a protected amino group, or a protecting group for a phosphate group
• Pg is **L-Y-Z (in the formula, ** indicates the bonding position with X n1 or the bonding position with the oxygen atom), **L-Y-Sp (wherein ** is X n1 , and Sp indicates a solid phase support) or a
• Pg in the formula ( ⁇ -I) or formula ( ⁇ -I′) is **LY-Sp (wherein ** represents the binding position with Xn1 , and Sp represents the solid phase carrier ) or a solid phase support, compound ( ⁇ -I) or compound ( ⁇ -I′) is an oligonucleotide precursor supported on the solid phase support.
• Pg is preferably **LYZ (in the formula, ** indicates the bonding position with an oxygen atom) or a solid phase carrier, more preferably **LYZ.
• t is preferably 49 or less, more preferably 29 or less, still more preferably 19 or less, particularly preferably 4 or less, and most preferably 2 or less.
• Examples of the "phosphate group-protecting group" of R p3 and R p4 include the aforementioned phosphate group-protecting groups.
• R p3 is preferably a 2-cyanoethyl group.
• the "protected amino group” in the "alkyl group having a protected amino group” of R p4 includes, for example, an amino group protected by the aforementioned amino group-protecting group.
• the “alkyl group having a protected amino group” for R p4 is preferably a C 1-6 alkyl group having a protected amino group, more preferably a 6-(trifluoroacetylamino)hexyl group.
• R p4 is preferably a 2-cyanoethyl group or a 6-(trifluoroacetylamino)hexyl group. Descriptions of other symbols are as described above.
• oligonucleotide precursor having a phosphonate ester bond means a precursor in which the phosphate ester bond in the oligonucleotide is replaced with a phosphonate ester bond. Oligonucleotides with phosphate ester linkages can be produced by oxidizing this precursor.
• Oligonucleotide precursors with phosphonate ester linkages are preferably nucleosides, nucleotides or oligonucleotides (a) with hydrophobic protecting groups and H - oligonucleotide precursors with phosphonate ester linkages obtained by condensation with phosphonates (B).
• the H-phosphonate (B) may be used in the form of a salt such as triethylamine salt.
• the condensation may be either 3'-5' condensation or 5'-3' condensation.
• Preferred nucleosides, nucleotides or oligonucleotides (a) and H-phosphonates (B) for 3'-5' condensation and 5'-3' condensation, respectively, are described in order.
• nucleoside, nucleotide or oligonucleotide (a) is preferably compound (aI) as described above.
• the H-phosphonate (B) includes, for example, compounds represented by the following formula (BI) (definition and explanation of symbols in the following formula (BI) are , the same as the definition and explanation of the symbols in the formula (bI).).
• Compound (BI) can be produced by a known method (eg, the method described in Acta Biochimica Polonica. 1998, 45, 907-915.) or a method analogous thereto.
• the oligonucleotide precursor having a phosphonate ester bond obtained by condensation of the compound (aI) and the compound (BI) is a compound represented by the following formula (C-I) (the following formula (C -I) is the same as the definition and explanation of the symbols in formula (cI) above).
• compound (C-I) is an oligonucleotide precursor supported on a solid support.
• Pg is preferably **LYZ (in the formula, ** indicates the binding position with Xn1 ) or a solid phase carrier, more preferably **LYZ.
• At least one of s R 14 and r R 10 is preferably a sulfur atom.
• nucleoside, nucleotide or oligonucleotide (a) is preferably the aforementioned compound (a-I').
• the H-phosphonate (B) includes, for example, compounds represented by the following formula (BI′) (definition and explanation of the symbols of the following formula (BI′) are , the same as the definitions and explanations of the symbols in the above formula (bI′)).
• Compound (BI') is produced by a known method (for example, the method described in Phosphorus Chemistry II. Topics in Current Chemistry, vol 361. Springer, Cham. Recent Advances in H-Phosphonate Chemistry) or a method analogous thereto. be able to.
• An oligonucleotide precursor having a phosphonate ester bond obtained by condensation of compound (aI') and compound (BI') is a compound represented by the following formula (CI') (
• the definitions and explanations of the symbols in the formula (C-I') are the same as the definitions and explanations of the symbols in the formula (c-I').).
• Pg in the formula (C-I') is **LY-Sp (wherein ** represents the binding position with Xn1 and Sp represents a solid phase carrier) or a solid phase carrier
• compound (CI) is an oligonucleotide precursor supported on a solid support.
• Pg is preferably **LYZ (in the formula, ** indicates the bonding position with an oxygen atom) or a solid phase carrier, more preferably **LYZ.
• At least one of r R 10 and s R 14 is preferably a sulfur atom.
• one-pot synthesis refers to a synthesis consisting of multiple steps, including a step of synthesizing an intermediate and a step of synthesizing the desired final product, wherein the synthesis does not isolate said intermediate. means.
• the "step of synthesizing an intermediate” may be one or two or more.
• a compound represented by (excluding 2,2-dipyridyl disulfide) to oxidize an oligonucleotide precursor having a phosphite ester bond or a phosphonate ester bond to obtain an oligo having a phosphate ester bond It is characterized by producing nucleotides.
• Compound (I) may be used alone or in combination of two or more.
• Oxidizing Agent Compound (I) (excluding 2,2-dipyridyl disulfide), which is an oxidizing agent used in the production method of the present invention, will be described below.
• R 1 in the formula (I) is a halogen atom
• R 2 in the formula (I) is an optionally substituted aryl group or a substituted is a good heteroaryl group.
• R 1 in the formula (I) is an optionally substituted alkyl group, an optionally substituted aryl group, or a substituted is an optionally substituted heteroaryl group
• R 2 in the above formula (I) is an optionally substituted aryl group or an optionally substituted heteroaryl group.
• R 1 and R 2 in the above formula (I) form an optionally substituted heterocyclic ring together with the sulfur atom and oxygen atom to which they are attached.
• R 1 in formula (I) is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group; ), together with the sulfur atom and nitrogen atom to which they are attached, form an optionally substituted heterocyclic ring, or ( 2 ) R 1 and R 3 in the formula ( I) together with the nitrogen atom to which they are attached form an optionally substituted heterocyclic ring, and R 2 in the above formula (I) is an optionally substituted aryl group or an optionally substituted hetero It is an aryl group.
• X 1 is —N(—R 3 )—, and R 1 is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted a heteroaryl group, and R 2 and R 3 form an optionally substituted heterocyclic ring together with the sulfur atom and nitrogen atom to which they are attached (herein referred to as "compound (IA)") described), and in the above formula (I), X 1 is a sulfur atom, and R 1 is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, and R 2 is an optionally substituted aryl group or an optionally substituted heteroaryl group (excluding 2,2-dipyridyl disulfide) (hereinafter referred to as compounds ( IB)”) is preferred.
• X 1 is a sulfur atom or -N(-R 3 )-, R 1 is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group;
• R 2 is an optionally substituted aryl group or an optionally substituted heteroaryl group, and when X 1 is -N(-R 3 )-, R 2 and R 3 are preferably compounds (excluding 2,2-dipyridyl disulfide) that form an optionally substituted heterocyclic ring together with the sulfur atom and nitrogen atom to which they are attached.
• compound (IA) and compound (IB) will be described below.
• R 1 in formula (I) is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, preferably an optionally substituted C 1-10 alkyl group, an optionally substituted phenyl group, an optionally substituted 5- or 6-membered monocyclic heteroaryl group, or an optionally substituted 8-14 is a membered fused polycyclic heteroaryl group.
• R 2 and R 3 in formula (I) form an optionally substituted heterocyclic ring together with the sulfur atom and nitrogen atom to which they are attached.
• 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.
• R 1a in the formula (Ia) is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, preferably optionally substituted It is a C 1-10 alkyl group, an optionally substituted phenyl group, or an optionally substituted 5- or 6-membered monocyclic heteroaryl group.
• Ring A in the formula (Ia) is a 5- or 6-membered unsaturated heterocyclic ring.
• n in the above formula (Ia) is an integer of 0 to 3
• m R 2a are each independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron It is an withdrawing group, preferably an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, or an electron withdrawing group.
• "m is 0" means that R2a is absent.
• the 5- or 6-membered unsaturated heterocyclic ring for ring A is a ring represented by any one of the following formulas (r2) to (r4) [the symbols in the formulas are defined as described above. ]. Note that R 2a is omitted in the following formulas (r2) to (r4).
• the condensed ring formed by two adjacent R 2a and ring A is preferably a bicyclic or tricyclic condensed ring, more preferably a bicyclic condensed ring.
• Examples of the condensed ring include rings represented by any one of the following formulas (r5) to (r25).
• the fused ring may be substituted.
• Compound (Ia) is preferably a compound represented by the following formula (Ic).
• R 1c in the formula (Ic) is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, preferably optionally substituted It is a C 1-6 alkyl group, an optionally substituted phenyl group, or an optionally substituted 5- or 6-membered monocyclic heteroaryl group.
• R 1c is preferably selected from the group consisting of (1) an optionally substituted phenyl group and an optionally substituted 5- or 6-membered monocyclic heteroaryl group; a C 1-6 alkyl group optionally substituted with 1 or 2 substituents, (2) a phenyl 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 (3) 5- or 6-membered 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 a monocyclic heteroaryl group.
• R 1c is preferably (1) a group consisting of an optionally substituted phenyl group and an optionally substituted 5- or 6-membered monocyclic heteroaryl group a C 1-6 alkyl group optionally substituted with 1 or 2 substituents selected from (2) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; or (3) 1 to 1 selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group It is a pyridyl group optionally substituted with 4 substituents.
• R 1c is preferably (1) a C 1-6 alkyl group optionally substituted with 1 or 2 pyridyl groups, (2) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom and a nitro group; or (3) an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom, and a nitro group. It is a pyridyl group optionally substituted with 1 to 4 substituents.
• R 1c is preferably (1) a C 1-6 alkyl group optionally substituted with 1 or 2 pyridyl groups, (2) a phenyl group optionally substituted with 1 or 2 C 1-6 alkyl groups, or (3) a pyridyl group (especially a 2-pyridyl group or a 4-pyridyl group) is.
• R 1c is preferably (1) a C 1-6 alkyl group, (2) a phenyl group optionally substituted with 1 or 2 substituents selected from the group consisting of a C 1-6 alkyl group, a C 1-6 alkoxy group and a halogen atom, or (3) 1 C A pyridyl group substituted with a 1-6 alkyl group (eg, 4-methyl-2-pyridyl group) is.
• p in the above formula (Ic) is 0 or 1, preferably 0;
• p is 0 means that —CH(R 2c )— in formula (Ic) does not exist.
• R 2c in the formula (Ic) is a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group, preferably a hydrogen atom or a substituted may be a C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, or an electron-withdrawing group.
• Ring C in the formula (Ic) is a 6-membered aromatic hydrocarbon ring, a 6-membered nitrogen-containing aromatic heterocycle, or a 10-membered bicyclic condensed aromatic heterocycle containing a nitrogen atom.
• the ring C is preferably a 6-membered aromatic hydrocarbon ring (i.e., benzene ring) or a 6-membered nitrogen-containing aromatic heterocyclic ring, more preferably a benzene ring or It is a pyridine ring, more preferably a pyridine ring.
• the ring C is preferably a benzene ring, a pyridine ring or a quinoline ring.
• q in the formula (Ic) is an integer of 0 to 4, preferably an integer of 0 to 2, more preferably 0 or 1.
• q is 0
• R3c is absent.
• each of the q R 3cs in the formula (Ic) are each independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group.
• each of the q R 3cs is preferably an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, or It is an electron-withdrawing group, more preferably a halogen atom or a nitro group, still more preferably a nitro group.
• q R 3c are each independently preferably an electron-withdrawing group, more preferably a halogen atom, a C 1-6 perfluoroalkyl group or a nitro group , more preferably a C 1-6 perfluoroalkyl group or a nitro group, particularly preferably a trifluoromethyl group.
• Suitable compounds (Ic) include the following compounds. Each of the following compounds may be used alone or in combination of two or more.
• R 1c is (1) optionally substituted with 1 or 2 substituents selected from the group consisting of an optionally substituted phenyl group and an optionally substituted 5- or 6-membered monocyclic heteroaryl group; a good C 1-6 alkyl group, (2) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; or (3) 1 to 1 selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group
• R 1c is (1) a C 1-6 alkyl group optionally substituted with 1 or 2 pyridyl groups, (2) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom and a nitro group; or (3) an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom, and a nitro group.
• a pyridyl group optionally substituted with 1 to 4 substituents
• p is 0,
• Ring C is a benzene ring or a pyridine ring
• q is an integer from 0 to 2
• a compound in which q R 3c are each independently a halogen atom or a nitro group (referred to herein as “compound (Ic-iii)”).
• R 1c is (1) optionally substituted with 1 or 2 substituents selected from the group consisting of an optionally substituted phenyl group and an optionally substituted 5- or 6-membered monocyclic heteroaryl group; a good C 1-6 alkyl group, (2) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; or (3) 1 to 1 selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group
• R 1c is (1) a C 1-6 alkyl group, (2) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom and a nitro group; or (3) an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom, and a nitro group.
• a pyridyl group optionally substituted with 1 to 4 substituents
• p is 0 or 1
• Ring C is a benzene ring, a pyridine ring or a quinoline ring
• q is an integer from 0 to 2
• a compound in which q R 3c are each independently a halogen atom, a C 1-6 perfluoroalkyl group or a nitro group referred to herein as “compound (Ic-iii′)”).
• Preferable specific examples of the compound (Ic) include compounds represented by any one of the following formulas (Ic-1) to (Ic-17).
• compound (Ic) is preferably 2-phenylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-1)), 2-isopropyl isothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-2)), 2-isopropylisothiazolo[4,5-c]pyridin-3(2H)-one (compound (Ic -3)), 2-(2-pyridyl)-1,2-benzothiazol-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 (compound (Ic-6) ), 5-
• compound (Ic) is preferably 2-phenylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-1)), 2 - isopropylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-2)), 2-isopropylisothiazolo[4,5-c]pyridin-3(2H)-one (compound (Ic-3)), 2-(2-pyridyl)-1,2-benzothiazol-3(2H)-one (compound (Ic-4)), 5-nitro-2-phenyl-1,2-benzo Thiazol-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 (compound (Ic-6)), 5-nitro
• compound (Ic) is preferably 2-phenylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-1)), 2 -isopropylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-2)), 5-nitro-2-phenyl-1,2-benzothiazol-3(2H)-one ( Compound (Ic-5)), 2-phenylisothiazolo[4,5-c]pyridin-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]pyridin-3(2H)-one (compound (Ic-12)) , 2-phenyl-2H-1,2-benzothiazin-3(4
• compound (Ic) is preferably 2-phenylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-1)), 2 - isopropylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-2)), 2-phenylisothiazolo[4,5-c]pyridin-3(2H)-one (compound (Ic-10)), 2-phenylisothiazolo[5,4-b]quinolin-3(2H)-one (compound (Ic-11)), 2-phenyl-5-trifluoromethylisothiazolo[5, 4-b]pyridin-3(2H)-one (compound (Ic-12)), 2-phenyl-2H-1,2-benzothiazin-3(4H)-one (compound (Ic-13)), 2- (2,6-dimethylphenyl)isothiazolo[5,4
• compound (Ic) is most preferably 2-phenylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-1)).
• R 1 in formula (I) is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, preferably an optionally substituted C 1-10 alkyl group, an optionally substituted phenyl group, an optionally substituted 5- or 6-membered monocyclic heteroaryl group, or an optionally substituted 8-14 is a membered fused polycyclic heteroaryl group.
• R 2 in formula (I) above is an optionally substituted aryl group or an optionally substituted heteroaryl group, preferably an optionally substituted phenyl group, substituted is an optionally substituted 5- or 6-membered monocyclic heteroaryl group, or an optionally substituted 8- to 14-membered condensed polycyclic heteroaryl group.
• Compound (Ib) is preferably a compound represented by formula (Ib) below (excluding 2,2-dipyridyl disulfide). Compound (Ib) may be used alone or in combination of two or more.
• R 1b in the formula (Ib) is an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group, preferably optionally substituted C 1-10 alkyl group, optionally substituted phenyl group, optionally substituted 5- or 6-membered monocyclic heteroaryl group, or optionally substituted 8- to 14-membered condensed polycyclic group It is a heteroaryl group.
• R 1b is more preferably (1) an alkyl group substituted with one carboxy group, (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; , (3) a 2-benzimidazolyl group optionally substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group and an electron-withdrawing group; , (4) 2-benzoxoxa 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; solyl group, (5) optionally substituted alkyl group, optionally substituted alkoxy group and 5-(1 , 2,3-triazoly
• R 1b is more preferably (1) an alkyl group substituted with one carboxy group, (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; , (3) a 2-benzimidazolyl group optionally substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group and an electron-withdrawing group; , (4) 2-benzoxoxa 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; zolyl group, (5) optionally substituted alkyl group, optionally substituted alkoxy group and 5-(1 , 2,3-triazolyl) group, (6) a 1H-tetrazol-5-yl group optional
• R 1b is particularly preferably (1) a C 1-6 alkyl group substituted with one carboxy group, (2) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzothiazolyl group, which may be (3) substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzimidazolyl group, which may be (4) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzoxazolyl group, which may be (5) substituted with 1 or 2 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6
• R 1b is most preferably C 1-6 alkyl group substituted with one carboxy group, 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 5- (1,2,3-triazolyl) group, 1-phenyl-1H-tetrazol-5-yl group, (pyridin-N-oxide)-2-yl group, 4-pyridyl group, or 2-pyridyl group.
• R 1b is more preferably (1) an alkyl group substituted with one carboxy group, (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; , (3) a 2-benzimidazolyl group optionally substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group and an electron-withdrawing group; , (4) 2-benzoxoxa 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; solyl group, (5) a 1H-tetrazol-5-yl group optionally substituted with one substituent selected from the group consisting of
• R 1b is more preferably (1) a C 1-6 alkyl group substituted with one carboxy group, (2) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzothiazolyl group, which may be (3) substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzimidazolyl group, which may be (4) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzoxazolyl group, which may be (5) a 1H-tetrazol-5-yl group optionally substituted with one substituent selected from the group consisting of an optionally substituted
• R 1b is particularly preferably C 1-6 alkyl group substituted with one carboxy group, 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 1- phenyl-1H-tetrazol-5-yl group, (pyridin-N-oxide)-2-yl group, 4-pyridyl group, or 2-pyridyl group.
• Ring B in the formula (Ib) is a 6-membered aromatic hydrocarbon ring (i.e., benzene ring) or a 5- or 6-membered aromatic heterocyclic ring, preferably a 5- or 6-membered aromatic heterocyclic ring , more preferably a triazole ring, a tetrazole ring, a pyridine ring or a pyridine-N-oxide ring, still more preferably a tetrazole ring, a pyridine ring or a pyridine-N-oxide ring.
• n in the formula (Ib) is an integer of 0 to 5, and each of the n R 2b is independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron It is an attracting group, more preferably an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, or an electron withdrawing group.
• n is 0
• R2b is absent.
• n in the formula (Ib) is an integer of 2 or more, two adjacent R 2b together with ring B are an optionally substituted bicyclic condensed aromatic heterocyclic ring (preferably substituted optionally substituted benzimidazole ring, optionally substituted benzoxazole ring, or optionally substituted benzothiazole ring).
• 2-benzothiazolyl group which may be (2) a 2-benzimidazolyl group optionally substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group and an electron-withdrawing group; , (3) 2-benzoxoxa 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; zolyl group, (4) optionally substituted alkyl group, optionally substituted alkoxy group and 5-(1 , 2,3-triazolyl) group, (5) a 1H-tetrazol-5-yl group optionally substituted with one substituent selected from the group consisting of an optionally substituted alkyl group and an optionally substituted
• the group (r1) is more preferably (1) 1 to 4 selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group and an electron-withdrawing group 2-benzothiazolyl group optionally substituted with one substituent, (2) a 2-benzimidazolyl group optionally substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group and an electron-withdrawing group; , (3) 2-benzoxoxa 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; zolyl group, (4) optionally substituted alkyl group, optionally substituted alkoxy group and 5-(1 , 2,3-triazolyl) group, (5) a 1H-tetrazol-5-yl group optionally substituted with one substituent selected from the group consisting of an optionally substituted alky
• the group (r1) is more preferably from (1) an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group a 2-benzothiazolyl group optionally substituted with 1 to 4 substituents selected from the group consisting of (2) substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzimidazolyl group, which may be (3) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzoxazolyl group, which may be (4) substituted with 1 or 2 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group
• 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-tetrazol-5-yl group, (pyridin-N-oxide)-2-yl group, 4-pyridyl group or 2-pyridyl group, most preferably 2-benzothiazolyl group, 2-benzimidazolyl group, 2- benzoxazolyl group, 5-(1,2,3-triazolyl) group, 1-phenyl-1H-tetrazol-5-yl group, or (pyridin-N-oxide)-2-yl group.
• the group (r1) is preferably (1) an optionally substituted alkyl group, optionally substituted a 2-benzothiazolyl group optionally substituted with 1 to 4 substituents selected from the group consisting of an alkoxy group and an electron-withdrawing group; (2) a 2-benzimidazolyl group optionally substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group and an electron-withdrawing group; , (3) 2-benzoxoxa 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; solyl group, (4) a 1H-tetrazol-5-yl group optionally substituted with one substituent selected from the group consisting of an optionally substituted alkyl group and an optionally substituted
• the group (r1) is more preferably selected from (1) an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group
• 2-benzothiazolyl group optionally substituted with 1 to 4 substituents selected from the group consisting of (2) substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group
• 2-benzimidazolyl group which may be (3) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group
• 2-benzoxazolyl group which may be (4) a 1H-tetrazol-5-yl group optionally substituted with one substituent selected from the group consisting of an optionally substituted C 1-6 alkyl group and an optionally substituted pheny
• the group (r1) is particularly preferably a 2-benzothiazolyl group, a 2-benzimidazolyl group, a 2-benzoxazolyl group, a 1-phenyl-1H-tetrazol-5-yl group, a (pyridine-N -oxid)-2-yl, 4-pyridyl, or 2-pyridyl, most preferably 2-benzothiazolyl, 2-benzimidazolyl, 2-benzoxazolyl, 1-phenyl-1H-tetrazole -5-yl group or (pyridin-N-oxide)-2-yl group.
• Suitable compounds (Ib) include the following compounds. Each of the following compounds may be used alone or in combination of two or more.
• R 1b is (1) an alkyl group substituted with one carboxy group, (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; , (3) a 2-benzimidazolyl group optionally substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group and an electron-withdrawing group; , (4) 2-benzoxoxa 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; zolyl group, (5) optionally substituted alkyl group, optionally substituted alkoxy group and 5-(1 , 2,3-triazolyl) group, (6) a 1H-t
• R 1b is (1) an alkyl group substituted with one carboxy group, (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; , (3) a 2-benzimidazolyl group optionally substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group and an electron-withdrawing group; , (4) 2-benzoxoxa 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; zolyl group, (5) optionally substituted alkyl group, optionally substituted alkoxy group and 5-(1 , 2,3-triazolyl) group, (6) a 1H-
• R 1b is (1) a C 1-6 alkyl group substituted with one carboxy group, (2) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzothiazolyl group, which may be (3) substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzimidazolyl group, which may be (4) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzoxazolyl group, which may be (5) substituted with 1 or 2 substituents selected from the group consisting of an optionally substituted C
• a 2-pyridyl group optionally substituted with 1 to 4 substituents and the group (r1) is (1) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group;
• 2-benzothiazolyl group which may be (2) substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group;
• 2-benzimidazolyl group which may be (3) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group;
• 2-benzoxazolyl group which may be (4) substituted with 1 or 2 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substitute
• R 1b is a C 1-6 alkyl group substituted with one carboxy group, 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 5-(1,2,3-triazolyl) group , 1-phenyl-1H-tetrazol-5-yl, (pyridin-N-oxide)-2-yl, 4-pyridyl, or 2-pyridyl, and group (r1) is 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 5-(1,2,3-triazolyl) group, 1-phenyl-1H-tetrazol-5-yl group, (pyridine-N-oxide)-2 -yl group, 4-pyridyl group, or 2-pyridyl group (referred to herein as “compound (Ib-iv)”).
• R 1b is a C 1-6 alkyl group substituted with one carboxy group, 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 5-(1,2,3-triazolyl) group , 1-phenyl-1H-tetrazol-5-yl, (pyridin-N-oxide)-2-yl, 4-pyridyl, or 2-pyridyl, and group (r1) is 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 5-(1,2,3-triazolyl) group, 1-phenyl-1H-tetrazol-5-yl group, or (pyridine-N-oxide)- It is a compound having a 2-yl group (sometimes referred to as “compound (Ib-v)” in this specification).
• R 1b is (1) an alkyl group substituted with one carboxy group, (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; , (3) a 2-benzimidazolyl group optionally substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted alkyl group, an optionally substituted alkoxy group and an electron-withdrawing group; , (4) 2-benzoxoxa 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; zolyl group, (5) a 1H-tetrazol-5-yl group optionally substituted with one substituent selected from the group consisting of an optionally substituted alkyl group and an
• R 1b is (1) a C 1-6 alkyl group substituted with one carboxy group, (2) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzothiazolyl group, which may be (3) substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzimidazolyl group, which may be (4) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group; 2-benzoxazolyl group, which may be (5) a 1H-tetrazol-5-yl group optionally substituted with one substituent
• a 2-pyridyl group optionally substituted with 1 to 4 substituents, and the group (r1) is (1) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group;
• 2-benzothiazolyl group which may be (2) substituted with 1 to 5 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group;
• 2-benzimidazolyl group which may be (3) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group and an electron-withdrawing group;
• 2-benzoxazolyl group which may be (4) a 1H-tetrazol-5-yl group optionally substituted with one substituent selected from the group consisting of an optionally
• R 1b is a C 1-6 alkyl group substituted with one carboxy group, 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 1-phenyl-1H-tetrazol-5-yl group , (pyridin-N-oxide)-2-yl group, 4-pyridyl group, or 2-pyridyl group, and group (r1) is 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl 1-phenyl-1H-tetrazol-5-yl group, (pyridin-N-oxide)-2-yl group, 4-pyridyl group, or 2-pyridyl group (herein referred to as “compound (Ib -iii')").
• R 1b is a C 1-6 alkyl group substituted with one carboxy group, 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 1-phenyl-1H-tetrazol-5-yl group , (pyridin-N-oxide)-2-yl group, 4-pyridyl group, or 2-pyridyl group, and group (r1) is 2-benzothiazolyl group, 2-benzimidazolyl group, 2-benzoxazolyl group, 1-phenyl-1H-tetrazol-5-yl group, or (pyridin-N-oxide)-2-yl group (herein referred to as "compound (Ib-iv')" There is).
• Preferred specific examples of the compound (Ib) include compounds represented by any of the following formulas (Ib-1) to (Ib-12) (Ph in the following formula (Ib-7) is a phenyl group; show).
• 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)), 5,5'-dithiobis (1 -phenyl-1H-tetrazole) (compound (Ib-7)), 2,2′-dithiobis(pyridine-N-oxide) (compound (Ib-8)), 4,4′-dipyridyl disulfide (com
• 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)), 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-1)), 2-(2-benzo
• 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′-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-1)), 2-(2-benz
• 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)), 5,5'-dithiobis At least one selected from the group consisting of (1-phenyl-1H-tetrazole) (compound (Ib-7)) and 2,2′-dithiobis(pyridine-N-oxide) (compound (Ib-8)) be.
• compound (Ib-1) 2,2′-di
• 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′-dithiobis(1-phenyl-1H-tetrazole) (compound (Ib-7)), and 2,2′-dithiobis It is at least one selected from the group consisting of (pyridine-N-oxide) (compound (Ib-8)).
• compound (Ib) is most preferably 2,2'-dibenzothiazolyl disulfide (compound (Ib-1)).
• Compound (ID), compound (IE), compound (IF) and compound (IG) may be used singly or in combination of two or more.
• Compound (ID), compound (IE), compound (IF) and compound (IG) are described in order below.
• Compound (ID) A compound (ID) in which X 1 in the above formula (I) is a single bond will be described.
• R 1d in the formula (Id) is preferably a chlorine atom.
• R 2d in the formula (Id) is preferably an optionally substituted heteroaryl group, more preferably an optionally substituted 5- or 6-membered monocyclic heteroaryl with an electron-withdrawing group. group, more preferably a pyridyl group optionally substituted with a nitro group.
• Suitable compounds (Id) include the following compounds. Each of the following compounds may be used alone or in combination of two or more.
• R 1d is a chlorine atom
• R 2d is a 5- or 6-membered monocyclic heteroaryl group optionally substituted with an electron-withdrawing group (this specification Described as “compound (Id-ii)” in the book).
• a preferred specific example of the compound (Id) is 3-nitro-2-pyridinesulfenyl chloride represented by the following formula (Id-1).
• R 1e and R 2e each independently represent an optionally substituted alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group.
• R 1e and R 2e in the formula (Ie) are each independently preferably an optionally substituted aryl group, more preferably an optionally substituted phenyl group.
• a preferred specific example of compound (Ie) is (S)-phenyl benzenethiosulfonate represented by the following formula (Ie-1).
• 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 R 1f each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group
• R 2f represents a hydrogen atom, an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group
• v represents an integer of 0-3.
• It is a compound represented by Compound (If) may be used alone or in combination of two or more.
• Ring D in the formula (If) is preferably a 6-membered aromatic hydrocarbon ring (that is, a benzene ring).
• w in the formula (If) is preferably an integer of 0 to 2, more preferably 0 or 1.
• "w is 0" means that R 1f does not exist.
• Each of w R 1f in the above formula (If) is preferably an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom or nitro is the base.
• R 2f in the formula (If) is preferably a hydrogen atom, an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, or an electron-withdrawing group , more preferably a hydrogen atom, an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom or a nitro group.
• v in the above formula (If) is preferably 0.
• “v is 0” means that —CH 2 — in parentheses in the above formula (If) does not exist.
• Suitable compounds include the following compounds. Each of the following compounds may be used alone or in combination of two or more.
• Ring D is a benzene ring
• w is preferably an integer of 0 to 2
• w R 1f are each independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group can be
• Ring D is a benzene ring
• w is preferably an integer of 0 to 2
• w R 1f are each independently an optionally substituted C 1-6 alkyl group or an optionally substituted C 1-6 alkoxy group , a halogen atom or a nitro group
• v is an integer from 0 to 3 (referred to herein as "compound (If-ii)").
• 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
• a preferred specific example of the compound (If) is 3H-2,1-benzoxathiol-3-one represented by the following formula (If-1).
• Ring E represents a 4- to 8-membered nitrogen-containing heterocyclic ring
• x represents an integer from 0 to 6
• x R 1g each independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, an oxo group, or an electron-withdrawing group
• x is an integer of 2 or more
• two adjacent R 1g together with ring E may form an optionally substituted bicyclic condensed nitrogen-containing heterocyclic ring
• R 2g is , 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.
• R 2g in the formula (Ig) is preferably an optionally substituted heteroaryl group, more preferably an optionally substituted 5- or 6-membered monocyclic nitrogen-containing heteroaryl group, or It is an optionally substituted 8- to 14-membered condensed polycyclic nitrogen-containing heteroaryl group.
• R 2g is more preferably (1) 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.
• optionally substituted 2-pyridyl group or (2) 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 It is a 2-benzothiazolyl group which may be substituted.
• R 2g is particularly preferably (1) 1 to 1 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 a 2-pyridyl group optionally substituted by 4 substituents, or (2) an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom, and a 2-benzothiazolyl group optionally substituted with 1 to 4 substituents selected from the group consisting of a nitro group.
• R 2g is most preferably a 2-pyridyl group or a 2-benzothiazolyl group.
• x in the formula (Ig) is an integer of 0-6, preferably an integer of 2-6.
• x is 0" means that R 1g is absent.
• x R 1g in the formula (Ig) are each independently preferably optionally substituted C 1-6 alkyl group, optionally substituted C 1-6 alkoxy group, oxo group, halogen an atom, or a nitro group.
• Ring F represents a 4- to 8-membered nitrogen-containing heterocyclic ring
• y represents an integer of 0 to 4
• each of y R 3g independently represents an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group
• y is an integer of 2 or more
• two adjacent R 3g may together with ring F form an optionally substituted bicyclic condensed nitrogen-containing heterocyclic ring.
• y in the formula (r17′) is 0-4.
• "y is 0" means that R3g is absent.
• the y R 3g in the formula (r17′) are each independently preferably an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom, or Two nitro groups or adjacent R 3g preferably together with ring F form an optionally substituted bicyclic fused nitrogen-containing heterocycle.
• the bicyclic condensed nitrogen-containing heterocyclic ring preferably has 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 an isoindole ring optionally substituted with, more preferably 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 It is an isoindole ring optionally substituted with 1 to 4 substituents selected from the group.
• Ring F is preferably a pyrrolidine ring or selected from the group consisting of optionally substituted alkyl groups, optionally substituted alkoxy groups and electron-withdrawing groups together with two adjacent R 3g forms an isoindole ring optionally substituted with 1 to 4 substituents.
• Ring F is more preferably a pyrrolidine ring, or together with two adjacent R 3g , optionally substituted C 1-6 alkyl group, optionally substituted C 1-6 alkoxy group, halogen It forms an isoindole ring optionally substituted with 1 to 4 substituents selected from the group consisting of atoms and nitro groups.
• Suitable compounds (Ig') include the following compounds. Each of the following compounds may be used alone or in combination of two or more.
• Ring F is a pyrrolidine ring
• y is an integer from 0 to 4
• y R 3g are each independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group
• R 2g is an optionally substituted heteroaryl group
• Ring F is a pyrrolidine ring
• y is an integer from 0 to 4
• y R 3g are each independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group
• R 2g is an optionally substituted 5 or a compound that is a 6-membered monocyclic nitrogen-containing heteroaryl group, or an optionally substituted 8- to 14-membered condensed polycyclic nitrogen-containing heteroaryl group (herein referred to as "compound (Ig-ii -a)”).
• Ring F In the formula (Ig'), Ring F, together with two adjacent R 3g , 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 forming an optionally substituted isoindole ring, and R 2g is an optionally substituted 5- or 6-membered monocyclic nitrogen-containing heteroaryl group, or an optionally substituted 8- to 14-membered is a condensed polycyclic nitrogen-containing heteroaryl group (referred to herein as “compound (Ig-ii-b)”).
• Ring F is a pyrrolidine ring
• y is an integer from 0 to 4
• y R 3g are each independently an optionally substituted alkyl group, an optionally substituted alkoxy group, or an electron-withdrawing group
• 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
• a compound that is a benzothiazolyl group referred to herein as “compound (Ig-iii-a)”).
• Ring F together with two adjacent R 3g , 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 forming an optionally substituted isoindole ring
• R 2g is (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) 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
• a compound that is a benzothiazolyl group referred to herein as “compound (Ig-iii-b)”).
• Ring F is a pyrrolidine ring
• y is an integer from 0 to 4
• R 3g are each independently an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom, or a nitro group
• R 2g is (1) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom and a nitro group
• R 2g is (1) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom and a nitro group
• Ring F is 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; forming an isoindole ring optionally substituted with 1 to 4 substituents, and R 2g is (1) substituted with 1 to 4 substituents selected from the group consisting of an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom and a nitro group; or (2) an optionally substituted C 1-6 alkyl group, an optionally substituted C 1-6 alkoxy group, a halogen atom, and a nitro group
• a compound that is a 2-benzothiazolyl group optionally substituted with 1 to 4 selected substituents referred to herein as “compound (Ig-iv-b)”.
• Preferred specific examples of the compound (Ig) include 1-(2-pyridinylthio)-2,5-pyrrolidinedione represented by the following formula (Ig-1), 1- (2-benzothiazolylthio)-2,5-pyrrolidinedione, 2-(2-pyridinylthio)-1H-isoindole-1,3(2H)-dione represented by the following formula (Ig-3), and Examples include 2-(2-benzothiazolylthio)-1H-isoindole-1,3(2H)-dione represented by the following formula (Ig-4).
• compound (I) is preferably compound (Ia) and/or compound (Ib).
• compound (Ic) is preferred.
• compound (I) is preferably compound (Ib) and/or compound (Ic), more preferably compound (Ib-i) and/or compound (Ic-i) is more preferably compound (Ib-ii) and/or compound (Ic-ii), still more preferably compound (Ib-iii) and/or compound (Ic-iii), still more preferably Compound (Ib-iv) and/or compound (Ic-iv), particularly preferably compound (Ib-v) and/or compound (Ic-iv).
• compound (I) is preferably compound (Ib-i') and/or compound (Ic-i'), more preferably compound (Ib-ii') and/or compound (Ic-ii'), more preferably compound (Ib-iii') and/or compound (Ic-iii'), still more preferably compound (Ib-iv') and/or Compound (Ic-iv').
• 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-1)), 2-(2-benzothi
• 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-1)), 2-(2-benzothi
• 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]
• 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-1)), 2-(2-benzothi
• 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-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-1)), 2-(2-benzothi
• compound (I) is preferably 2,2′-dibenzothiazolyl disulfide (compound (Ib-1)), 2-phenylisothiazolo[5,4-b ] pyridin-3(2H)-one (compound (Ic-1)), 2-isopropylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-2)), 2-phenyl isothiazolo[4,5-c]pyridin-3(2H)-one (compound (Ic-10)), 2-phenylisothiazolo[5,4-b]quinolin-3(2H)-one (compound (Ic -11)), 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-12)), 2-phenyl-2H-1,2- benzothiazin-3(4H)-one (compound (Ib-1)), 2-phenyl-2H
• compound (I) is preferably 2,2′-dibenzothiazolyl disulfide (compound (Ib-1)), 2-phenylisothiazolo[5,4-b ] Pyridin-3(2H)-one (compound (Ic-1)), 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-12 )), 2-phenyl-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]pyridin-3(2H)-one (compound (Ic-15)) , 2-(4-methoxyphenyl)iso
• 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)), most preferably 2-phenylisothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-1)) be.
• Compound (I) may be a commercially available product or may be produced by a known method.
• compound (Ib-1) ie, 2,2'-dibenzothiazolyl disulfide, MBTS
• MBTS 2,2'-dibenzothiazolyl disulfide
• Compound (Ib) can be produced, for example, by oxidation and coupling of a thiol, as shown in the formula below.
• oxidizing agents for oxidizing compound (Ib-1s) and compound (Ib-2s) include hydrogen peroxide, iodine, oxygen, N-bromosuccinimide, N-chlorosuccinimide and the like. Only one kind of oxidizing agent may be used, or two or more kinds thereof may be used in combination.
• the amount of the oxidizing agent used is preferably 1-10 mol, more preferably 1-5 mol, per 1 mol of compound (Ib-1s) and compound (Ib-2s) in total.
• Compound (Ib-2s) is preferably used in approximately the same amount as compound (Ib-1s).
• This reaction is carried out in a suitable solvent that does not interfere with the reaction.
• suitable solvent examples 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; Aromatic solvents and the like can be mentioned.
• the amount of the solvent to be used is preferably 1-20 mL, more preferably 3-10 mL, per 1 mmol of compound (Ib-1s) and compound (Ib-2s) in total.
• 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.
• compound (Ib) can be recovered by a known method (eg, extraction, etc.).
• the recovered compound (Ib) can be purified by known means (eg, recrystallization, chromatography, etc.).
• This step is a step of synthesizing the acid chloride of compound (Ic-1s). Specifically, compound (Ic-1s) and an excess amount of thionyl chloride (e.g., 10 mol per 1 mol of compound (Ic-1s)) are added to a reactor and refluxed for 1 hour to 1 night. An acid chloride of compound (Ic-1s) is obtained.
• compound (Ic-1s) and an excess amount of thionyl chloride e.g. 10 mol per 1 mol of compound (Ic-1s)
• This step is preferably performed under an inert atmosphere (for example, under an argon atmosphere).
• an inert atmosphere for example, under an argon atmosphere.
• This step is to synthesize compound (Ic-2s) by reacting an acid chloride of compound (Ic-1s) with an appropriate amine (R 4c —NH 2 ). Specifically, the solid containing the acid chloride of compound (Ic-1s) obtained in step A1, an appropriate amine (R 4c —NH 2 ) and a base are added to an appropriate solvent and reacted to obtain compound (Ic). -2s) is obtained.
• the base may be either an organic base or an inorganic base, preferably triethylamine.
• the amounts of R 4c -NH 2 and the base to be used are each preferably about 1 to 1.5 mol per 1 mol of the acid chloride of compound (Ic-1s).
• This step is performed in an appropriate solvent that does not inhibit the reaction.
• the solvent include polar ether solvents such as 1,4-dioxane and tetrahydrofuran (THF), with THF being preferred.
• THF tetrahydrofuran
• the amount of the solvent to be used is not particularly limited as long as it can dissolve the solid, but it is preferably about 5 to 10 mL per 1 mmol of the acid chloride of compound (Ic-1s).
• This step is preferably performed at a low temperature (for example, about 0°C). Completion of the reaction can be confirmed by TLC or the like. After completion of the reaction, compound (Ic-2s) can be recovered by known means (eg concentration and extraction).
• This step is a step of synthesizing compound (Ic-3s) by reacting compound (Ic-2s) with tert-butylthiol. Specifically, compound (Ic-3s) is obtained by reacting compound (Ic-2s) with tert-butylthiol in the presence of a base in an appropriate solvent.
• the amount of tert-butylthiol to be used is preferably about 1 to 1.5 mol per 1 mol of compound (Ic-2s).
• Examples of the base used in this step include alkali metal hydroxides such as sodium hydroxide and potassium hydroxide, with sodium hydroxide being preferred.
• the amount of the base to be used is preferably about 1 to 2 mol per 1 mol of compound (Ic-3s).
• This step is performed in an appropriate solvent that does not inhibit the reaction.
• the solvent include amide solvents such as dimethylformamide (DMF), dimethylacetamide, and N-methylpiperidone, with DMF being preferred.
• the amount of the solvent to be used is not particularly limited as long as it can dissolve the compound (Ic-2s), but it is preferably about 5 to 10 mL per 1 mmol of the compound (Ic-2s).
• the reaction temperature in this step is about room temperature (eg, 25°C), and the reaction time is, for example, 1 hour to 1 night, preferably about 3 to 20 hours.
• the compound (Ic-3s) can be recovered by known means (eg solid-liquid separation).
• This step is a step of synthesizing compound (Ic) in which Y 1c is carbonyl by a ring closure reaction of compound (Ic-3s). This step can be carried out by two methods of steps A4-1 and A4-2 below.
• Step A4-1 In this step, compound (Ic-3s), dimethylsulfoxide (DMSO), and chlorotrimethylsilane (TMSCl) are added to an appropriate solvent, and the resulting reaction mixture is reacted at about room temperature (eg, 25° C.). , Y 1c is a carbonyl.
• the amount of DMSO used is preferably about 1-2 mol per 1 mol of compound (Ic-3s).
• the amount of TMSCl to be used is preferably about 1 to 1.5 mol per 1 mol of compound (Ic-3s).
• This step is performed in an appropriate solvent that does not inhibit the reaction.
• the solvent include halogen-based solvents such as chloroform, dichloromethane, and 1,2-dichloroethane, with dichloromethane being preferred.
• the amount of the solvent to be used is not particularly limited as long as the compound (Ic-3s) can be dissolved therein, but it is preferably about 2 to 5 mL per 1 mmol of the compound (Ic-3s).
• the reaction time of this step is, for example, 1 hour to 1 night, preferably about 5 to 10 hours.
• the compound (Ic) wherein Y1c is carbonyl can be recovered by known means (eg solid-liquid separation).
• the recovered compound (Ic) can be purified by known means (eg, recrystallization, chromatography, etc.).
• Step A4-2 In this step, compound (Ic-3s) and meta-chloroperbenzoic acid (mCPBA) are reacted in an appropriate solvent to obtain a sulfoxide intermediate, which is then heated to give a compound in which Y 1c is carbonyl. Synthesize (Ic).
• the amount of mCPBA used is preferably about 1 to 1.5 mol per 1 mol of compound (Ic-3s).
• the reaction with compound (Ic-3s) and meta-chloroperbenzoic acid (mCPBA) is carried out in an appropriate solvent that does not inhibit the reaction.
• the solvent include halogen-based solvents such as chloroform, dichloromethane, and 1,2-dichloroethane, with dichloromethane being preferred.
• the amount of the solvent to be used is not particularly limited as long as the compound (Ic-3s) can be dissolved therein, but it is preferably about 8 to 15 mL per 1 mmol of the compound (Ic-3s).
• the reaction with compound (Ic-3s) and meta-chloroperbenzoic acid (mCPBA) is preferably carried out at a low temperature (eg, around 0°C).
• the reaction time is, for example, about 30 minutes to 1 hour.
• the sulfoxide intermediate can be recovered by known means (eg, extraction followed by concentration and recrystallization, etc.).
• Compound (Ic) wherein Y1c is carbonyl can be synthesized by heating the recovered sulfoxide intermediate in a suitable solvent that does not inhibit the cyclization reaction.
• suitable 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 (eg, recrystallization, chromatography, etc.).
• the compound (Ic-2s) (ie, an intermediate for synthesizing compound (Ic) wherein Y1c is carbonyl) can also be produced, for example, by the steps shown below.
• Step A2' This step is a step of synthesizing compound (Ic-2s) by reacting compound (Ic-1s) with a suitable amine (R 4c —NH 2 ) using a condensing agent.
• Condensing agents used in this step include, for example, dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC) and its hydrochloride (EDC ⁇ HCl), (benzotriazol-1-yloxy) tripyrrolidinophosphonium hexafluorophosphate (PyBop), O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoro Borate (TBTU), 1-[bis(dimethylamino)methylene]-5-chloro-1H-benzotriazolium-3-oxide hexafluorophosphate (HCTU), O-benzotriazole-N,N,N',N '-tetramethyluronium hexafluorophosphate (HBTU)
• R 4c —NH 2 and the condensing agent to be used are each preferably about 1 to 5 mol per 1 mol of compound (Ic-1s).
• a base may be used in this step.
• the base may be either an organic base or an inorganic base, preferably triethylamine.
• a base When a base is used, its amount is preferably about 1 to 5 mol per 1 mol of compound (Ic-1s).
• This step is performed in an appropriate solvent that does not inhibit the reaction.
• the solvent include polar ether solvents such as 1,4-dioxane and tetrahydrofuran (THF), with THF being preferred.
• the amount of the solvent to be used is not particularly limited, but is preferably about 1 to 10 mL per 1 mmol of compound (Ic-1s).
• the reaction temperature in this step is about room temperature (eg, 25°C), and the reaction time is, for example, 1 hour to 1 night, preferably about 3 to 20 hours.
• This step is preferably performed under an inert atmosphere (for example, under an argon atmosphere). Completion of the reaction can be confirmed by TLC or the like.
• compound (Ic-2s) can be recovered by known means (eg concentration and extraction).
• oligonucleotide precursor having a phosphite ester bond or a phosphonate ester bond (hereinafter referred to as "oligonucleotide (sometimes abbreviated as "precursor") is oxidized.
• the amount of compound (I) relative to the total oxidizing agent used in the production method of the present invention is preferably 80 mol% or more, more preferably 90 mol% or more.
• the oxidizing agent more preferably comprises compound (I).
• the amount of compound (I) used is preferably 1-15 mol, more preferably 1-10 mol, and still more preferably 1-5 mol, per 1 mol of the oligonucleotide precursor.
• the oligonucleotide precursor is preferably (1) an oligonucleotide precursor having a phosphite ester bond obtained by condensation of a nucleoside, nucleotide or oligonucleotide with a phosphoramidized nucleoside, nucleotide or oligonucleotide, or (2) an oligonucleotide and an oligonucleotide precursor having a phosphite ester bond obtained by condensation with a phosphoramidite.
• the oligonucleotide precursor of (1) for example, the compound (cI) (that is, obtained by condensation of the compound (aI) and the compound (bI), phosphorous acid an oligonucleotide precursor having an ester bond), and the compound (cI') (that is, phosphorous acid obtained by condensation of the compound (aI') and the compound (bI') oligonucleotide precursors having an ester bond).
• the oligonucleotide precursor of (2) include the compound ( ⁇ -I) and the compound ( ⁇ -I').
• Oligonucleotide precursors with phosphite ester linkages are more preferably obtained by condensation of nucleosides, nucleotides or oligonucleotides with phosphoramidated nucleosides, nucleotides or oligonucleotides, phosphite ester linkages is an oligonucleotide precursor having
• the oligonucleotide precursor is more preferably a nucleoside, nucleotide or oligonucleotide (a) having a hydrophobic protecting group, a hydroxyl group protected with a temporary protecting group removable under acidic conditions, and a phosphoramidite nucleosides, nucleotides or oligonucleotide precursors with phosphite linkages obtained by condensation with oligonucleotides (b).
• the oligonucleotide precursor is particularly preferably the compound (cI) in which Pg is **LYZ (that is, the compound (aI) in which Pg is **LYZ) and Oligonucleotide precursor having a phosphite ester bond obtained by condensation with the compound (bI)), or the compound (cI') wherein Pg is **LYZ (i.e. , an oligonucleotide precursor having a phosphite ester bond, obtained by condensation of the compound (aI′) and the compound (bI′), wherein Pg is **LYZ. be.
• the oligonucleotide precursor is preferably an oligonucleotide precursor having a thiophosphate ester bond in addition to a phosphite ester bond or a phosphonate ester bond.
• an oligonucleotide precursor having a thiophosphate ester bond is oxidized with iodine, a by-product (desulfurized product) obtained by converting the thiophosphate ester bond to a phosphate ester bond is produced. According to the present invention using , the generation of this desulfurized product can be suppressed.
• Water present in the system acts as a source of oxygen atoms and thus can promote oxidation of the oligonucleotide precursor.
• the amount of water is preferably 0.5 to 200 mol, more preferably 1 to 100 mol, still more preferably 1 to 50 mol, per 1 mol of the oligonucleotide precursor.
• the reaction temperature for 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. be.
• the oxidation of the oligonucleotide precursor is preferably carried out in a solution containing a non-polar solvent.
• 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.
• nonpolar solvents examples include halogen 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 pentane, heptane, octane, nonane and cyclohexane; and non-polar ether solvents such as diethyl ether, cyclopentyl methyl ether and tert-butyl methyl ether. Only one type of nonpolar solvent may be used, or two or more types may be used in combination.
• halogen solvents such as chloroform, dichloromethane, and 1,2-dichloroethane
• aromatic solvents such as benzene, toluene, xylene, and mesitylene
• ester solvents such as ethyl
• the non-polar solvent is preferably at least one selected from the group consisting of halogen solvents, aromatic solvents, ester solvents and aliphatic solvents, more preferably halogen solvents and toluene, still more preferably It is at least one selected from the group consisting of chloroform, dichloromethane and toluene, and particularly preferably dichloromethane.
• a solution containing a non-polar solvent may further contain an aprotic polar solvent.
• the aprotic polar solvent include nitrile solvents such as acetonitrile and propionitrile; polar ether solvents such as tetrahydrofuran; and pyridine. Among these, nitrile solvents are preferred, and acetonitrile and pyridine are more preferred.
• the amount of the aprotic polar solvent used is preferably 10-100 mL, more preferably 10-50 mL, per 100 mL of the non-polar solvent.
• the production method of the present invention can also be carried out by one-pot synthesis.
• the one-pot synthesis is In a solution containing a non-polar solvent, a nucleoside, nucleotide or oligonucleotide (a) having a hydrophobic protecting group, and a hydroxyl group protected with a temporary protecting group removable under acidic conditions, and phosphoramidated condensed with a nucleoside, nucleotide or oligonucleotide (b), having a phosphite ester bond and a hydrophobic protecting group, and a hydroxyl group protected with a temporary protecting group removable under acidic conditions step (1) of forming an oligonucleotide precursor (c);
• a quenching agent (i) for the phosphoramidated nucleoside, nucleotide or oligonucleotide (b) is added to the solution after step (1) to quench the phosphoramidated nucleoside, nucleotide
• step (3) of forming an oligonucleotide (d) protected with a temporary protecting group step (4) of adding a quenching agent (ii) for the oxidizing agent to the solution after step (3) to quench the oxidizing agent; adding an acid to the solution after step (4) to remove the temporary protecting group removable under acidic conditions of the oligonucleotide (d), resulting in an oligonucleotide having an unprotected hydroxyl group and a hydrophobic protecting group; step (5) of forming (e); If necessary, a step (6) of adding a base to the solution after step (5), and a step of adding a polar solvent to the solution containing the oligonucleotide (e) to precipitate the oligonucleotide (e).
• the oxidizing agent is compound (I) (excluding 2,2-dipyridyl disulfide).
• intermediates that is, the oligonucleotide precursor (c) obtained in step (1) and the oligonucleotide (d) obtained in step (3)
• step (1) the oligonucleotide precursor (c) obtained in step (1)
• step (3) the oligonucleotide (d) obtained in step (3)
• Step (1) (condensation)
• a nucleoside, nucleotide or oligonucleotide (a) having a hydrophobic protecting group and a hydroxyl group are protected with a temporary protecting group removable under acidic conditions.
• a group-protected oligonucleotide precursor (c) is formed.
• 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.
• the temporary protective group for the hydroxyl group of the phosphoramidized nucleoside, nucleotide or oligonucleotide (b) is preferably a 4,4'-dimethoxytrityl group or a 4-mono It is a methoxytrityl group, more preferably a 4,4'-dimethoxytrityl group.
• the combination of said nucleoside, nucleotide or oligonucleotide (a) used in step (1) and said phosphoroamidated nucleoside, nucleotide or oligonucleotide (b) is preferably combined with said compound (aI) A combination with the compound (bI), or a combination of the compound (aI') and the compound (bI'), more preferably Pg is **LYZ a combination of compound (aI) and said compound (bI), or said compound (aI') wherein Pg is **L-Y-Z and said compound (b-I') is a combination of
• Step (1) is performed in a solution containing a non-polar solvent.
• nonpolar solvents include halogen 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 pentane, heptane, octane, nonane and cyclohexane; and non-polar ether solvents such as diethyl ether, cyclopentyl methyl ether and tert-butyl methyl ether.
• nonpolar solvent is preferably at least one selected from the group consisting of halogen solvents, aromatic solvents, ester solvents and aliphatic solvents, more preferably selected from the group consisting of halogen solvents and toluene. At least one, more preferably at least one selected from the group consisting of chloroform, dichloromethane and toluene, particularly preferably dichloromethane and/or toluene.
• the steps after step (1) are similarly carried out in a solution containing a non-polar solvent.
• a solution containing a non-polar solvent may further contain an aprotic polar solvent.
• the aprotic polar solvent 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 the aprotic polar solvent used is preferably 10-100 mL, more preferably 10-50 mL, per 100 mL of the non-polar solvent.
• the amount of the phosphoramidized nucleoside, nucleotide or oligonucleotide (b) used is, for example, 1 to 10 mol, preferably 1 to 5 mol, per 1 mol of the nucleoside, nucleotide or oligonucleotide (a) used. is.
• the concentration of said nucleosides, nucleotides or oligonucleotides (a) in solution is preferably 1-30% by weight.
• the reaction temperature in step (1) is not particularly limited as long as the 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., and is, for example, 5 minutes to 24 hours.
• a known activator may be used to promote condensation.
• activating agents include pyridine trifluoroacetate, tetrazole, 5-ethylthio-1H-tetrazole, 5-benzylthio-1H-tetrazole, 4,5-dicyanoimidazole and the like. 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). be.
• Step (2) Quenching phosphoroamidated nucleoside, nucleotide or oligonucleotide (b))
• a quenching agent (i) of the phosphoramidated nucleoside, nucleotide or oligonucleotide (b) is added to the solution after step (1) to remove the phosphoramidated , nucleosides, nucleotides or oligonucleotides (b).
• the quenching agent (i) may be used alone or in combination of two or more. Quenching agents (i) include, for example, water, alcohols, phenols and amines.
• Alcohols that can be used as quenching agent (i) include, for example, methanol, 2-propanol, t-butanol, 2,2,2,-trifluoroethanol, tetrahydrofurfuryl alcohol, furfuryl alcohol, 2,3- optionally halogenated monohydric alcohols such as O-isopropylidene-D-ribofuranose and 3′-O-triisopropylsilyl-thymidine; and optionally halogenated polyhydric alcohols such as ethylene glycol and diethylene glycol. mentioned.
• Phenols that can be used as the quenching agent (i) include, for example, 4-nitrophenol and pentafluorophenol.
• Amines that can be used as quenching agents (i) include, for example, morpholine.
• the amount of quenching agent (i) used is preferably 1 to 20 mol, more preferably 1 ⁇ 10 mol, more preferably 1-5 mol.
• the quenching agent (i) is preferably water.
• the subsequent oxidation in step (3) can be facilitated by using water as quenching agent (i).
• the amount used is preferably 1 to 1 mol of the phosphoramidized nucleoside, nucleotide or oligonucleotide (b) used in step (1). 20 mol, more preferably 2 to 15 mol, still more preferably 2 to 10 mol.
• the temperature of the solution after addition of the quenching agent (i) is not particularly limited as long as the phosphoramidized nucleoside, nucleotide or oligonucleotide (b) can be quenched, preferably 5 to 40° C., more preferably. is 15-30°C.
• the stirring time of the solution after addition of quenching agent (i) varies depending on the type of quenching agent (i) used, temperature, etc., but is, for example, 10 minutes to 3 hours.
• step (3) (oxidation)
• compound (I) excluding 2,2-dipyridyl disulfide
• step (3) compound (I) (excluding 2,2-dipyridyl disulfide) is used as an oxidizing agent to oxidize the phosphite ester bond of the oligonucleotide precursor (c) to form phosphoric acid. It is characterized by conversion to an ester bond.
• step (3) an oxidizing agent (i.e., compound (I)) is added to the solution after step (2) to oxidize the oligonucleotide precursor (c), resulting in a phosphate ester bond and An oligonucleotide (d) is formed which has a hydrophobic protecting group and whose hydroxyl group is protected with a temporary protecting group that can be removed under acidic conditions. Descriptions of oxidizing agents and oxidation are provided above.
• the oligonucleotide precursor (c) is preferably an oligonucleotide precursor having a thiophosphate ester bond in addition to a phosphite ester bond.
• Aromatic amine is preferably added to the solution after step (3).
• Aromatic amines may be used alone or in combination of two or more. Addition of an aromatic amine can further suppress the formation of defect bodies.
• the addition of the aromatic amine may be 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). Alternatively, quenching agent (ii) and aromatic amine may be added sequentially in step (4).
• aromatic amines examples include aniline, 2-chloroaniline, 3-chloroaniline, 2,4-dichloroaniline, 2-fluoroaniline, 4-methoxyaniline, 4-nitroaniline, 2,6-dichloroaniline, 2 , 6-xylidine and the like.
• Aromatic amines are preferably aniline, 2-chloroaniline, 3-chloroaniline, 2,4-dichloroaniline, 2-fluoroaniline, 4-methoxyaniline, 4-nitroaniline, 2,6-dichloroaniline, 2, At least one selected from the group consisting of 6-xylidine, more preferably 2-chloroaniline and/or 2,6-xylidine, still more preferably 2,6-xylidine (that is, 2,6-dimethylaniline ).
• the amount of aromatic amine used is preferably 1 to 20 mol, more preferably 1 to 10 mol, and still more preferably 1 to 5 mol, per 1 mol of the oxidizing agent used in step (3). be.
• Step (4) quenching of oxidizing agent
• the oxidizing agent quenching agent (ii) is added to the solution after step (3) to quench the oxidizing agent.
• the quenching agent (ii) may be used alone or in combination of two or more.
• the quenching agent (ii) is preferably an organophosphorus compound. Only one type of organic phosphorus compound may be used, or two or more types may be used in combination.
• the organic phosphorus compound is preferably at least one selected from the group consisting of phosphines, phosphite triesters, phosphite esters, phosphonite diesters and phosphinate esters, and more preferably phosphines.
• phosphines phosphite triesters, phosphite esters, phosphonite diesters and phosphinate esters, and more preferably phosphines.
• phosphines and the like may be used alone or in combination of two or more.
• Phosphines are compounds represented by the formula: P(R) 3 (wherein the three R's each independently represent a hydrogen atom, an alkyl group or an aryl group). Phosphines include, for example, triphenylphosphine and methyldiphenylphosphine.
• a phosphite triester is a compound represented by the formula: P(OR') 3 (wherein the three R's each independently represent an alkyl group or an aryl group).
• Phosphite triesters include, for example, triethyl phosphite.
• Phosphite esters have the formula: P(OR')(R) 2 (wherein the two R's each independently represent a hydrogen atom, an alkyl group or an aryl group, and R' is an alkyl group or an aryl It is a compound represented by ).
• Phosphite esters include, for example, ethoxydiphenylphosphine and the like.
• Phosphonite diesters have the formula: P(R)(OR') 2, where R represents a hydrogen atom, an alkyl group or an aryl group, and two R' are each independently an alkyl group or (representing an aryl group).
• Examples of phosphonous diesters include diethoxyphenylphosphine and the like.
• Phosphinate esters include, for example, 9,10-dihydro-9-oxa-10-phosphaphenanthrene 10-oxide represented by the following formula.
• the quenching agent (ii) is preferably triphenylphosphine, methyldiphenylphosphine, triethyl phosphite, ethoxydiphenylphosphine, diethoxyphenylphosphine, and 9,10-dihydro-9- It is at least one selected from the group consisting of oxa-10-phosphaphenanthrene 10-oxide, more preferably triphenylphosphine and/or methyldiphenylphosphine, still more preferably triphenylphosphine.
• the amount of the quenching agent (ii) to be used is sufficient as long as it can quench the surplus of the oxidizing agent in the step (3). It is preferably 1 to 10 mol, more preferably 1 to 5 mol, per 1 mol of phosphoramidized nucleoside, nucleotide or oligonucleotide (b) used in step (1)).
• the temperature of the solution after addition of the quenching agent (ii) is not particularly limited as long as the oxidizing agent can be quenched, but is preferably 0°C to 50°C, more preferably 10°C to 40°C.
• the stirring time of the solution after addition of 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.
• Step (5) (removal of temporary protecting group)
• an acid is added to the solution after step (4) to remove temporary protecting groups removable under acidic conditions of said oligonucleotide (d), resulting in unprotected hydroxyl groups and hydrophobic
• An oligonucleotide (e) with protecting groups is formed. Only one type of acid may be used, or two or more types may be used in combination.
• the acid is not particularly limited as long as it can remove the temporary protecting group well, and examples thereof include trifluoroacetic acid, dichloroacetic acid, trifluoromethanesulfonic acid, trichloroacetic acid, methanesulfonic acid, hydrochloric acid, acetic acid, p-toluenesulfonic acid and the like. .
• trifluoroacetic acid, dichloroacetic acid, trifluoromethanesulfonic acid and trichloroacetic acid are more preferred, trifluoroacetic acid, dichloroacetic acid and trifluoromethanesulfonic acid are more preferred, trifluoroacetic acid and trifluoromethane Sulfonic acids are even more preferred, and trifluoroacetic acid is particularly preferred.
• the amount of acid used is, for example, 1 to 100 mol, preferably 1 to 40 mol, per 1 mol of oligonucleotide (d).
• 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, the type of nonpolar solvent, the reaction temperature, etc., and is, for example, 5 minutes to 5 hours.
• a cation scavenger is preferably added to the solution before, during or after 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 removing the temporary protecting group. Only one kind of cation scavenger may be used, or two or more kinds thereof may be used in combination.
• the cation scavenger is not particularly limited as long as the removed temporary protecting group does not reprotect or undergo a side reaction to the deprotected functional group, but pyrrole, 2-methylpyrrole, 3-methylpyrrole, 2 ,3-dimethylpyrrole, 2,4-dimethylpyrrole and other pyrrole derivatives; , 6,7-dimethylindole, 5-methoxyindole, and other indole derivatives; 2-methylfuran, 2,3-dimethylfuran, 2-methyl-3-(methylthio)furan, menthofuran, and other furan derivatives. can be done.
• the amount of the cation scavenger to be used is preferably 1-50 mol, more preferably 5-20 mol, per 1 mol of the oligonucleotide (d).
• Step (6) neutralization
• the one-pot synthesis includes step (6) of adding a base to the solution after step (5) after step (5) and before step (7). ) may be further included.
• step (7) solid-liquid separation
• step (6) neutralization
• 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, 5-nitrobenzimidazole and the like.
• pyridine, 2,4,6-trimethylpyridine, benzimidazole, 1,2,4-triazole, N-phenylimidazole, N-methylimidazole, 2-amino-4,6-dimethylpyrimidine, 1, 10-phenanthroline is preferred
• pyridine, 2,4,6-trimethylpyridine, benzimidazole, 1,2,4-triazole, N-phenylimidazole are more preferred
• pyridine, 2,4,6-trimethylpyridine, benzimidazole, 1,2,4-triazole is more preferred
• pyridine, 2,4,6-trimethylpyridine and benzimidazole are particularly preferred.
• the amount of base used in step (6) is preferably 1 to 10 mol, more preferably 1 to 3 mol, per 1 mol of acid used in step (5).
• Step (7) solid-liquid separation
• a polar solvent is added to the solution containing the oligonucleotide (e) (i.e., the solution after step (5) or, if necessary, the solution after step (6)) to obtain the oligonucleotide (e) is deposited.
• Polar solvents used in step (7) include, for example, alcohol solvents such as methanol, ethanol and isopropanol; nitrile solvents such as acetonitrile and propionitrile; ketone solvents such as acetone and 2-butanone; polar ether solvents such as dioxane and tetrahydrofuran; amide solvents such as dimethylformamide, dimethylacetamide and N-methylpiperidone; sulfoxide solvents such as dimethylsulfoxide; A polar solvent may use only 1 type and may use 2 or more types together. Among these, nitrile solvents are preferable, and acetonitrile is more preferable.
• the amount of the polar solvent added in the solid-liquid separation is preferably 1 to 20 mL, more preferably 5 to 20 mL, per 1 mL of the nonpolar solvent contained in the solution. , more preferably 5-10 mL.
• a precipitation accelerator for example, 3,4,5-tris(octadecyloxy)benzylpivalate
• WO 2016/117663 a precipitation accelerator
• the precipitated oligonucleotide (e) can be recovered by known means such as filtration.
• the oligonucleotide chain can be extended by repeating the one-pot synthesis including steps (1) to (5) and (7) (preferably steps (1) to (7)).
• a method for producing an oligonucleotide by repeating such one-pot synthesis is also included in the production method of the present invention.
• the protecting group eg, phosphate group protecting group, hydrophobic protecting group
• the resulting oligonucleotide (e) is removed by a known method.
• the present invention provides novel compounds of the formula (In):
• R 1n represents an optionally substituted phenyl group
• p′ represents 0, ring C′′ represents a nitrogen atom-containing 10-membered bicyclic condensed aromatic heterocycle
• q ' indicates 0,
• R 1n represents an optionally substituted phenyl group
• p′ represents 0, ring C′′ represents a 6-membered nitrogen-containing aromatic heterocycle
• q′ represents 1
• R 3n represents a C 1-6 perfluoroalkyl group
• R 1n represents an optionally substituted phenyl group
• p′ represents 1, ring C′′ represents a 6-membered nitrogen-containing aromatic heterocycle
• q′ represents 0,
• R 1n represents a pyridyl group substituted with one C 1-6 alkyl group
• p′ represents 0, ring C′′ represents a 6-membered nitrogen-containing aromatic heterocyclic ring
• q ' indicates 0.
• a compound represented by is also provided.
• the compound (In) is oxidized to oxidize an oligonucleotide precursor having a phosphite ester bond or a phosphonate ester bond (particularly, an oligonucleotide precursor having a phosphite ester bond). useful as an agent. Only one compound (In) may be used, or two or more thereof may be used in combination.
• the "optionally substituted phenyl group” of R 1n is preferably a phenyl group.
• the “pyridyl group substituted with 1 C 1-6 alkyl group” of R 1n is preferably a pyridyl group substituted with 1 methyl group, more preferably 4-methyl-2-pyridyl is the base.
• the "nitrogen atom-containing 10-membered bicyclic fused aromatic heterocycle" of ring C" is preferably a quinoline ring.
• the "6-membered nitrogen-containing aromatic heterocycle” of Ring C" is preferably a pyridine ring.
• the “C 1-6 perfluoroalkyl group” of R 3n is preferably a trifluoromethyl group.
• Compound (In) is preferably 2-phenylisothiazolo[5,4-b]quinolin-3(2H)-one (compound (Ic-11)), 2-phenyl-5-trifluoromethylisothiazolo[5 ,4-b]pyridin-3(2H)-one (compound (Ic-12)), 2-phenyl-2H-1,2-benzothiazin-3(4H)-one (compound (Ic-13)), or 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridin-3(2H)-one (compound (Ic-15)).
• Compound (In) can be prepared by the above-mentioned "method for producing compound (Ic) wherein Y 1c is carbonyl", synthesis examples described later, known methods (e.g., the method described in Supporting Information of Non-Patent Document 2), or It can be manufactured by the method according to.
• oligonucleotide HO-T-SUC-TOB: deoxythymidin-3′-yl 3,4,5-tris(octadecyloxy)benzylsuccinate
• HO-T(S)T-SUC-TOB deoxythymidine-3′-[O -(2-cyanoethyl)]phosphorothionyl deoxythymidin-3′-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate
• oligonucleotide 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)]phosphityl 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 deoxythymidin-3'-yl-[3,4,5-tris(octadecyloxy)benzyl]succinate DMT
• Synthesis Example 1 Synthesis of oligonucleotide precursor having a phosphite ester bond . 5.0 mmol), dehydrated dichloromethane (100 mL) and dehydrated acetonitrile (30 mL) were added to prepare a solution. dT-CE phosphoramidite (3.0 g, 4.0 mmol) and triphenylphosphine (270 mg, 1.0 mmol) were then 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.
• 2,2,2-Trifluoroethanol (1.5 mL, 20 mmol) was then added to the solution and stirred at room temperature for 30 minutes, followed by 2,6-dimethylaniline (1.8 mL, 15 mmol) and POS (1.0 g). , 5.1 mmol) was added and the solution was stirred at room temperature for 1.0 hour. After that, 2,3-dimethylfuran (2.1 mL, 20 mmol), trifluoroacetic acid (5.2 mL, 68 mmol), and 2,6-dimethylaniline (83.5 ⁇ L, 0.7 ⁇ mol) were sequentially added, and the solution was brought to room temperature. and stirred for 55 minutes.
• Example 1 Compound (Ib-1) was added to a 20 mL two-necked flask under an argon oxide atmosphere, and the solid obtained in Synthesis Example 1 (100.0 mg: DMTrO-G (III) A (S) C ( S)T(S)T-SUC-TOB (56.9 mg, 17.4 ⁇ mol), dehydrated dichloromethane (0.9 mL) and dehydrated acetonitrile (0.3 mL) were added to prepare a solution. Water (9 ⁇ L, 0.5 mmol) and compound (Ib-1) (12.1 mg, 36.4 ⁇ mol) were then sequentially added, and the solution was stirred at room temperature for 60 minutes.
• Triphenylphosphine (4.6 mg, 18 ⁇ mol) was then added and the solution was stirred for 30 minutes. Subsequently, acetonitrile (10 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and Piv-TOB and the pentamer oligonucleotide DMTrO-G(O)A. A mixture of (S)C(S)T(S)T-SUC-TOB was obtained (82 mg, 82% yield). In addition, the compound (Ib-1) used the commercial item. m/z: Calcd. 3277.48, Found 1639.75 [M+2H] 2+
• Example 2 Compound (Ic-1) was added to a 20 mL two-necked flask under an argon oxide atmosphere, and the solid obtained in Synthesis Example 1 (100 mg: DMTrO-G(III)A(S)C(S)T (57 mg as (S)T-SUC-TOB, 17 ⁇ mol), dehydrated dichloromethane (0.9 mL), and dehydrated acetonitrile (0.3 mL) were added to prepare a solution. Water (9 ⁇ L, 0.5 mmol) and compound (Ic-1) (8.0 mg, 35 ⁇ mol) were then sequentially added and the solution was stirred at room temperature for 60 minutes.
• Triphenylphosphine (4.7 mg, 18 ⁇ mol) was then added and the solution was stirred for 30 minutes. Subsequently, acetonitrile (10 mL) was added, the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and Piv-TOB and the pentameric oligonucleotide DMTrO-G(O)A(S ) to give a mixture of C(S)T(S)T-SUC-TOB (83 mg, 83% yield).
• the compound (Ic-1) used was synthesized by the method described in Supporting Information of Non-Patent Document 2. m/z: Calcd. 3277.48, Found 1639.75 [M+2H] 2+
• Triphenylphosphine (4.8 mg, 18 ⁇ mol) was then added and the solution was stirred for 30 minutes. Subsequently, acetonitrile (10 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel, dried, and Piv-TOB and the pentameric oligonucleotide DMTrO-G(O)A ( A mixture of S)C(S)T(S)T-SUC-TOB was obtained (83 mg, 83% yield). m/z: Calcd. 3277.48, Found 1639.75 [M+2H] 2+
• Example 1 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, heated at 65° C. for 4 hours, and then cooled to room temperature. After removing insoluble matter in the reaction solution with a syringe filter, it was concentrated under reduced pressure with a centrifugal evaporator, and the oligonucleotide DMTrO-G(O)A(S)C(S)T(S)T-OH having a thiophosphate ester bond was obtained. (hereinafter referred to as "the oligonucleotide of interest”) was obtained. m/z: Calcd. 1827.36, Found 1826.34 [MH] -
• the resulting mixture containing the oligonucleotide of interest and the desulfurized product was analyzed by liquid chromatography-mass spectrometry (LC-MS).
• LC-MS liquid chromatography-mass spectrometry
• EIC extracted ion chromatogram
• Percentage of desulfurized product (%) (peak area of desulfurized product/peak area of target oligonucleotide) x 100
• the ratio (%) of the desulfurized product was calculated. Table 1 shows the results.
• Synthesis Example 2 Synthesis of Oligonucleotide Precursor Having Phosphonate Ester Bond HO-T-SUC-TOB (500 mg, 400 ⁇ mol), dT-H-phosphonate TEA salt (568 mg, 800 ⁇ mol) in a 200 mL three-necked flask under an argon atmosphere. , anhydrous dichloromethane (10 mL), and anhydrous pyridine (10 mL) were added to form a solution. Subsequently pivaloyl chloride (0.2 mL, 1.6 mmol) was added and the solution was stirred for 1 hour.
• Example 3 Using compound (Ic-1), the oligonucleotide precursor (171 mg, 93 ⁇ mol) obtained in Synthesis Example 2, dichloromethane (4.7 mL), and acetonitrile were placed in a 50 mL two-necked flask under an oxidized argon atmosphere. (1.4 mL) was added to prepare a solution. Subsequently, water (60 ⁇ L), compound (Ic-1) (43 mg, 189 ⁇ mol) and diazabicycloundecene (DBU) (55 ⁇ L, 372 ⁇ mol) were added and the solution was stirred at room temperature for 2 hours.
• DBU diazabicycloundecene
• Example 4 Using compound (Ib-1), the oligonucleotide precursor (170 mg, 93 ⁇ mol) obtained in Synthesis Example 2, dichloromethane (4.7 mL), and acetonitrile were placed in a 50 mL two-necked flask under an oxidized argon atmosphere. (1.4 mL) was added to prepare a solution. Subsequently, compound (Ib-1) (62 mg, 186 ⁇ mol) and DBU (55 ⁇ L, 372 ⁇ mol) were added and the solution was stirred at room temperature for 2 hours.
• Non-Patent Document 1 ARKIVOC 2009 (iii) 264-273
• H-phosphonate compound 1
• a nucleoside compound 2
• compound 4 a dinucleotide
• the obtained solid (10 mg) and 28% by weight aqueous ammonia (5 mL) were placed in an autoclave, heated at 65°C for 4 hours, and then cooled to room temperature. After removing insoluble matter in the reaction solution with a syringe filter, the solution was concentrated under reduced pressure with a centrifugal evaporator, and the obtained concentrate was subjected to LC-MS analysis.
• the obtained solid (10 mg) and 28% by weight aqueous ammonia (5 mL) were placed in an autoclave, heated at 65°C for 4 hours, and then cooled to room temperature. After removing insoluble matter in the reaction solution with a syringe filter, the solution was concentrated under reduced pressure with a centrifugal evaporator, and the obtained concentrate was subjected to LC-MS analysis.
• oligonucleotide of interest The amount of the oligonucleotide HO-T(S)T(S)T-OH having an evaluation thiophosphate ester bond (hereinafter referred to as the "oligonucleotide of interest"), and the oligonucleotide of interest and the oxidizing agent (2 ,2′-dipyridyl disulfide or compound (Ib-1)) formed by reaction (hereinafter referred to as “by-product”) was measured as follows, and the amount of by-product was calculated.
• Synthesis Example 3 Synthesis of 2-(2-benzothiazolyldithio)propanoic acid (compound (Ib-2)) In a 200 mL eggplant flask, tetrahydrofuran (100 mL), 2-mercaptopropionic acid (10 mmol), and compound ( Ib-1) (20 mmol) was added and the solution was stirred for 24 hours. After that, the solution was filtered, the solvent of the filtrate was distilled off under reduced pressure, and the resulting concentrate was purified by silica gel column chromatography to obtain the desired product (1.6 g).
• Synthesis Example 4 Synthesis of 3-(2-benzothiazolyldithio)propanoic acid (compound (Ib-3)) Tetrahydrofuran (100 mL), 3-mercaptopropionic acid (10 mmol), and compound ( Ib-1) (20 mmol) was added to prepare a solution and the solution was stirred for 24 hours. After that, the solution was filtered, the solvent of the filtrate was distilled off under reduced pressure, and the resulting concentrate was purified by silica gel column chromatography to obtain the desired product (1.3 g).
• Synthesis Example 5 Synthesis of 2,2'-dibenzimidazolyl disulfide (compound (Ib-4)) Dichloromethane (150 mL), 2-mercaptobenzimidazole (20 mmol), and triethylamine (22 mmol) were added to a 300 mL round-bottom flask, and then Iodine (11 mmol) was added and the solution was stirred for 3 hours. Thereafter, the solution was filtered, the solvent of the filtrate was distilled off under reduced pressure, and the resulting concentrate was recrystallized using diethyl ether and ethanol to obtain the desired product (2.5 g).
• Synthesis Example 6 Synthesis of 2,2'-dibenzoxazolyl disulfide (compound (Ib-5)) 2-mercaptobenzoxazole (1.5 g, 10 mmol) and ethyl acetate (18 mL) were added to a 30 mL eggplant flask. , a solution was prepared. Hydrogen peroxide (33% by weight, 489 ⁇ L) was added dropwise to the solution at 0° C. over 60 minutes, then the solution was warmed 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, then the organic layer was separated and the aqueous layer was extracted twice with ethyl acetate (5 mL).
• Synthesis Example 7 Synthesis of 2,2′-dithiobis(pyridine-N-oxide) (compound (Ib-8)) Into a 100 mL round-bottomed flask, 2-mercaptopyridine-N-oxide (3.8 g, 30 mmol) and water ( 27 mL) was added to prepare a solution. Hydrogen peroxide (33% by weight, 3.2 mL) was added dropwise to the solution at 0° C. over 60 minutes, then the solution was heated to 40° C. and stirred for 1 hour. The solution was filtered to collect a solid, which was washed with methanol and dried under reduced pressure to obtain the target product (1.2 g).
• Synthesis Example 8 Synthesis of oligonucleotide precursor having a phosphite ester bond. Anhydrous acetonitrile (26 mL) was added to form a solution. Then molecular sieves 3A (1.7 g), triphenylphosphine (227 mg, 867 ⁇ mol), dT-CE phosphoramidite (2.6 g, 3.5 mmol) and 5-ethylthio-1H-tetrazole (452 mg, 3.47 mmol). were sequentially added, 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.
• Examples 5-15 and Comparative Example 2 DMTrO-C(III)T(S)T-SUC-TOB (60 mg, 25 ⁇ mol), dehydrated dichloromethane (1.3 mL) and dehydrated acetonitrile in oxidation flasks using various oxidizing agents. (0.4 mL) was added to prepare a solution. After water (17 ⁇ L) was added to the solution, various oxidizing agents (38 ⁇ mol) shown in Table 3 below were added, and the mixture was stirred at room temperature for 3 hours to oxidize the trimer oligonucleotide DMTrO—C(O). T(S)T-SUC-TOB (hereinafter referred to as "oxidant”) was synthesized.
• oxidant T(S)T-SUC-TOB
• the oligonucleotide precursor can be oxidized more efficiently than the 2,2'-dipyridyl disulfide used in .
• 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 was placed in a 300 mL two-necked flask. -TOB (351 mg, 284 ⁇ mol), Piv-TOB (350 mg, 350 ⁇ mol), dehydrated dichloromethane (14 mL) and dehydrated acetonitrile (4 mL) were added to prepare a solution.
• Triphenylphosphine (74.2 mg, 283 ⁇ mol) was then added and the solution was stirred for 30 minutes before the sequential addition of 2,3-dimethylfuran (298 ⁇ L, 2.83 mmol) and trifluoroacetic acid (563 ⁇ L, 7.35 mmol). , the solution was stirred at room temperature for 30 minutes. Further trifluoroacetic acid (86.6 ⁇ L, 1.13 mmol) was added and the solution was stirred at room temperature for 30 minutes. Pyridine (2.0 mL, 25 mmol) and water (120 ⁇ L) were then sequentially added to the solution, and the solution was stirred at room temperature for 60 minutes.
• Piv-TOB and trimer were produced in the same manner as above, except that the solid obtained above (766 mg) and 2'-OMe-C-CE phosphoramidite were used instead of dT-CE phosphoramidite.
• a mixture of oligonucleotides HO-Cm(O)T(O)T-SUC-TOB was obtained (875 mg, 98% yield). m/z: Calcd. 2070.17, Found 2071.19 [M+H] +
• Example 17 Synthesis of DMTrO-Af(O)Cm(O)T(O)T-OH using compound (Ic-1) as an oxidizing agent Compound (Ib-1) was changed to compound (Ic-1) An aqueous solution of DMTrO--Af(O)Cm(O)T(O)T--OH was obtained in the same manner as in Example 16 except for the above. m/z: Calcd. 1498.39, Found 1497.37 [MH] -
• 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 ⁇ mol) and Piv-TOB (351 mg, 352 ⁇ mol) were added, followed by dry dichloromethane (14 mL) and dry acetonitrile (4.2 mL) to prepare a solution. Then dT-CE phosphoramidite (406 mg, 545 ⁇ mol) and 5-ethylthio-1H-tetrazole (70.1 mg, 539 ⁇ mol) were added sequentially and the solution was stirred at room temperature for 45 minutes.
• Piv-TOB and trimer were produced in the same manner as above, except that the solid obtained above (812 mg) and 2'-OMe-C-CE phosphoramidite were used instead of dT-CE phosphoramidite.
• a mixture of oligonucleotides HO-Cm(O)T(O)T-SUC-TOB was obtained (893 mg, 94% yield). m/z: Calcd. 2070.17, Found 2071.19 [M+H] +
• DMTrO-Af(O)Cm(O)T(O)T-OH (hereinafter "Oligonucleotide of interest")
• DMTrO-Af(O)Cm(O)T-OH or DMTrO which is a trimer lacking the second or third residue from the oligonucleotide of interest, respectively -Af(O)T(O)T-OH (hereinafter referred to as "deficiency”) was measured by MS analysis.
• Example 18 Synthesis of HO-Af(O)Cm(O)T(O)T-SUC-TOB using compound (Ic-1) as oxidant (with addition of aromatic amine) HO-T-SUC-TOB (6.4 g, 5.2 mmol), Piv-TOB (13 g, 13 mmol), dehydrated dichloromethane (349 mL) and dehydrated acetonitrile (98 mL) were added to a 3 L three-necked flask under an argon atmosphere. , a solution was prepared.
• Triphenylphosphine 1. g, 5.2 mmol
• 2,6-xylidine 5.5 mL, 45 mmol
• 2,3-dimethylfuran 5.4 mL, 52 mmol
• trifluoroacetic acid 13.8 mL, 180 mmol
• Pyridine 43.7 mL, 541 mmol
• water 2.5 mL
• DMTrO-Af(O)Cm(O)T(O)T-SUC-TOB (3.4 mg) obtained above and 28% by weight aqueous ammonia (5 mL) were placed in an autoclave and heated at 65° C. for 4 hours. After that, it was cooled to room temperature. After removing insoluble matter in the reaction solution with a syringe filter, the solution was concentrated under reduced pressure with 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] -
• Example 17 and Example 18 DMTrO-Af(O)Cm(O)T(O)T-OH in the aqueous solution obtained in Example 17 and Example 18 (hereinafter referred to as "oligonucleotide of interest” ), and DMTrO-Af(O)Cm(O)T-OH or DMTrO-Af(O)T(O), which are trimers lacking the second or third residue from the oligonucleotide of interest, respectively. ) T—OH (hereinafter referred to as “deleted body”) was measured by MS analysis.
• Example 19 Synthesis of HO- MeC (O)A(O)G(O)T(S)T-SUC-TOB using compound (Ib-1) as oxidant HO-T(S)T-SUC-TOB (250 mg, 88 ⁇ mol), dehydrated dichloromethane (4.4 mL) and dehydrated acetonitrile (1.3 mL) were added to the flask to prepare a solution.
• dG-CE phosphoramidite 149 mg, 176 ⁇ mol
• 5-ethylthio-1H-tetrazole 23 mg, 0.18 mmol
• 2,2,2-Trifluoroethanol 38 ⁇ L, 0.53 mmol was then added to the solution and the solution was stirred at room temperature for 30 minutes. Subsequently, water (70 ⁇ L) and compound (Ib-1) (59 mg, 18 mmol) were added and the solution was stirred at room temperature for 60 minutes. After sampling a portion of the solution, 2,3-dimethylfuran (46 ⁇ L, 0.88 mmol) and trifluoroacetic acid (102 ⁇ L, 1.32 mmol) were added sequentially to the solution, and the solution was stirred at room temperature for 80 minutes.
• Example 20 Synthesis of HO-MeC(O)A(O)G(O)T(S)T-SUC-TOB using compound (Ib-1) as oxidant ( with addition of aromatic amine)
• compound (Ib-1) as oxidant (with addition of aromatic amine)
• 2,6-xylidine 3 mol per 1 mol of compound (Ib-1) was added as an aromatic amine.
• SUC-TOB was obtained (121 mg).
• Example 21 Oligonucleotides that are 20-mers (DMTrO-Um(O)Cm(O)Am(O)Am(O)Gm(S)Gm(S)Am(S)Am(S)Gm(S) Am(S)Um(S)Gm(S)Gm(S)Cm(S)Am(S)Um(S)Um(S)Cm(S)Um-OH) (SEQ ID NO: 1) Solid Phase Synthesis Oligonucleotides that are 20-mer mers of the above sequence were synthesized by solid phase synthesis by the phosphoramidite method (in the above sequence, Am represents a 2′-O-methyladenosine residue, Cm represents a 2′ -O- represents a methylcytidine residue, Gm represents a 2'-O-methylguanosine residue, Um represents a 2'-O-methyluridine residue, (O) represents the formula (P3- 1) represents a phosphate este
• 5′-O-(4,4′-dimethoxytrityl)-2′-O-methyluridine supported at 41 ⁇ mol/g on a porous glass (CPG) solid phase carrier (manufactured by ChemGenes) (solid Amount of phase carrier: 24 mg, amount of 5′-O-(4,4′-dimethoxytrityl)-2′-O-methyluridine: 1 ⁇ mol) was placed in a column equipped with a glass filter, and an oligonucleotide synthesizer (trade name “ nS-8", manufactured by Gene Design), synthesis was carried out from the 3' side to the 5' side.
• CPG porous glass
• a mixed solution of 10% by weight acetic anhydride in THF and a THF solution of 1-methylimidazole and pyridine was used as a capping reagent, and a dichloromethane solution of trichloroacetic acid (TCA) was used as a detritylation reagent.
• TCA trichloroacetic acid
• an oxidizing agent (1) 0.01 M pyridine solution of compound (Ib-1), (2) 0.1 M pyridine solution of compound (Ic-1), or (3) 0.02 M compound (Ic- The acetonitrile solution of 1) was used.
• Dehydrated DMF (10 mL) was added to the solid to prepare a diluted solution.
• dehydrated DMF (2 mL) sodium hydroxide (0.48 g, 12 mmol), and tert-butyl mercaptan (1.4 mL, 12 mmol) were added to a 100 mL flask and stirred under ice-cooling to obtain After the diluent was added dropwise to the resulting mixture, the mixture was warmed to room temperature and stirred for 4 hours. Water (120 mL) was then slowly added dropwise to the mixture under an ice bath.
• the precipitated solid was collected by filtration, ethanol (400 mL) was added, the mixture was stirred at 100° C., and then the mixture was cooled to room temperature. The solid that precipitated out of the mixture was collected by filtration and the solid obtained was dried under reduced pressure (1.7 g). Also, the filtrate was concentrated under reduced pressure, ethanol (400 mL) was added, the mixture was stirred at 100° C., water (200 mL) was added dropwise, and the mixture was cooled to room temperature. The solid that precipitated out of the mixture was collected by filtration and the resulting solid was dried under reduced pressure (0.85g). The resulting 1.7 g of solid was mixed with 0.85 g of solid, of which 2.0 g was used in the next step.
• Synthesis Example 10 Synthesis of 2-phenyl-5-trifluoromethylisothiazolo[5,4-b]pyridin-3(2H)-one (Compound (Ic-12)) Into a 200 mL flask was added 2-chloro-5- (Trifluoromethyl)pyridine-3-carboxylic acid (2.5 g, 11 mmol) and thionyl chloride (37 mL) were added and the mixture was stirred at 80° C. for 4 hours. After thionyl chloride was distilled off under reduced pressure, toluene (30 mL) was added, and the resulting solution was concentrated under reduced pressure to obtain a concentrate.
• 2-chloro-5- (Trifluoromethyl)pyridine-3-carboxylic acid 2.5 g, 11 mmol
• thionyl chloride 37 mL
• Dehydrated DMF (5 mL) was added to the solid to prepare a diluted solution. Further, under an argon atmosphere, dehydrated DMF (2 mL), sodium hydroxide (0.47 g, 12 mmol) and tert-butyl mercaptan (1.3 mL, 11 mmol) were added to a 100 mL flask and stirred under ice cooling to obtain After the diluent was added dropwise to the resulting mixture, the mixture was warmed to room temperature and stirred for 4 hours. After that, water (70 mL) was slowly added dropwise to the mixture under ice-cooling.
• the precipitated solid was collected by filtration, ethanol (30 mL) was added, the mixture was stirred at 100° C., water (30 mL) was added dropwise, and the mixture was cooled to room temperature. The precipitated solid was collected by filtration, and the resulting solid was dried under reduced pressure. Ethanol (30 mL) was added to the resulting solid, the mixture was stirred at 100° C., water (30 mL) was added dropwise, and the mixture was cooled to room temperature. The precipitated solid was collected by filtration and the solid obtained was dried under reduced pressure (3.4 g).
• Synthesis Example 11 Synthesis of 2-phenyl-2H-1,2-benzothiazin-3(4H)-one (Compound (Ic-13))
• 2-chloropyridine-3-acetonitrile 5 .0 g, 33 mmol
• 4 mol/L aqueous sodium hydroxide solution 75 mL
• 6 mol/L hydrochloric acid 60 mL was slowly added dropwise under ice-cooling, and the mixture was stirred for 1 hour.
• the precipitated solid was collected by filtration, washed with 2-propanol (40 mL) and dried under reduced pressure (5.6 g).
• Dehydrated DMF (20 mL) was added to the concentrate to prepare a diluted solution. Also, under an argon atmosphere, dehydrated DMF (2 mL), sodium hydroxide (2.6 g, 64 mmol), and tert-butyl mercaptan (4.4 mL, 39 mmol) were added to a 300 mL flask and stirred for 10 minutes under ice-cooling. After the diluent was added dropwise to the obtained mixture, the mixture was stirred at 110° C. for 23 hours. After that, water (260 mL) was slowly added dropwise to the mixture under ice-cooling.
• Synthesis Example 12 Synthesis of 2-(4-methyl-2-pyridinyl)isothiazolo[5,4-b]pyridin-3(2H)-one (Compound (Ic-15)) -Chloropyridine-3-carbonyl chloride (5.0 g, 28 mmol) and anhydrous THF (12 mL) were added and the mixture was stirred under ice cooling. To the resulting mixture, a solution of triethylamine (4.8 mL, 34 mmol), 2-amino-5-methylpyridine (3.7 g, 34 mmol) and anhydrous THF (28 mL) was added dropwise over 30 minutes and warmed to room temperature. and stirred for 23 hours.
• Triethylamine 4.8 mL, 34 mmol
• 2-amino-5-methylpyridine 3.7 g, 34 mmol
• anhydrous THF 28 mL
• Dehydrated DMF (20 mL) was added to the concentrate to prepare a diluted solution. Further, under an argon atmosphere, dehydrated DMF (4 mL), sodium hydroxide (1.3 g, 34 mmol), and tert-butyl mercaptan (3.6 mL, 32 mmol) were added to a 300 mL flask and stirred under ice cooling to obtain After the diluent was added dropwise to the resulting mixture, the mixture was warmed to room temperature and stirred for 5 hours. After that, water (240 mL) was slowly added dropwise to the mixture over 25 minutes under ice-cooling.
• the precipitated solid was collected by filtration, ethanol (60 mL) was added, the mixture was stirred at 100° C., water (60 mL) was added dropwise, and the mixture was cooled under ice cooling. The precipitated solid was collected by filtration and the solid obtained was dried under reduced pressure (5.4 g).
• Synthesis Example 13 Synthesis of Oligonucleotide Precursor Having Phosphite Ester Bond Under an argon atmosphere, HO- Me C-SUC-TOB (15.1 g, 11.2 mmol) and Piv-TOB (30.1 g) were placed in a 500 mL flask. , 30.2 mmol), dehydrated chloroform (562 mL) and dehydrated acetonitrile (169 mL) were added to prepare a solution. Molecular sieves 3A (11.2 g), dG-CE phosphoramidite (18.9 g, 22.5 mmol) and triphenylphosphine (1.5 g, 5.6 mmol) were then added and the solution was stirred for 30 minutes.
• Examples 22-41 and Comparative Example 4 A mixture of DMTrO-G(III)A(S)G(S) Me C-SUC-TOB and Piv-TOB (100 mg: DMTrO- G(III)A(S)G(S) Me 2 C-SUC-TOB (52.8 mg, 0.018 mmol) and dry dichloromethane (0.9 mL) were added to prepare a solution. After adding pyridine (0.11 mmol) and water (8.8 ⁇ L) to the solution, various oxidizing agents (53 ⁇ mol) shown in Tables 7 to 9 below were added and stirred at room temperature for 3 hours for oxidation.
• the oligonucleotide DMTrO-G(O)A(S)G(S) Me C-SUC-TOB was synthesized.
• Triphenylphosphine (9.2 mg, 35 ⁇ mol) was then added and the solution was stirred at room temperature for 30 minutes. After that, acetonitrile (129 mL) was added, and the precipitated solid was collected by suction filtration using a Kiriyama funnel and then dried under reduced pressure. A mixture of S)G(S) MeC -SUC-TOB was obtained. m/z: Calcd. 3013.49, Found 3014.54 [M+H] +
• Compounds (Ib-2) to (Ib-5) and compound (Ib-8) used were those obtained in Synthesis Examples 3 to 7 described above.
• Compounds (Ic-11) to (Ic-13) and compound (Ic-15) used were those obtained in Synthesis Examples 9 to 12 described above.
• Compound (Ic-1), Compound (Ic-2), Compound (Ic-5), Compound (Ic-10), Compound (Ic-14), Compound (Ic-16) and Compound (Ic-17) are The one synthesized by the method described in Supporting Information of Non-Patent Document 2 was used. Other oxidizing agents used were commercially available products.
• sulfide Sulfurizing G(III) A(S) G(S) Me C-SUC-TOB to form DMTrO-G(S) A(S) G(S) Me C-SUC-TOB (hereinafter "sulfide" ) was synthesized to prepare a test solution containing an oxidant and a sulfide.
• oligonucleotides by suppressing the production of the above-mentioned truncated forms or desulfurized forms compared to the case where iodine is used.
• Oligonucleotides obtained by the production method of the present invention can be used for human or veterinary pharmaceuticals (RNA, DNA, oligonucleotide medicines, etc.), functional foods, foods for specified health uses, foods, chemical products, biological and industrial products. It can be used for various applications such as polymer materials.

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