Technical field
[0001]
The present invention relates to a method for manufacturing an oligonucleotide, and the nucleoside can be effectively used in the manufacturing method, a nucleotide or oligonucleotide.
BACKGROUND
[0002]
As a method for producing an oligonucleotide, currently, the solid phase method using the phosphoramidite method is widely used (Non-patent Document 1). Solid phase method is automated process optimization is performed also progressing, but it is advantageous in speed plane, there is a disadvantage that equipment constraints on the scale-up limitations. Also, have been studied the manufacturing method of oligonucleotides by a liquid phase method, liquid phase method, the operation is complicated, since lower yields, is difficult to synthesize a large amount and quickly multiple degree of polymerization of the oligonucleotide there is a disadvantage that is.
[0003]
　Recently, as an attempt to overcome the respective disadvantages of solid-phase method and liquid phase method, a manufacturing method using a hydrophobic group linked nucleosides (Patent Document 1), or 3 'hydroxyl groups are protected with certain organic groups nucleosides or manufacturing method using the oligonucleotides (Patent Document 2) discloses a.
[0004]
　Patent Document 2 described above, the following steps (i) ~
　(iv): (i) a nucleoside or oligonucleotide 5 'position hydroxyl-protecting groups (e.g., dimethoxytrityl group) removal
　(deprotection), (ii ) and nucleoside or oligonucleotide obtained in deprotected 3'-position hydroxy group is phosphoramidite of, and 5 'position hydroxyl condensation of protected nucleoside or oligonucleotide,
　phosphorylase obtained in (iii) condensation oxidation or sulfurization of the phosphite triester, and
　(iv) obtained in the oxidation or sulfurization, solid-liquid separation of the 5 'position hydroxyl group is protected oligonucleotide
production method of an oligonucleotide comprising is described. By repeating the above steps (i) ~ (iv), it is possible to extend the oligonucleotide chain.
CITATION
Patent Document
[0005]
Patent Document 1: JP 2010-275254 Patent Publication
Patent Document 2: WO 2012/157723
Non-patent literature
[0006]
非特許文献1 : S. L. Beaucage, D. E. Bergstorm, G. D. Glick, R. A. Jones, Current Protocols in Nucleic Acid Chemistry; John Wiley & Sons (2000)
Summary of the Invention
Problems that the Invention is to Solve
[0007]
　An object of the present invention is to provide a manufacturing method of an oligonucleotide capable of performing condensation efficiently.
Means for Solving the Problems
[0008]
　The present inventors have conducted intensive studies, to change the process sequence described in Patent Document 2,
　at least one of the groups such as (I) 5 'position hydroxyl group is not protected, and 3' hydroxyl group nucleosides protected with a protective group, a nucleotide or oligonucleotide (a) or the like, 3 'hydroxyl group or 3' amino group is phosphoramidite of, and 5 'position nucleoside hydroxyl group is protected, or oligonucleotide (b) condensation of,
　(II) obtained phosphite triester condensation oxide or sulfide (c),
　the protection of the 5'-position hydroxyl group of (III) obtained oligonucleotide oxide or sulfide (d) removal of group (deprotection), and
　(IV) obtained in the deprotection, purification by solid-liquid separation or extraction of the 5'-position hydroxyl group is not protected oligonucleotides (e)
performing By been found that may perform efficiently condensed.
[0009]
　The repetition of steps described in Patent Document 2 (i) ~ (iv), deprotection (step (i)) condensation after (step (ii)) but is carried out, the above-described step (I) ~ (IV) in the repetition, the solid-liquid separation or extraction condensation after (step (IV)) (step (I)) is performed. Therefore, in the manufacturing method comprising the above steps (I) ~ (IV), may be impurities arising in the deprotection is not present at the time of condensation, for efficient condensation. Although the present invention based on the findings, such as is as follows.
[0010]
　[1] The following step (1), (3), (4) and (6) an oligonucleotide of the manufacturing method
　comprising: (1) the non-polar solvent,
　5 'hydroxyl group is not protected, nucleobase amino group and imino group, 2 'position hydroxyl, 3' of the ribose residue position hydroxyl group and 3'-amino group, and 3 of the deoxyribose residues' at least one group selected from the position hydroxyl group and 3'-amino group but not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and optionally a nucleoside even though other groups are further protected with a protecting group to be used in nucleic acid synthesis, or oligonucleotide (a), or
　5 'position hydroxyl group is not protected, 3' one hydroxyl group is -OL the position phosphoric acid group n1 (wherein, L -OH n1 represents an organic group.) is replaced, -OL N1 - OH of the hydroxyl groups are not eliminated under acidic conditions, are protected with a removable protecting group under basic conditions, and other groups are further protected may be substituted nucleotides or oligonucleotides with a protecting group to be used in nucleic acid synthesis nucleotide (alpha)
to the reaction solution containing,
　3 'hydroxyl group or 3' amino group is phosphoramidite of 5'-position hydroxyl group is protected with a temporary protecting group which can be removed under acidic conditions, and other groups There further is not removed under acidic conditions, may nucleosides be protected by a protecting group selected from the protecting groups used in the removable protecting group and nucleic acid synthesis under basic conditions, or oligonucleotide (b)
Was added and the nucleoside, nucleotide or oligonucleotide (a) or a substituted or oligonucleotide (alpha) and a nucleoside, nucleotide or oligonucleotide (b) and condensing, 5 'hydroxyl group which can be removed under acidic conditions obtaining a reaction solution containing the protected phosphite triester temporary protecting group (c);
　(3) by adding an oxidizing agent or a sulfurizing agent to a reaction solution containing phosphite triester body (c), phosphite oxidized or sulfurized triester body (c), the step 5 'hydroxyl group to obtain a reaction liquid containing a protected oligonucleotide with a temporary protecting group which can be removed under acidic conditions (d);
　(4) oxidizing or by adding an acid to the reaction solution after sulfurization 5 'position is removed a temporary protecting group for a hydroxyl group, 5' oligonucleotide-position hydroxyl group is not protected (E) obtaining a reaction solution containing; and
　(6) adding a polar solvent to a reaction solution containing oligonucleotides (e), purifying the solid-liquid separation or extraction of the oligonucleotides (e).
[0011]
　[2] in step (1),
　5 'hydroxyl group is not protected, an amino group and imino group nucleobases, 2 of ribose residue' position hydroxyl group and 3'-position a hydroxyl group, and 3 of the deoxyribose residues at least one group selected from the 'position hydroxyl group is not removed under acidic conditions, it is protected with a removable protecting group under basic conditions, and other groups are further protected with a protecting group to be used in nucleic acid synthesis which may be a nucleoside, nucleotide or oligonucleotide (a), or
　5 'position hydroxyl group is not protected, the 3'-position a hydroxyl group of phosphate group -OL n1 in -OH (wherein, L n1 of the organic and replaced with a group.), -OL n1 hydroxyl group of -OH is not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and use the other group is more nucleic acid synthesis need Protecting group in a protected or may be substituted nucleotides or oligonucleotides (alpha), and
　3'-position a hydroxyl group is phosphoramidite of 5'-position hydroxyl group is protected with a removable temporary protecting group under acidic conditions, and other groups are further not removed under acidic conditions, may nucleosides be protected by a protecting group selected from the protecting groups used in the removable protecting group and nucleic acid synthesis under basic conditions, nucleotide or oligonucleotide nucleotides (b)
the production method according to [1] to use.
[0012]
　[3] In step (1),
　5 'hydroxyl group is not protected, an amino group and imino group nucleobases, 2 of ribose residue' position hydroxyl group and 3'-position a hydroxyl group, and 3 of the deoxyribose residues at least one group selected from the 'position hydroxyl group is not removed under acidic conditions, it is protected with a removable protecting group under basic conditions, and other groups are further protected with a protecting group to be used in nucleic acid synthesis which may be a nucleoside or oligonucleotide (a), and
　3 'position hydroxyl group is phosphoramidite of 5'-position hydroxyl group is protected with a removable temporary protecting group under acidic conditions, and other groups are more nucleic acid protected with a protecting group used in the synthesis which may be a nucleoside or oligonucleotide (b)
the production method according to [1] to use.
[0013]
　[4] Step (1) prior to the following and step (3) of the further steps of (2) the containing [1] to [3] The method according to any
　one: (2) condensation adding a quenching agent to the reaction solution after.
　[5] quenching agent used in step (2) The production method according to the alcohol is at least one selected from phenols and amines [4].
[0014]
　[6] to the reaction solution before the step (1) and the step (4) after the mixture of carboxylic acids and organic bases, adding an inorganic acid or an amine, in the step (3), as a sulfurizing agent 5- [ (N, N-dimethylaminomethylidene) the method according to any one of [1] to [5] for adding amino]-3H-1,2,4-dithiazole-3-thione in the reaction solution .
　[7] The amount of the basic nitrogen atoms of the organic base has the production method according to [6] is 1 to 2 moles with respect to the carboxy group 1 mol having a carboxylic acid.
[0015]
　[8] sulfurizing agent used in step (3) is, 3H-1,2-benzodithiol-3-one 1,1-dioxide, 3H-1,2-benzodithiole-3-one, phenylacetyl disulfide, least one selected tetraethyl thiuram disulfide, dipentamethylenethiuram tetrasulfide, phenyl-3H-1,2,4-dithiazole-3-one, from 3-amino-1,2,4-dithiazole-5-thione and sulfur the process according to any one of [1] to [5] is.
　[9] The oxidizing agent used in step (3) is iodine, (1S) - (+) - (10- camphorsulfonic sulfonyl) oxaziridine, tert- butyl hydroperoxide, 2-butanone peroxide, 1,1-dihydroperoxy cyclododecane, bis (trimethylsilyl) peroxide and at least one a is the selected from m- chloroperbenzoic acid [1] to process according to any one of [5].
[0016]
　[10] temporary custody group, The process according to any one of the a dimethoxytrityl group or monomethoxytrityl group [1] to [9].
　[11] Non-polar solvent, prepared as described in any one of halogenated solvents, aromatic solvents, said at least one selected from ester solvents and aliphatic solvents [1] to [10] Method.
　[12] step acid used in (4), trifluoroacetic acid, dichloroacetic acid, trifluoromethanesulfonic acid, trichloroacetic acid, methanesulfonic acid, hydrochloric acid, at least one a is the selected from acetic acid and p- toluenesulfonic acid [1] - the process according to any one of [11].
　[13] not removed under acidic conditions, a protective group removable under basic conditions, aliphatic having or having a linear aliphatic hydrocarbon group having 10 or more carbon atoms, or branched one or more of the process according to any one of [1] to [12] having a hydrocarbon group having at least one, and an organic group has a total carbon number of 14 to 300.
[0017]
　[14] Step (6), and adding a polar solvent to a reaction solution containing oligonucleotides (e), any of the a step of purifying the solid-liquid separation of oligonucleotides (e) [1] ~ [13] the method according to one or.
　[15] Step polar solvent used in (6) The production method according to any one of the nitrile-based solvent [1] to [14].
[0018]
　[16] Step (4) before after and step (6) of the further steps of (5) above containing [1] - [15] The method according to any
　one: (5) Oligo a step of neutralization by adding a base to the reaction solution containing a nucleotide (e).
　[17] The base used in step (5) is, pyridine, 2,4,6-trimethylpyridine, benzimidazole, 1,2,4-triazole, N- phenylimidazole, 2-amino-4,6-dimethylpyrimidine , 1,10-phenanthroline, selected imidazole, N- methylimidazole, 2-chloro-benzimidazole, 2-bromo-benzimidazole, 2-methylimidazole, 2-phenyl-benzimidazole, from N- phenyl benzimidazole and 5-nitro-benzimidazole the process according to at least one a is the [16] to be.
[0019]
　[18] after the step (6), further the following steps (7) said containing [1] - [17] The method according to any one:
　The protecting group of the resulting oligonucleotide (7) after all removed, isolating the oligonucleotide unprotected.
[0020]
　[19] Step (4) of the temporary removal of the protecting group or carried out in the presence of a cation scavenger, or the addition of a cation scavenger to the reaction solution after removal reaction of the temporary protecting groups [1] to [18] the process according to any one of.
　[20] cation scavenger, pyrrole, 2-methylpyrrole, 3-methylpyrrole, 2,3-dimethylpyrrole, 2,4-dimethylpyrrole, indole, 3-methylindole, 4-methylindole, 5-methylindole , 6-methylindole, 7-methylindole, 5,6-dimethyl indole, 6,7-dimethyl indole, 2-methylfuran, 2,3-dimethyl furan, 2-methyl-3- (methylthio) furan, and placements the process according to at least one a is the selected from furan [19].
[0021]
　[21] The following steps (1 '), (3'), (4 ') and (6') oligonucleotides manufacturing method
　comprising: (1 ') a non-polar solvent,
　3' hydroxyl group or 3 ' no amino group is protected, an amino group and imino group nucleobases, 2'-position hydroxyl group and the 5'-position hydroxyl group of the ribose residue, and at least one group selected from the 5'-position hydroxyl group of deoxyribose residues not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and optionally nucleosides be protected with a protecting group other groups are further used in nucleic acid synthesis, or oligonucleotides ( a '), or
　3' position hydroxyl group or 3 'amino group is not protected, 5' one hydroxyl group in position phosphate group -OL n1 in -OH (wherein, L n1 represents an organic group. ) it has been replaced by, - L n1 hydroxyl group of -OH is not removed under acidic conditions, are protected with a removable protecting group under basic conditions, it may be and the other groups are further protected with a protecting group to be used in nucleic acid synthesis substitution 'or oligonucleotide (alpha)
to the reaction solution containing,
　5' hydroxyl group is phosphoramidite of, 3 'hydroxyl group or 3' amino group is protected with a temporary protecting group which can be removed under acidic conditions, and other groups are further not removed under acidic conditions, may nucleosides be protected by a protecting group selected from the protecting groups used in the removable protecting group and nucleic acid synthesis under basic conditions, nucleotide or oligonucleotide nucleotide (b ')
Was added and the nucleoside, nucleotide or oligonucleotide (a ') or a substituted or oligonucleotide (alpha') and a nucleoside, nucleotide or oligonucleotide (b ') and the condensation of the 3' position hydroxyl group or 3 'amino ; 'groups obtaining a reaction liquid containing a phosphite triester protected with a removable temporary protective group under acidic conditions (c)
　a reaction solution containing (phosphite triester body (c') 3) ' by adding an oxidizing agent or a sulfurizing agent, phosphite triester (c ') the oxide or sulfide, 3' are protected at the a position hydroxyl group or 3 'amino group temporary protecting group which can be removed under acidic conditions to oligonucleotide 'to obtain a reaction liquid containing a step; (d)
　(4') 3 by adding an acid to the reaction mixture after oxidation or sulfurization 'position hydroxyl group or 3' temporary amino group The protecting group is removed, 3 'hydroxyl group or 3' amino groups oligonucleotides unprotected (e ') to obtain a reaction solution containing; and
　(6') reaction containing oligonucleotides (e ') step by adding a polar solvent, is purified by solid-liquid separation or extraction of the oligonucleotides (e ') to.
[0022]
　[22] 'In step (1)
　3' position hydroxyl group is not protected, an amino group and imino group nucleobases, 2'-position hydroxyl group and the 5'-position hydroxyl group of the ribose residue, as well as the deoxyribose residues 5 'position of at least one group selected from hydroxyl group, not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and protected with a protecting group other groups are further used in nucleic acid synthesis which may be a nucleoside, nucleotide or oligonucleotide (a '), or
　3'-position hydroxy group is not protected, one of the hydroxyl groups of 5' position phosphate group -OL n1 in -OH (wherein, L n1 represents an organic group. are replaced), -OL n1 hydroxyl group of -OH is not removed under acidic conditions, it is protected with a removable protecting group under basic conditions, and other groups more nucleic acid synthesis use to Protecting groups in a protected substituted may be a nucleotide or oligonucleotide to be (alpha '), as well as
　5' hydroxyl group is phosphoramidite of, 3'-position hydroxy group is protected with a removable temporary protective group under acidic conditions and other groups are further not removed under acidic conditions, may nucleosides be protected by a protecting group selected from the protecting groups used in the removable protecting group and nucleic acid synthesis under basic conditions, nucleotides or oligonucleotides (b ')
production method according to [21] for use.
[0023]
　[23] 'In step (1)
　3' position hydroxyl group is not protected, one of the hydroxyl groups of 5 'position phosphate group -OL n1 -OH (wherein, L n1 represents an organic group.) and replaced, -OL n1 hydroxyl group of -OH is not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and protected with a protecting group other groups are further used in nucleic acid synthesis also substituent or oligonucleotide have (alpha '), and the
　5' position hydroxyl group is phosphoramidite of, 3'-position hydroxy group is protected with a temporary protecting group which can be removed under acidic conditions, and other groups There further is not removed under acidic conditions, basic conditions with a removable protecting group and optionally nucleosides be protected with a protecting group selected from the protecting groups used for nucleic acid synthesis or oligonucleotide (b ')
a use The method according to [21] for use.
[0024]
　[24] 'In step (1)
　3' position hydroxyl group is not protected, an amino group and imino group nucleobases, 2'-position hydroxyl group and the 5'-position hydroxyl group of the ribose residue, as well as the deoxyribose residues 5 'position of at least one group selected from hydroxyl group, not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and protected with a protecting group other groups are further used in nucleic acid synthesis which may be a nucleoside or oligonucleotide (a '), as well as
　5' hydroxyl group is phosphoramidite of, 3'-position hydroxy group is protected with a removable temporary protective group under acidic conditions, is and other groups Furthermore protected also good nucleoside or oligonucleotide with a protecting group to be used in nucleic acid synthesis (b ')
production method according to [21] for use.
[0025]
　[25] Step (1 ') before after and step (3'), the following additional steps (2 ') said containing [21] - [24] a method of manufacturing according to any one
　of: ( adding a quenching agent to the 2 ') reaction solution after the condensation.
　[26] quenching agent used in step (2 ') The production method according to the alcohol is at least one selected from phenols and amines [25].
[0026]
　[27] To the reaction solution before the step (1 ') after and the step (4'), mixtures of carboxylic acids and organic bases, adding an inorganic acid or an amine, in the step (3 '), as a sulfurizing agent 5 - [(N, N- dimethylaminomethylidene) amino]-3H-1,2,4-dithiazole-3-thione according to any one of [21] - [26] to be added to the reaction solution the method of production.
　[28] The amount of the basic nitrogen atoms of the organic base has the production method according to [27] is 1 to 2 moles with respect to the carboxy group 1 mol having a carboxylic acid.
[0027]
　[29] sulfurizing agent used in the step (3 ') is, 3H-1,2-benzodithiol-3-one 1,1-dioxide, 3H-1,2-benzodithiole-3-one, phenylacetyl disulfide , tetraethyl thiuram disulfide, dipentamethylenethiuram tetrasulfide, phenyl-3H-1,2,4-dithiazole-3-one, at least a selected from 3-amino-1,2,4-dithiazole-5-thione and sulfur the process according to any one of [21] - [26] one in which.
　[30] The oxidizing agent used in the step (3 ') is iodine, (1S) - (+) - (10- camphorsulfonic sulfonyl) oxaziridine, tert- butyl hydroperoxide, 2-butanone peroxide, 1,1-dihydro peroxy cyclododecane bis process according to any one of the at least one selected from (trimethylsilyl) peroxide and m- chloroperbenzoic acid [21] - [26].
[0028]
　[31] temporary custody group hydroxyl groups, the manufacturing method according to any one of the a dimethoxytrityl group or monomethoxytrityl group [21] - [30].
　[32] Non-polar solvent, prepared as described in any one of halogenated solvents, aromatic solvents, said at least one selected from ester solvents and aliphatic solvents [21] - [31] Method.
　[33] the acid used in step (4 '), trifluoroacetic acid, dichloroacetic acid, trifluoromethanesulfonic acid, trichloroacetic acid, methanesulfonic acid, hydrochloric acid, is at least one selected from acetic acid and p- toluenesulfonic acid the [21] - the process according to any one of [32].
　[34] not removed under acidic conditions, a protective group removable under basic conditions, aliphatic having or having a linear aliphatic hydrocarbon group having 10 or more carbon atoms, or branched one or more of the process according to any one of [21] - [33] having a hydrocarbon group having at least one, and an organic group has a total carbon number of 14 to 300.
[0029]
　[35] Step (6 ') is, oligonucleotides (e' adding a polar solvent to a reaction solution containing a) said a step of purifying the solid-liquid separation of oligonucleotides (e ') [21] ~ [34 the process according to any one of.
　[36] Step polar solvent used in (6 ') The production method according to any one of the nitrile-based solvent [21] - [35].
[0030]
　[37] Step (4 ') before after and step (6'), further the following steps (5 ') said containing [21] - [36] a method of manufacturing according to any one
　of: ( 5 ') oligonucleotides (e') a step of neutralizing by adding base to the reaction solution containing.
　[38] The base used in step (5 ') is, pyridine, 2,4,6-trimethylpyridine, benzimidazole, 1,2,4-triazole, N- phenylimidazole, 2-amino-4,6-dimethyl pyrimidine, 1,10-phenanthroline, imidazole, N- methylimidazole, 2-chloro-benzimidazole, 2-bromo-benzimidazole, 2-methylimidazole, 2-phenyl-benzimidazole, from N- phenyl benzimidazole and 5-nitro-benzimidazole the process according to at least one a is the [37] to be selected.
[0031]
　[39] 'after further steps of (7 step (6)') said containing [21] - [38] a method of manufacturing according to any one
　of: (7 ') resulting oligonucleotide after all deprotection, isolating the oligonucleotide unprotected.
[0032]
　[40] Step (4 ') of the temporary removal of the protecting group or carried out in the presence of a cation scavenger, or the addition of a cation scavenger to the reaction solution after removal reaction of the temporary protecting group [21] - [39 the process according to any one of.
　[41] cation scavenger, pyrrole, 2-methylpyrrole, 3-methylpyrrole, 2,3-dimethylpyrrole, 2,4-dimethylpyrrole, indole, 3-methylindole, 4-methylindole, 5-methylindole , 6-methylindole, 7-methylindole, 5,6-dimethyl indole, 6,7-dimethyl indole, 2-methylfuran, 2,3-dimethyl furan, 2-methyl-3- (methylthio) furan, and placements the process according to at least one a is the selected from furan [40].
[0033]
　[42] formula (a-II):
[0034]
[Formula 1]

[0035]
Wherein,
　m represents an integer of 0 or more;
　m number of Base are each independently be protected indicates nucleic acid base which may;
　m + 1 pieces of X are each independently a hydrogen atom, halogen atom, optionally protected hydroxyl group or 2-position represents a divalent organic group bonded to a carbon atom and 4 carbon atoms,;
　m pieces of R 10 are each independently an oxygen atom or a sulfur atom shown;
　m-number of R p1 are each independently a protecting group of a phosphate group;
　L represents a single bond or the formula (a1) or (a1 '):
[0036]
[Formula 2]

[0037]
(Wherein,
　* indicates the bonding position to
　Y; ** indicates the bonding position to the oxygen
　atom; R 1 and R 2 are each independently, C 1-22 represents a hydrocarbon group;
　L 1 is C divalent optionally substituted 1-22 represents a hydrocarbon
　group; L 2 is either a single bond, or C *** (= O) N (R 3 ) -R 4 -N (R 5 ) **** (wherein, *** is L 1 represents a bonding position with, **** denotes the bonding position to C = O, R 4 is C 1 -22 represents an alkylene group, R 3 and R 5 are each independently a hydrogen atom or a C 1-22 represents an alkyl group, or R 3 and R 5 are taken together, may form a ring. ) A group represented by. A group represented by);
　Y is a single bond, an oxygen atom or NR (R, represents a hydrogen atom, an alkyl group or an aralkyl group); the. And
　Z has the formula (a2), the formula ( a2 ') or formula (a2 "):
[0038]
[Formula 3]

[0039]
Wherein
　* indicates the binding position;
　R 6 is a hydrogen atom, or R b is, when a group represented by the following formula (a3), the ring A or ring B R 6 is R 8 represents a single bond or -O- together with ring a or ring B and ring C may form a fused ring with;
　k is an integer of 1 to 4;
　the k Q are each independently, -O -, - C (= O) -, - C (= O) O -, - OC (= O) -, - C (= O) NH- or - NHC (= O) - are shown;
　k-number of R 7 each independently represent a hydrocarbon group is a linear aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker or branched chain having at least one aliphatic hydrocarbon group having 1 or more, and the total number of carbon atoms represents an organic group is 14 or more 300 or less;
　ring a Contact And ring B are each independently, k-number of QR 7 in addition to further halogen atom, a C substituted with a halogen atom 1-6 optionally substituted alkyl group, and a halogen atom C 1-6 may have a substituent group selected from the group consisting of an alkoxy group;
　R a represents a hydrogen atom; and
　R b is a hydrogen atom or the formula, (a3):
[0040]
[Chemical Formula 4]

[0041]
(Wherein,
　* indicates the bonding position;
　j is 0 to an integer of 4;
　j-number of Q are each independently a as defined above;
　j-number of R 9 is each independently Te, at least one organic aliphatic hydrocarbon radical having either a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker, or branched one or more of and, and the total number of carbon atoms is an organic group of 14 to 300;
　R 8 is either a hydrogen atom, or R of ring a or ring B 6 taken together with a single bond or -O- Te, ring a or ring B and ring C may form a fused ring with; and
　ring C, j-number of QR 9 in addition to further halogen atom, a C substituted with a halogen atom 1 -6 alkyl groups, and may be substituted by a halogen atom C 1- May have a substituent group selected from the group consisting of an alkoxy group.) In either a group represented, or
　R a and R b are taken together to form an oxo group. It shows a group represented by.
　R 11 represents a methyl group, R 12 and R 13 are each independently C 1-5 represents an alkyl group, or R 11 and R 12 in the together, the carbon and nitrogen atoms they are attached it may form a nitrogen-containing hydrocarbon ring of 5-membered or 6-membered together. ]
Nucleoside or oligonucleotide represented by.
　[43] R p1 is, -CH 2 CH 2 WG (wherein, WG represents an electron-withdrawing group.) Nucleoside or oligonucleotide according to the a group represented by [42].
　[44] R 11 is a methyl group, R 12 and R 13 are each C is independently 1-5 nucleoside or oligonucleotide according to the an alkyl group [42] or [43].
　[45] the m is 0 [42] - nucleosides according to any one of [44].
[0042]
　[46] Formula (a-IV):
[0043]
[Formula 5]

[0044]
Wherein,
　m represents an integer of 0 or more;
　m + 1 pieces of Base are each independently be protected indicates nucleic acid base which may;
　m + 1 pieces of X are each independently a hydrogen atom, halogen atom, optionally protected hydroxyl group or 2-position represents a divalent organic group bonded to a carbon atom and 4 carbon atoms,;
　m pieces of R 10 are each independently an oxygen atom or a sulfur atom shown;
　m-number of R p1 are each independently a protecting group of a phosphate group;
　L represents a single bond or the formula (a1) or (a1 '):
[0045]
[Formula 6]

[0046]
(Wherein,
　* indicates the bonding position to
　Y; ** indicates the bonding position to the oxygen
　bond; R 1 and R 2 are each independently, C 1-22 represents a hydrocarbon group;
　L 1 is C divalent optionally substituted 1-22 represents a hydrocarbon
　group; L 2 is either a single bond, or C *** (= O) N (R 3 ) -R 4 -N (R 5 ) **** (wherein, *** is L 1 represents a bonding position with, **** denotes the bonding position to C = O, R 4 is C 1 -22 represents an alkylene group, R 3 and R 5 are each independently a hydrogen atom or a C 1-22 represents an alkyl group, or R 3 and R 5 are taken together, may form a ring. ) A group represented by. Represented by a group in);
　. Y represents a single bond, an oxygen atom or NR, (R represents a represents a hydrogen atom, an alkyl group or an aralkyl group); and
　Z 'has the formula (a2 "):
[0047]
[Chemical Formula 7]

[0048]
Wherein
　* indicates the binding position;
　R 6 is a hydrogen atom, or R b is, when a group represented by the following formula (a3) is R 8 together with single bond or -O- and shows, may form a fused ring together with ring B and ring C;
　k is from 1 to an integer of 4;
　k number of Q are each independently, -O -, - C (= O) -, - C (= O) O -, - OC (= O) -, - C (= O) NH- or -NHC (= O) - indicates;
　k number of R 7 are each independently an aliphatic hydrocarbon having either a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker, or branched one or more having at least one group, and total carbon number represents an organic group is 14 or more 300 or less;
　ring B, k-number of QR 7 in addition to further halo Gen atom, a C substituted with halogen atom 1-6 alkyl group, and even a C substituted by a halogen atom 1-6 may have a substituent group selected from the group consisting of an alkoxy group
　well; R AIt represents a hydrogen atom; and
　R b is a hydrogen atom or the formula, (a3):
[0049]
[Formula 8]

[0050]
(Wherein,
　* indicates the bonding position;
　j is 0 to an integer of 4;
　j-number of Q are each independently a as defined above;
　j-number of R 9 is each independently Te, at least one organic aliphatic hydrocarbon radical having either a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker, or branched one or more of and, and the total number of carbon atoms is an organic group of 14 to 300;
　R 8 is either a hydrogen atom, or R 6 represents a single bond or -O- together with ring B and ring It may form a fused ring together with C; and
　ring C, j-number of QR 9 in addition to further halogen atom, a C substituted with halogen atom 1-6 alkyl group, and a halogen atom unsubstituted or substituted C have 1-6 or consisting alkoxy group Or a group represented by the selected may have a substituent.), Or
　R a and R b are taken together to form an oxo group. It shows a group represented by. ]
In represented nucleoside or oligonucleotide.
　[47] R p1 is, -CH 2 CH 2 WG (wherein, WG represents an electron-withdrawing group.) Nucleoside or oligonucleotide according to the a group represented by [46].
　[48] nucleosides according to the m is 0 [46] or [47].
[0051]
　[49] formula (I):
[0052]
[Formula 9]

[0053]
Wherein,
　m represents an integer of 0 or more;
　m + 1 pieces of Base are each independently be protected indicates nucleic acid base which may;
　m + 1 pieces of X are each independently a hydrogen atom, halogen atom, optionally protected hydroxyl group or 2-position represents a divalent organic group bonded to a carbon atom and 4 carbon atoms,;
　m pieces of R 10 are each independently an oxygen atom or a sulfur atom shown;
　m-number of R p1 are each independently a protecting group of a phosphate
　group; R n1 and R n2 one is a hydrogen atom, the remaining one has the formula (II):
[0054]
[Formula 10]

[0055]
{Wherein,
　* indicates the bonding
　position; R 10 represents an oxygen atom or a sulfur
　atom; R p1 is a protecting group of a phosphate
　group; L n1 represents an organic group;
　L is a single bond , or the formula (a1) or (a1 '):
[0056]
[Of 11]

[0057]
(Wherein,
　* indicates the bonding position to
　Y; ** indicates the bonding position to the oxygen
　atom; R 1 and R 2 are each independently, C 1-22 represents a hydrocarbon group;
　L 1 is C divalent optionally substituted 1-22 represents a hydrocarbon
　group; L 2 is either a single bond, or C *** (= O) N (R 3 ) -R 4 -N (R 5 ) **** (wherein, *** is L 1 represents a bonding position with, **** denotes the bonding position to C = O, R 4 is C 1 -22 represents an alkylene group, R 3 and R 5 are each independently a hydrogen atom or a C 1-22 represents an alkyl group, or R 3 and R 5 are taken together, may form a ring. ) A group represented by. A group represented by);
　Y is a single bond, an oxygen atom or NR (R, represents a hydrogen atom, an alkyl group or an aralkyl group); the. And
　Z has the formula (a2), the formula ( a2 ') or formula (a2 "):
[0058]
[Chem. 12]

[0059]
Wherein
　* indicates the binding position;
　R 6 is a hydrogen atom, or R b is, when a group represented by the following formula (a3), the ring A or ring B R 6 is R 8 represents a single bond or -O- together with ring a or ring B and ring C may form a fused ring with;
　k is an integer of 1 to 4;
　the k Q are each independently, -O -, - C (= O) -, - C (= O) O -, - OC (= O) -, - C (= O) NH- or - NHC (= O) - are shown;
　k-number of R 7 each independently represent a hydrocarbon group is a linear aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker or branched chain having at least one aliphatic hydrocarbon group having 1 or more, and the total number of carbon atoms represents an organic group is 14 or more 300 or less;
　ring a Contact And ring B are each independently, k-number of QR 7 in addition to further halogen atom, a C substituted with a halogen atom 1-6 optionally substituted alkyl group, and a halogen atom C 1-6 may have a substituent group selected from the group consisting of an alkoxy group;
　R a represents a hydrogen atom; and
　R b is a hydrogen atom or the formula, (a3):
[0060]
[Formula 13]

[0061]
(Wherein,
　* indicates the bonding position;
　j is 0 to an integer of 4;
　j-number of Q are each independently a as defined above;
　j-number of R 9 is each independently Te, at least one organic aliphatic hydrocarbon radical having either a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker, or branched one or more of and, and the total number of carbon atoms is an organic group of 14 to 300;
　R 8 is either a hydrogen atom, or R of ring a or ring B 6 taken together with a single bond or -O- Te, ring a or ring B and ring C may form a fused ring with; and
　ring C, j-number of QR 9 in addition to further halogen atom, a C substituted with a halogen atom 1 -6 alkyl groups, and may be substituted by a halogen atom C 1- May have a substituent group selected from the group consisting of an alkoxy group.) In either a group represented, or
　R a and R b are taken together to form an oxo group. It shows a group represented by. A group represented by}. ]
In represented by nucleotide or oligonucleotide.
　[50] R n1 is a group represented by the formula (II), R n2 or oligonucleotide according to the is a hydrogen atom [49].
　[51] L n1 nucleotide or oligonucleotide according to the an ethylene group [49] or [50].
　[52] the m is 0 [49] - nucleotides according to any one of [51].
Effect of the invention
[0062]
　According to the manufacturing method of the oligonucleotides of the present invention, it is possible to perform condensation efficiently.
DESCRIPTION OF THE INVENTION
[0063]
[Term]
　Unless defined otherwise, all technical and scientific terms used herein have the same meaning as the present invention is commonly understood by one of ordinary skill in the belonging art. Although any methods and materials similar or equivalent to those described herein, can be used in the practice or testing of the present invention, the preferred methods and materials are described below. All publications and patents mentioned herein, for example, are described in publications can be used in connection with the described invention, constructs and for the purpose of describing and disclosing the methodologies, hereby by reference It is incorporated in the book.
[0064]
　In the present specification, the a structural unit of the oligonucleotide "nucleoside" nucleobase sugar (e.g., 2-deoxyribose, ribose, 2-position carbon atom and 4 carbon atoms are linked by a divalent organic group and it means 2-deoxyribose or compounds linked by N- glycoside into 1-position of the ribose, etc.).
　"Sugar" herein, also encompasses ribose amino sugar hydroxyl groups are replaced with amino group, and the 2-position hydroxyl group is replaced by a halogen atom.
[0065]
　The 2-position 2-deoxyribose or ribose carbon atoms and 4-position carbon atom is bound by a divalent organic group, for example, include the following compounds.
[0066]
[Formula 14]

[0067]
　The amino sugars, for example, the 3-position hydroxyl group is replaced by a amino group of 2-deoxyribose, ribose position 3 hydroxyl groups are replaced with amino group, and 3-position hydroxyl group is an amino group, as shown below, 2-position hydroxyl group include ribose replaced by halogen (in the following formulas, X s is a halogen atom.).
[0068]
[Formula 15]

[0069]
　As used herein, "phosphate group" are, -O-P (O) (OH) 2 as well, based on the oxygen atom is replaced by a sulfur atom or NH (e.g., -O-P (S) ( OH ) 2 , -NH-P (O) (OH) 2 , -NH-P (S) (OH) 2 ) also encompasses. Also, a hydroxyl group (-OH) is -OR in phosphate group p (wherein, R p represents an organic group such as a protecting group of the phosphate group) group replaced with (e.g., protected phosphate group ) it is also included in the "phosphorus acid group".
[0070]
　As used herein, the term "nucleotide" means a compound which phosphate group is bonded to the nucleoside. The nucleotides 3 'hydroxyl group or the 5' position hydroxyl group is replaced with a phosphate group, for example, compounds can be mentioned are (the following formula represented by the following formula, R m1 and R m @ 2 are each independently hydrogen atom or the organic group (excluding nucleoside residues) indicates, X m represents a hydrogen atom, a hydroxyl group or a halogen atom.).
[0071]
[Chemical Formula 16]

[0072]
　As used herein, the term "oligonucleotide" refers to a compound which nucleotides are linked one or more nucleosides. Incidentally, the "oligonucleotide", phosphorothioate oligonucleotides oxygen atom of the phosphate group is replaced by a sulfur atom, oligonucleotides phosphate groups -O- is replaced with -NH-, hydroxyl groups in the phosphoric acid group (-OH) is -OR p (wherein, R p represents an organic group) are also encompassed oligonucleotides replaced. The number of nucleosides of the oligonucleotide is not particularly limited in the present invention, preferably 3 to 50, more preferably from 5 to 30.
[0073]
　Herein, 'the term "position amino group, nucleosides, 3 or oligonucleotide 3' 'refers to an amino group bound to a carbon atom of a.
　Herein, 'the term "position amino group, nucleosides, 5 nucleotides or oligonucleotide 5' 'refers to an amino group bound to a carbon atom of a.
[0074]
　Herein, 'the term "position phosphate group, 3 nucleotides or oligonucleotide 3' 'means a phosphate group bonded to a carbon atom of a.
　Herein, 'the term "position phosphate group, 5 nucleotides or oligonucleotide 5' 'means a phosphate group bonded to a carbon atom of a.
[0075]
　As used herein, the term "nucleobase" is not particularly limited as long as it is used in the synthesis of nucleic acid, for example, Shitoshiru group, uracil group, a pyrimidine base, adenyl group such thyminyl group, such as a guanyl group mention may be made of the purine base. In addition, the "optionally protected nucleobase", for example, means that the adenyl group is a nucleobase having an amino group, in guanyl group or Shitoshiru group, an amino group may be protected, amino groups of the nucleobases, nucleobase protected by protecting groups capable of withstanding the deprotection conditions of 5 'position of a nucleotide is preferred.
[0076]
　As the amino-protecting group is not particularly limited, for example, Greens Protective Groups in Organic Synthesis (Greene's PROTECTIVE GROUPS in ORGANIC SYNTHESIS), 4th edition, Wiley-Interscience (Wiley-Interscience ) publication (which can include a protective group that has been described in 2006), and the like. Specific examples of the protecting group, a pivaloyl group, pivaloyl acetoxyphenyl methyl group, acetyl group, trifluoroacetyl group, phenoxyacetyl group, 4-isopropyl phenoxyacetyl, 4-tert-butyl-phenoxyacetyl, benzoyl, isobutyryl group, and a (2-hexyl) decanoyl group, dimethylformamidine isoxazolidinyl group, 1- (dimethylamino) ethylidene group, 9-fluorenylmethyloxycarbonyl group. Among these, an acetyl group, phenoxyacetyl group, 4-isopropyl phenoxyacetyl group, benzoyl group, an isobutyryl group, a (2-hexyl) decanoyl group, dimethylformamidine isoxazolidinyl group, and 1- (dimethylamino) ethylidene group.
[0077]
　Carbonyl group of the nucleobase may also be protected. For example, phenol, 2,5-dichlorophenol, 3-chlorophenol, 3,5-dichlorophenol, 2-formylphenol, 2-naphthol, 4-methoxyphenol, 4-chlorophenol, 2-nitrophenol, 4-nitro phenol, 4-acetylamino-phenol, pentafluorophenol, 4-pivaloyl acetoxyphenyl benzyl alcohol, 4-nitro phenethyl alcohol, 2- (methylsulfonyl) ethanol, 2- (phenylsulfonyl) ethanol, 2-cyano ethanol, 2- ( trimethylsilyl) ethanol, dimethylcarbamoyl chloride, diethyl carbamic acid chloride, ethyl phenyl carbamic acid chloride, 1-pyrrolidine carbonyl chloride, 4-morpholine carboxylic acid chloride, diphenyl By reacting Rukarubamin acid chloride or the like, can be protected carbonyl group of the nucleobase. Here, the protecting group of the carbonyl group, it may not be necessary particularly introduced.
[0078]
　Nucleobase, in addition to the groups mentioned above, nucleobase optional substituents (e.g., halogen atom, an alkyl group, an aralkyl group, an alkoxy group, an acyl group, an alkoxyalkyl group, a hydroxyl group, an amino group, monoalkylamino, dialkylamino, carboxy, cyano, modified nucleic acid bases which is substituted 1-3 at an arbitrary position by nitro, etc.) (e.g., 8-Buromoadeniru group, 8-bromo-guanyl group, 5-Buromoshitoshiru group, 5- Yodoshitoshiru group , 5-bromouracil group, 5-iodo uracil group, 5-fluorouracil group, 5-Mechirushitoshiru group, 8-oxo-guanyl group, Hipokisanchiniru group) are also encompassed.
[0079]
　In the present specification, the "halogen atom", fluorine atom, chlorine atom, bromine atom or iodine atom.
[0080]
　As used herein, "alkyl (group)" may be either linear or branched. As the "alkyl (group)", include alkyl group having 1 or more carbon atoms, in particular when there is no limitation of the scope carbon atoms, preferably C 1-10 alkyl groups, more preferably C 1-6 alkyl groups, more preferably C 1-5 alkyl group. Preferable specific examples of the case where there is no limitation in the carbon number range, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec- butyl, tert- butyl, pentyl, hexyl, and the like, especially methyl, ethyl is preferred .
　As used herein, "C a-b is a" means the number of carbon atoms is a more b less (a, b is an integer).
[0081]
　In the present specification, the "aralkyl (group)" is, C 7-20 an aralkyl group, preferably a C 7-16 aralkyl group (C 6-10 aryl -C 1-6 alkyl group). Suitable examples include benzyl, 1-phenylethyl, 2-phenylethyl, 1-phenylpropyl, naphthylmethyl, 1-naphthylethyl, 1-naphthyl-propyl, and the like, especially benzyl is preferred.
[0082]
　As used herein, "alkoxy (group)" may be either linear or branched. The "alkoxy (group)", include one or more alkoxy groups having a carbon number, especially when there is no limitation of the scope carbon atoms, preferably C 1-10 alkoxy groups, more preferably C 1-6 an alkoxy group. Preferable specific examples of the case where there is no limitation in the carbon number range, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec- butoxy, tert- butoxy, pentyloxy, hexyloxy and the like, in particular methoxy, ethoxy is preferred.
[0083]
　As used herein, "acyl (group)" may be either linear or branched. As the "acyl (group)", for example, C 1-6 alkanoyl group, C 7-13 aroyl group, and the like. Specifically, for example, formyl, acetyl, n- propionyl, isopropionyl, n- butyryl, isobutyryl, pivaloyl, valeryl, hexanoyl, benzoyl, naphthoyl, levulinyl like can be mentioned, which may be substituted, respectively.
[0084]
　As used herein, "alkenyl (group)" may be either linear or branched. "Alkenyl (group)", for example, C 2-6 and an alkenyl group. Specifically, for example, vinyl, 1-propenyl, allyl, isopropenyl, butenyl, isobutenyl, and the like. Among them, C 2-4 alkenyl group are preferred.
[0085]
　As used herein, "alkynyl (group)" may be either linear or branched. "Alkynyl (group)", for example, C 2-6 an alkynyl group. Specifically, for example, 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. Among them, C 2-4 alkynyl group.
[0086]
　As used herein, "cycloalkyl (group)" means a cyclic alkyl group, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. Of these, cyclopropyl, cyclobutyl, cyclopentyl, C such as cyclohexyl 3-6 preferably a cycloalkyl group, cyclohexyl is particularly preferred.
[0087]
　As used herein, "aryl (group)" means a hydrocarbon radical of a monocyclic or polycyclic showing aromaticity (condensation), specifically, for example, phenyl, 1-naphthyl, 2 - naphthyl, biphenylyl, C and 2-anthryl 6-14 aryl group and the like. Among them C 6-10 , more preferably an aryl group, and particularly preferably phenyl.
[0088]
　In the present specification, the "hydrocarbon group", for example, aliphatic hydrocarbon groups, aromatic aliphatic hydrocarbon group, monocyclic saturated hydrocarbon group and an aromatic hydrocarbon group and the like, specifically , for example, an alkyl group, an alkenyl group, an alkynyl group, a cycloalkyl group, an aryl group, monovalent and divalent radicals derived therefrom, such as aralkyl groups.
[0089]
　As used herein, "C 6-14 hydrocarbon ring", for example, C 6-10 cycloalkane, C 6-10 cycloalkene, C 6-14 an aromatic hydrocarbon ring.
　The "C 6-10 The cycloalkane", for example, cyclohexane, cycloheptane, and cyclooctane.
　The "C 6-10 The cycloalkene", for example, cyclohexene, cycloheptene, and cyclooctene.
　The "C 6-14 aromatic hydrocarbon ring", for example, benzene, naphthalene.
[0090]
　As used herein, "alkylene (group)" may be either linear or branched. As the "alkylene (group)", include an alkylene group having 1 or more carbon atoms, in particular when there is no limitation of the scope carbon atoms, preferably C 1-10 alkylene group, more preferably C 1-6 is an alkylene group. Preferable specific examples include methylene, ethylene, propylene, butylene, pentylene, hexylene and the like, especially methylene, ethylene is preferred.
　In the present specification, the "linker", for example, -O -, - C (= O) -, - C (= O) O -, - OC (= O) -, - C (= O) NH- , -NHC (= O) -, - S -, - SO -, - SO 2 -, - Si (R ') (R ") O -, - Si (R') (R") - (R ', R "are each independently hydrogen atom or C 1-22 a hydrocarbon group.), and the like.
[0091]
　Herein, the "substituent" in "optionally substituted", the halogen atom, alkyl group, aralkyl group, an alkoxy group, an acyl group, an alkenyl group, an alkynyl group, a cycloalkyl group, an aryl group other, hydroxyl, (as with the alkyl portion wherein the alkyl group) nitro, (same alkoxyl portion of the alkoxy group) a cyano group, guanidyl group, a carboxy group, an alkoxycarbonyl group, a sulfo group, phospho group, an alkylthio group, alkylsulfinyl group (like alkyl portions of the alkyl group), an alkylsulfonyl group (the alkyl part as well as the alkyl group), an amino group, monoalkylamino group (the alkyl part as well as the alkyl group), a dialkylamino group ( alkyl portion as well as the alkyl group), such as oxo group and the like.
[0092]
[Oligonucleotide production method]
　the production method of the present invention, (sometimes abbreviated hereinafter as "3'-5 'synthesis") mode that performs direction of oligonucleotide chain elongation to the 5' end of the 3 'end and it includes both 5 embodiment in the direction of 'the end 3' to the end performs extension of the oligonucleotide chain (hereinafter occasionally abbreviated as "5'-3 'synthesis"). It will be described first manufacturing method of the present invention which is a 3'-5 'synthesis.
[0093]
3'-5 'synthesis
　production method of the present invention which is a 3'-5' synthesis, the following steps (1), including (3), (4) and (6). The manufacturing method optionally further following step (2) may contain (5) and (7).
　(1) in a non-polar solvent,
　5 'hydroxyl group is not protected, an amino group and imino group nucleobases, 2 of ribose residue' position hydroxyl, 3'-position hydroxyl group and 3'-amino group, as well as deoxy at least one group selected from 3 'position hydroxyl and 3'-amino group of the ribose residue is not removed under acidic conditions, it is protected with a removable protecting group under basic conditions, are and other groups better nucleosides be protected by protecting groups used in nucleic acid synthesis, or oligonucleotide (a), or
　5 'position hydroxyl group is not protected, the 3'-position a hydroxyl group of phosphate group -OL n1 (wherein, L -OH n1 represents an organic group.) is replaced by, -OL n1 hydroxyl group of -OH is not removed under acidic conditions, are protected with a removable protecting group under basic conditions ,and Other groups are further protected may be substituted or oligonucleotide with a protecting group to be used in nucleic acid synthesis (alpha)
to the reaction solution containing,
　3 'hydroxyl group or 3' amino group is phosphoramidite of 5'-position hydroxyl group is protected by a temporary protecting group which can be removed under acidic conditions, and other groups are further not removed under acidic conditions may nucleosides be protected by a protecting group selected from the protecting groups used in the removable protecting group and nucleic acid synthesis under basic conditions, or oligonucleotide (b)
was added to a nucleoside, nucleotide or oligonucleotide nucleotides (a) or a substituted or oligonucleotide (alpha) and a nucleoside, nucleotide or oligonucleotide (b) and condensing, phosphite tri 5 'position hydroxyl group is protected with a temporary protecting group which can be removed under acidic conditions obtaining a reaction liquid containing ester of (c);
　(2) if necessary, query the reaction mixture after the condensation Step adding inches agent;
　(3) adding an oxidizing agent or a sulfurizing agent to a reaction solution containing phosphite triester body (c), by oxidation or sulfurization phosphite triester body (c), 5 'position hydroxyl obtaining a reaction solution containing the protected oligonucleotide with a temporary protecting group which can be removed under acidic conditions (d);
　was added to (4) acid to the reaction mixture after oxidation or sulfurization 5 'position of the hydroxyl group removing temporary protective group, the 5'-position step hydroxyl obtain a reaction solution containing an oligonucleotide (e) unprotected;
　(5) if necessary, adding a base to the reaction solution containing oligonucleotides (e) step to to neutralize;
　step (6) adding a polar solvent to a reaction solution containing oligonucleotides (e), purified by solid-liquid separation or extraction of the oligonucleotides (e); and
　(7) If necessary, resulting after removing all the protecting groups of an oligonucleotide, isolating the oligonucleotide unprotected.
[0094]
　Step (1), (3), (4) by repeating and the cycle of (6) (preferably the step (1) to (6)), it is possible to extend the oligonucleotide chain. The step (1), (3), (4) and a manufacturing method of an oligonucleotide comprising (6) are encompassed by the present invention.
[0095]
Step (1) (condensation)
　In this step, the 5'-position hydroxyl group is not protected, 3 'at least one position hydroxyl group is not removed under acidic conditions, removable protective groups under basic conditions in protected nucleoside, a nucleotide or an oligonucleotide (a) or a substituted or oligonucleotide (alpha), 3 'hydroxyl group or 3' amino group is phosphoramidite of 5'-position hydroxyl group under acidic conditions protected with a removable temporary protective group, and other groups are further protected with a protecting group under acidic conditions not removed, it is selected from protecting groups to be used in removable protecting group and nucleic acid synthesis under basic conditions good nucleosides be, by condensing a nucleotide or oligonucleotide (b), 5 'position hydroxyl group is protected with a temporary protecting group which can be removed under acidic conditions A step of obtaining scan triester body (c).
[0096]
　In a preferred embodiment of the process,
　the 5'-position hydroxyl group is not protected, an amino group and imino group nucleobases, 2 of ribose residue 'position hydroxyl group and 3'-position a hydroxyl group, and 3 of the deoxyribose residues' position at least one group selected from hydroxyl group, not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and other groups have been further protected by protecting groups used in nucleic acid synthesis which may nucleoside, nucleotide or oligonucleotide (a), or
　5 'position hydroxyl group is not protected, the 3'-position a hydroxyl group -OL phosphate group n1 of -OH (wherein, L n1 of the organic group shown. are replaced), -OL n1 hydroxyl group of -OH is not removed under acidic conditions, it is protected with a removable protecting group under basic conditions, and other groups are further nucleic acid synthesis It need be protected group protected or unprotected substituent or oligonucleotide (alpha), and
　3'-position a hydroxyl group is phosphoramidite of 5'-position hydroxyl group is protected with a temporary protecting group which can be removed under acidic conditions and other groups are further not removed under acidic conditions, may nucleosides be protected by a protecting group selected from the protecting groups used in the removable protecting group and nucleic acid synthesis under basic conditions, nucleotides or oligonucleotides (b)
it is used.
[0097]
　L n1 The organic group, a hydrocarbon group or a carbon atom in the hydrocarbon group, is meant a group replaced with a heteroatom. The hetero atom such as oxygen atom, nitrogen atom, sulfur atom and the like. Further, the organic group, a hydroxyl group, an amino group, which may have a substituent such as oxo group (= O). Hydroxyl and amino group organic group may have is preferably protected by a protecting group. The shape of the organic group is a chain (straight or branched), it may be either cyclic or a combination thereof.
[0098]
　The organic group may have a group having a function to the cell. Groups having a functional against cells, it is preferably bound to a terminal of the main chain or side chain of the organic group. Examples of the group having the function to the cell, for example, "By improving the lipid-soluble compound, based on improving the cell membrane permeability of the compound", "uptake through cell membrane receptor compounds into cells group "and the like to improve. "By improving the lipid-soluble compounds, cell membrane permeability to a group for improving the compound" includes, for example, cholesterol residue, and tocopherol residues and the like. As the "group to improve the uptake through cell membrane receptor compounds into cells", for example, N- acetylgalactosamine residues, and the like.
[0099]
　-OL n1 Examples of -OH,
[0100]
[Chemical Formula 17]

[0101]
　L n1 is preferably C 2-6 alkylene group, more preferably an ethylene group.
[0102]
　In a more preferred embodiment of this process,
　5'-position hydroxyl group is not protected, an amino group and imino group nucleobases, 2 of ribose residue 'position hydroxyl group and 3'-position a hydroxyl group, and 3 of the deoxyribose residues' at least one of the groups-position selected from hydroxyl, not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and other groups have been further protected by protecting groups used in nucleic acid synthesis which may be a nucleoside or oligonucleotide (a), and
　3 'position hydroxyl group is phosphoramidite of 5'-position hydroxyl group is protected with a removable temporary protective group under acidic conditions, and other groups more nucleic acid synthesis optionally protected nucleoside or oligonucleotide (b) with a protecting group to be used in
is used.
[0103]
　Concentration in the solution of the nucleoside to be used in this step, or oligonucleotide (a) or a substituted or oligonucleotide (alpha) is they are not particularly limited as long as dissolved in a solvent, preferably from 1 to 30 weight it is%.
[0104]
　Nucleoside, an amino group of a nucleotide or oligonucleotide (a), substituted or oligonucleotide (alpha) and a nucleoside, nucleotide or oligonucleotide (b) is preferably protected by a protective group described above. As the protective group, cetyl group, phenoxyacetyl group, 4-isopropyl phenoxyacetyl group, benzoyl group, an isobutyryl group, (2-hexyl) decanoyl group, dimethylformamidine isoxazolidinyl group, and 1- (dimethylamino) ethylidene group preferable. Nucleoside, if or oligonucleotide (a) and nucleoside, nucleotide or oligonucleotide (b) has a plurality of amino groups, the amino protecting group may be used alone or as two or more kinds.
[0105]
　Nucleoside, nucleotide or oligonucleotide (a) an amino group and imino group nucleobases, 2'-position hydroxyl group, 3 'of the ribose residue position hydroxyl group and 3'-amino group, and 3 of the deoxyribose residues' position hydroxyl group and 3 'position at least one group selected from amino groups, not removed under acidic conditions, are protected with a removable protecting group under basic conditions. Nucleoside, nucleotide or oligonucleotide (a) an amino group and imino group nucleobases, 3'-position hydroxy group, and 3 of the deoxyribose residues' of ribose residue at least one group selected from the position hydroxyl group, the protecting preferably protected by groups, position hydroxyl 3 'position hydroxyl group or deoxyribose residues 3' ribose residue is more preferably protected by the protective group.
[0106]
　Substituted or oligonucleotide (alpha) is, -OL n1 hydroxyl group of -OH is not removed under acidic conditions, are protected with a removable protecting group under basic conditions.
[0107]
　Nucleoside, nucleotide or oligonucleotide (a) and substituted or oligonucleotide (alpha) has "not removed under acidic conditions, the protecting group can be removed under basic conditions", as well as nucleosides, nucleotides or oligonucleotides (b ) is "not removed under acidic conditions, removable protective groups under basic conditions" which may have each independently the step (6) (solid-liquid separation or extraction) efficiently to do have at least one aliphatic hydrocarbon group having or having a linear aliphatic hydrocarbon group having 10 or more carbon atoms, or branched chain 1 or more, and the total carbon number of 14 to 300 it is preferable that a protecting group having an organic group is (hereinafter sometimes abbreviated as "anchor").
[0108]
　Anchor, nucleoside, nucleotide or oligonucleotide (a) and substituted or oligonucleotide (alpha) (optionally a nucleoside, nucleotide or oligonucleotide (b)) to impart hydrophobicity to, improve solubility to non-polar solvent make. It is also possible to reduce the solubility in polar solvents. Such protected nucleoside with an anchor, or oligonucleotide (a) and substituted or oligonucleotide (alpha) may be carried out condensation reaction in the liquid phase of a non-polar solvent, followed by step (6) in by adding a polar solvent to the reaction solution, protected oligonucleotide anchor (e) precipitated, it is possible to perform the solid-liquid separation. Or step (6) by adding a polar solvent to the reaction mixture in a polar solvent - is separated into layers between non-polar solvent, by shifting the oligonucleotides (e) in a non-polar solvent, it is possible to perform the extraction . Such anchors, for example, may be used those described in Patent Document 1, Patent Document 2 and WO 2013/122236.
[0109]
　If Purification by solid-liquid separation in step (6), the anchor is preferably a protecting group having a linear aliphatic hydrocarbon group having 10 or more carbon, purification by extraction in step (6) when performing the anchor has at least one aliphatic hydrocarbon group having a branched chain 1 or more, and it is preferable that the total carbon number of protecting groups having an organic group is 14 or more 300 or less.
[0110]
　First, a description will be given of a preferred anchor for solid-liquid separation. C Suitable anchor for solid-liquid separation, for example, a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker attached through a linker 6 -14 include protecting groups having a hydrocarbon ring.
[0111]
　The aliphatic hydrocarbon group having a carbon number of 10 or more linear shape, preferably linear C 10-40 alkyl group and a linear C 10-40 is a group selected from an alkenyl group, more preferably linear C 10-40 alkyl group, more preferably straight-chain C 10-30 alkyl group, particularly preferably a linear C 12-28 alkyl group, and most preferably a straight-chain Jo of C 14-26 alkyl group.
[0112]
　The linker is preferably -O -, - C (= O ) -, - C (= O) O -, - OC (= O) -, - C (= O) NH -, - NHC (= O) -, - S -, - SO -, - SO 2 -, and -Si (R ') (R " ) O -, - Si (R') (R") - (R ', R " are each independently and, a hydrogen atom or a C 1-22 selected from a hydrocarbon group), more preferably -O -., - C (= O) -, - C (= O) O -, - OC (= O ) -, - C (= O ) NH- and -NHC (= O) - is selected from, more preferably -O-.
[0113]
　The C 6-14 hydrocarbon ring is preferably selected from a benzene ring, a naphthalene ring and a cyclohexane ring, and more preferably selected from a benzene ring and cyclohexane ring, more preferably a benzene ring.
[0114]
　Suitable anchor for solid-liquid separation is preferably straight-chain C 10-40 benzene ring hydrocarbon group in which the alkyl group is linked via a single bond or -O- is bonded via -O- it is a protecting group with.
[0115]
　Next, preferred anchor will be described for the extraction. "Having at least one aliphatic hydrocarbon group having 1 or more branched-chain, and an organic group has a total carbon number of 14 to 300." The "branched" in a linear or branched chain saturated an aliphatic hydrocarbon group, C 1-6 preferably an alkyl group, C 1-4 more preferably an alkyl group, more preferably a methyl group or an ethyl group. Further, the "branched chain" may be substituted with one or more halogen atoms.
[0116]
　"Having at least one aliphatic hydrocarbon group having 1 or more branched-chain, and an organic group has a total carbon number of 14 to 300." The "aliphatic hydrocarbon group" in a linear, saturated or an aliphatic unsaturated hydrocarbon group, C 2-300 alkyl groups (preferably, C 3-100 alkyl groups, more preferably, C 3-60 alkyl group), C 2-300 alkenyl group (preferably, C 3-100 alkenyl group, more preferably, C 3-60 alkenyl group) or C 2-300 alkynyl group (preferably, C 3-100 alkynyl groups, more preferably, C 3-60 alkynyl group).
[0117]
　Site "has at least one aliphatic hydrocarbon group having branched chain 1 or more, and an organic group has a total carbon number of 14 to 300." "aliphatic hydrocarbon group having a branched chain one or more" in the is not particularly limited, (monovalent groups) be present at the terminal, which may (e.g. divalent) present in the other portion.
[0118]
　As the "aliphatic hydrocarbon group having 1 or more branched", for example, propyl group, butyl group, pentyl group, hexyl group, heptyl group, octyl group, nonyl group, decyl group, undecyl group, dodecyl group (lauryl group ), tridecyl group, myristyl group, cetyl group, stearyl group, arachidyl group, behenyl group, oleyl group, linolyl group, a branched isomer of such lignoceryl group, a monovalent and their having one or more branched divalent groups derived thereof. "Aliphatic hydrocarbon group having 1 or more branched-chain" preferably, 3,7,11-trimethyl-dodecyl group, 3,7,11,15-tetramethyl-hexadecyl group (hereinafter, 2,3 Jihidorofi sometimes referred butyl group.), a 2,2,4,8,10,10- hexamethyl undecane-5-yl group.
[0119]
　"Having at least one aliphatic hydrocarbon group having 1 or more branched-chain, and an organic group has a total carbon number of 14 to 300." "aliphatic hydrocarbon group having branched chain 1 or more" in several when present, each be the same, may be different.
[0120]
　"Having at least one branched chain aliphatic hydrocarbon group having 1 or more, and a total carbon number organic group is 14 or more 300 or less" in the non "branched chain aliphatic hydrocarbon group having 1 or more" sites can be arbitrarily set. For example -O -, - S -, - CO -, - NH -, - COO -, - OCONH -, - CONH -, - NHCO-, hydrocarbon group (monovalent or divalent) have a site, such as it may be in. As the "hydrocarbon group", for example, aliphatic hydrocarbon groups, aromatic aliphatic hydrocarbon group, monocyclic saturated hydrocarbon group and an aromatic hydrocarbon group and the like, specifically, for example, an alkyl group , an alkenyl group, an alkynyl group, a cycloalkyl group, an aryl group, monovalent groups and divalent groups derived from these, such as an aralkyl group is used. As the "alkyl group", C 1-6 preferably an alkyl group, e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec- butyl, tert- butyl, pentyl, hexyl and the like. As the "alkenyl group", C 2-6 preferably an alkenyl group such as vinyl, 1-propenyl, allyl, isopropenyl, butenyl, isobutenyl, and the like. As the "alkynyl group", C 2-6 alkynyl group are preferred, for example, ethynyl, propargyl, 1-propynyl, and the like. As the "cycloalkyl group", C 3-6 cycloalkyl group is preferred, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl. "Aryl group", C 6-14Aryl group are preferred, for example, phenyl, 1-naphthyl, 2-naphthyl, biphenylyl, 2-anthryl and the like. Among them C 6-10 , more preferably an aryl group, and particularly preferably phenyl. The "aralkyl group", C 7-20 preferably an aralkyl group, for example, include benzyl, 1-phenylethyl, 2-phenylethyl, 1-phenylpropyl, naphthylmethyl, 1-naphthylethyl, 1-naphthyl-propyl and the like It is. Among them, C 7-16 aralkyl group (C 6-10 aryl -C 1-6 alkyl group) is more preferable, benzyl is especially preferred. The "hydrocarbon group", halogen atom (chlorine atom, bromine atom, fluorine atom, iodine atom), may be substituted with a substituent selected from oxo group and the like.
[0121]
　"Having at least one branched chain aliphatic hydrocarbon group having 1 or more, the organic group and the total carbon number of 14 to 300." "total number of carbon atoms" in the 14 or more, preferably 16 or more, more preferably at least 18, 300 or less, preferably 200 or less, more preferably 160 or less. Further, "having at least one aliphatic hydrocarbon group having a branched chain 1 or more, and an organic group has a total carbon number of 14 to 300." The number of branch chains in is not particularly limited, preferably 2 or more , more preferably 3 or more, more preferably 4 or more, more preferably 8 or more, more preferably 10 or more. The larger the number of the branched chain is large, even if the oligonucleotide chain becomes long chain, having at least one aliphatic hydrocarbon group having a branched chain 1 or more, and the total number of carbon atoms is 14 to 300 nucleoside or oligonucleotide protected with an anchor having an organic group, the organic solvent (in particular, non-polar solvent) is readily soluble respect.
[0122]
　The "branched chain having at least one aliphatic hydrocarbon group having 1 or more, and an organic group has a total carbon number of 14 to 300", the formula (A):
[0123]
[Chem. 18]

[0124]
Wherein
　* indicates the bonding position to adjacent
　atoms; R 14 and R 15 are each independently hydrogen atom or C 1-4 an alkyl group; and
　X 1 is a single bond or C 1 -4 an alkylene group.
　However, R 14 and R 15 are not both are hydrogen atoms. ]
Groups have the same or different divalent group represented by are preferred.
[0125]
　Examples of the group having a divalent group represented by the formula (A), for example, groups represented by any one of the following formulas (B) ~ (D). Incidentally, formula (B) ~ carbon atoms in the definition of each symbol in the (D), the number of repeating units (m 1 , n 0 ~ n 2 be one) like that shown for convenience, the total carbon number of 14 or more (preferably 16 or more, more preferably 18 or higher), 300 or less (preferably 200 or less, more preferably 160 or less) can be appropriately modified within the scope of the definitions described above so as to be. Hereinafter, formula (B) ~ (D), will be described in order.
[0126]
　Formula (B) is as follows.
[0127]
[Of 19]

[0128]
Wherein
　* indicates the bonding position to adjacent
　atoms; R 16 and R 17 are either a hydrogen atom, or together = indicates
　O; n 0 is an integer of 2 to 40 ;
　n 0 one R 18 and R 19 are each independently hydrogen atom or C 1-4 an alkyl
　group; n 0 one X 2 are each independently a single bond or C 1-4 alkylene It represents a
　group; R 20 represents a hydrogen atom or a C 1-4 an alkyl group; and
　R 21 is C 1-4 an alkyl group.
　However, R 18And R 19 never are both hydrogen atom, and n 0 if is 2, R 20 is C 1-4 an alkyl group. ]
[0129]
　The group of formula
　(B), R 16 and R 17 is a are both hydrogen
　atom; n 0 is an 2-40
　integer; n 0 one R 18 and R 19 are each, independently, hydrogen atom, a methyl group or an ethyl group;
　n 0 one X 2 are each independently a single bond, is methylene group or an ethylene group; and
　R 20 is a hydrogen atom, is a methyl group or an ethyl group
group (wherein, R 18 and R 19 never are both hydrogen atom, and n 0 if is 2, R 20 represents a methyl or ethyl group.) is preferred.
[0130]
　More group preferred formula (B) is a myristyl group, cetyl group, stearyl group, an arachidyl group, branched isomers of carbon number of 14 to 160 such as behenyl groups, among them 2,3-dihydro-phytyl group, 3 , 7,11-trimethyl-dodecyl group, 2,2,4,8,10,10- hexamethyl-5-dodecanoyl group is particularly preferred.
[0131]
　Formula (C) are as follows.
[0132]
[Of 20]

[0133]
Wherein
　* indicates the bonding position to adjacent atoms;
　m 1 pieces of OR 22 each independently represents a hydroxyl group which is substituted by a group represented by the formula (B); and
　m 1 is represents an integer of 1-3. ]
　Incidentally, the expression "group of the formula (B)" in (C), the * is O (i.e., adjacent atoms) except that indicate a point of attachment to be as described above.
[0134]
　In the group of formula (C), R 22 is a myristyl group, cetyl group, stearyl group, arachidyl group, more preferably branched isomers group having 14 to 30 carbon atoms such as behenyl groups, among them 2,3-dihydro phytyl group, 3,7,11-trimethyl-dodecyl group is particularly preferable.
[0135]
　Formula (D) are as follows.
[0136]
[Of 21]

[0137]
Wherein
　* indicates the bonding position to
　Q; n 1 represents an integer of 1 ~
　10; n 2 represents an integer of 1 ~
　10; n 1 one R 26 and R 27 are, each independently represents a hydrogen atom or a C 1-4 an alkyl
　group; n 1 single X 3 are each independently a single bond or a C 1-4 an alkylene
　group; n 2 pieces of R 28 and R 29 are each independently hydrogen atom or C 1-4 an alkyl
　group; n 2 pieces of X 5 are each independently a single bond or C 1-4 represents an alkylene
　group; X 4 is a single bond or a C 1-4 an alkylene group; and
　R 23 , R 24 , R 25 , R 30 , R 31 and R 32 are each independently hydrogen atom or C 1-4 an alkyl group.
　However, R 26 and R 27 , and / or R 28 and R 29 never are both hydrogen atom, and n 1 + n 2 if is 2, R 23 , R 24 and R 25 2 or more but each independently, C 1-4 represents an alkyl group, or R 30 , R 31 and R 32 2 or more are each independently of, C 1-4 an alkyl group. ]
[0138]
　The group of formula
　(D), n 1 is an There integer of 1 ~
　5; n 2 is an integer of 1 ~
　5; n 1 one R 26 and R 27 are each independently hydrogen atom, a methyl group or an ethyl
　group; n 1 single X 3 are each independently a single bond, a methylene group or an ethylene
　group; n 2 pieces of R 28 and R 29 are each independently, hydrogen atom, with there methyl group or an ethyl group;
　n 2 pieces of X 5 are each independently a single bond, a methylene group or an ethylene group;
　X 4 is a single bond, a methylene group or an ethylene group; and
　R 23, R 24 , R 25 , R 30 , R 31 and R 32 are each independently a hydrogen atom or a C 1-4 alkyl group
group (wherein, R 26 and R 27 , and / or R 28 and R 29 is that they are not both hydrogen atoms, and n 1 + n 2 if is 2, R 23 , R 24 and R 25 are two or more of each independently, C 1-4 represents an alkyl group or R, 30 , R 31 and R 32 2 or more are each independently, C 1-4It represents an alkyl group. ) Is more preferable.
[0139]
　Particularly group of suitable Formula
　(D), n 1 is 1 to be a 5
　integer; n 2 is an integer of 1 to
　5; n 1 one R 26 and R 27 are each independently Te is a hydrogen atom or a methyl
　group; n 1 single X 3 are each independently a single bond or a methylene
　group; n 2 pieces of R 28 and R 29 are each independently hydrogen atom or a methyl
　group; n 2 pieces of X 5 are each independently a single bond or a methylene
　group; X 4 is a single bond or a methylene group; and
　R 23 , R 24 , R 25 , R 31 , R 31 and R 32 are methyl
group (wherein, R 26 and R 27 and / or R, 28 and R 29 are never both hydrogen atoms) include It is.
[0140]
　"Having at least one aliphatic hydrocarbon group having 1 or more branched-chain, and the total carbon number of the organic group is 14 or more 300 or less" Specific examples of include the following groups. * In each group indicates the bonding position; n in Formula 3 represents an integer of 3 or more; n 4 , the total number of carbon atoms of the group may be appropriately set to be 14 to 300.
[0141]
[Formula 22]

[0142]
　"Having at least one branched chain aliphatic hydrocarbon group having 1 or more, and a total carbon number organic group is 14 or more 300 or less" Preferred examples of include the following groups:
　3,7, 11,15- tetramethyl-hexadecyl group (also known as 2,3-dihydro-phytyl
　group); 3,7,11-trimethyl-dodecyl
　group; 2,2,4,8,10,10- hexamethyl-5-dodecanoyl ;
　3,4,5 (3 ', 7', 11 ', 15'- tetramethyl-hexadecyl) benzyl group; and
　3,5-di (3', 7 ', 11', 15'- tetra methyl hexadecyl) benzyl group.
[0143]
　Anchor, more preferably (sometimes hereinafter abbreviated as "anchor (g-I)".) A group represented by the following formula (g-I).
L-Y **-Z (g-I)
[wherein,
　** represents the bonding position of the base to be protected;
　L is a single bond, or the formula (a1) or (a1 '):
[0144]
[Of 23]

[0145]
(Wherein,
　* indicates the bonding position to
　Y; ** is an defined as
　above; R 1 and R 2 are each independently, C 1-22 a hydrocarbon
　group; L 1 is optionally substituted divalent C 1-22 represents a hydrocarbon
　group; L 2 is either a single bond, or C *** (= O) N (R 3 ) -R 4 -N (R 5 ) **** (wherein, *** is L 1 represents a bonding position with, **** denotes the bonding position to C = O, R 4 is C 1-22 alkylene group are shown, R 3 and R 5 are each independently a hydrogen atom or a C 1-22 represents an alkyl group, or R 3 and R 5 are taken together, may form a ring. ) A group represented by. A group represented by);
　Y is a single bond, an oxygen atom or NR (R, represents a hydrogen atom, an alkyl group or an aralkyl group); the. And
　Z has the formula (a2), the formula ( a2 ') or formula (a2 "):
[0146]
[Of 24]

[0147]
Wherein
　* indicates the binding position;
　R 6 is a hydrogen atom, or R b is, when a group represented by the following formula (a3), the ring A or ring B R 6 is R 8 represents a single bond or -O- together with ring a or ring B and ring C may form a fused ring with;
　k is an integer of 1 to 4;
　the k Q are each independently, -O -, - C (= O) -, - C (= O) O -, - OC (= O) -, - C (= O) NH- or - NHC (= O) - are shown;
　k-number of R 7 each independently represent a hydrocarbon group is a linear aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker or branched chain having at least one aliphatic hydrocarbon group having 1 or more, and the total number of carbon atoms represents an organic group is 14 or more 300 or less;
　ring a Contact And ring B are each independently, k-number of QR 7 in addition to further halogen atom, a C substituted with a halogen atom 1-6 optionally substituted alkyl group, and a halogen atom C 1-6 may have a substituent group selected from the group consisting of an alkoxy group;
　R a represents a hydrogen atom; and
　R b is a hydrogen atom or the formula, (a3):
[0148]
[Of 25]

[0149]
(Wherein,
　* indicates the bonding position;
　j is 0 to an integer of 4;
　j-number of Q are each independently a as defined above;
　j-number of R 9 is each independently Te, at least one organic aliphatic hydrocarbon radical having either a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker, or branched one or more of and, and the total number of carbon atoms is an organic group of 14 to 300;
　R 8 is either a hydrogen atom, or R of ring a or ring B 6 taken together with a single bond or -O- Te, ring a or ring B and ring C may form a fused ring with; and
　ring C, j-number of QR 9 in addition to further halogen atom, a C substituted with a halogen atom 1 -6 alkyl groups, and may be substituted by a halogen atom C 1- May have a substituent group selected from the group consisting of an alkoxy group.) In either a group represented, or
　R a and R b are taken together to form an oxo group. It shows a group represented by. ]
[0150]
　Equation (a2), R in the formula (a2 ') and the formula (a2 ") 7 , and R in the formula (a3) 9 aliphatic hydrocarbon group having a carbon number of 10 or more straight chain having each independently to, preferably linear C 10-40 alkyl group and a linear C 10-40 is a group selected from an alkenyl group, more preferably linear C 10-40 alkyl group, further preferably straight-chain C 10-30 alkyl group, particularly preferably straight-chain C 12-28 alkyl group, most preferably straight-chain C 14-26 alkyl group.
　formula (a2 ), the formula (a2 ') and wherein R (a2 ") 7 , and R in the formula (a3) 9 linker has are each independently preferably -O -, - C (= O ) - , -C (= O) O - , - OC (= O) -, - C (= O) NH- or It is -NHC (= O) - and is, more preferably -O-.
[0151]
　Equation (a2), the formula (a2 ') and the formula (a2 ") R in 7 , and the formula (a3) in R 9 of the" aliphatic hydrocarbon group having 10 or more straight is a single bond or hydrocarbon group "bonded via a linker, preferably straight-chain C 10-40 alkyl group, one to three linear C 10-40 alkyl group is linked via a -O- benzyl group or 1-3 linear C, 10-40 cyclohexylmethyl group wherein the alkyl group is linked via a -O-.
[0152]
　Equation (a2), the formula (a2 ') and the formula (a2 ") in R 7 , and R in the formula (a3) 9 aliphatic hydrocarbon group of at least one having an embodiment of" branched one or more One has, and organic group "total carbon number of 14 to 300 are each independently preferably a group having a divalent group represented by the above formula (a), more preferably the a group represented by any one of formula (B) ~ (D), more preferably a group represented by the formula (B), particularly preferably 2,3-dihydro phytyl group, 3,7 a 11-trimethyl-dodecyl group or 2,2,4,8,10,10- hexamethyl-5-dodecanoyl.

The scope of the claims
[Requested item 1]
The following steps (1), (3), (4) and (6) a manufacturing method of an oligonucleotide comprising:
(1) the non-polar solvent,
　5 'hydroxyl group is not protected, the amino group of the nucleobase and imino group, the 2'-position hydroxyl group, 3 'of the ribose residue of at least one group selected position hydroxyl and 3'-amino group, and 3 of the deoxyribose residues' from position hydroxyl group and 3'-amino group, an acid not removed under the conditions, are protected with a removable protecting group under basic conditions, and optionally nucleosides be protected with a protecting group other groups are further used in nucleic acid synthesis, or oligonucleotide (a) or
　5 'position hydroxyl group is not protected, 3' one hydroxyl group in position phosphate group -OL n1 (wherein, L -OH n1 represents an organic group.) is replaced by, -OL n1 -OH of Group is not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and other groups are further protected may be substituted or oligonucleotide with a protecting group to be used in nucleic acid synthesis (alpha)
to the reaction solution containing,
　3 'hydroxyl group or 3' amino group is phosphoramidite of 5'-position hydroxyl group is protected with a temporary protecting group which can be removed under acidic conditions, is and other groups Furthermore, not removed under acidic conditions, basic conditions can be removed by a protecting group and may nucleosides be protected with a protecting group selected from the protecting groups used in nucleic acid synthesis, or oligonucleotide (b)
Was added and the nucleoside, nucleotide or oligonucleotide (a) or a substituted or oligonucleotide (alpha) and a nucleoside, nucleotide or oligonucleotide (b) and condensing, 5 'hydroxyl group which can be removed under acidic conditions obtaining a reaction solution containing the protected phosphite triester temporary protecting group (c);
　(3) by adding an oxidizing agent or a sulfurizing agent to a reaction solution containing phosphite triester body (c), phosphite oxidized or sulfurized triester body (c), the step 5 'hydroxyl group to obtain a reaction liquid containing a protected oligonucleotide with a temporary protecting group which can be removed under acidic conditions (d);
　(4) oxidizing or by adding an acid to the reaction solution after sulfurization 5 'position is removed a temporary protecting group for a hydroxyl group, 5' oligonucleotide-position hydroxyl group is not protected (E) obtaining a reaction solution containing; and
　(6) adding a polar solvent to a reaction solution containing oligonucleotides (e), purifying the solid-liquid separation or extraction of the oligonucleotides (e).
[Requested item 2]
　In step (1),
　5 'hydroxyl group is not protected, an amino group and imino group nucleobases, 2 of ribose residue' position hydroxyl group and 3'-position a hydroxyl group, and 3 of the deoxyribose residues' position hydroxyl group at least one group selected from and not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and other groups be further protected by protecting groups used in nucleic acid synthesis good nucleoside, nucleotide or oligonucleotide (a), or
　5 'position hydroxyl group is not protected, the 3'-position a hydroxyl group of phosphate group -OL n1 in -OH (wherein, L n1 represents an organic group . is replaced), -OL n1 hydroxyl group of -OH is not removed under acidic conditions, it is protected with a removable protecting group under basic conditions, and other groups are further used in nucleic acid synthesis coercive Protected substituted or nucleotides have or oligonucleotide group (alpha), and
　3'-position a hydroxyl group is phosphoramidite of 5'-position hydroxyl group is protected with a temporary protecting group which can be removed under acidic conditions, and other group is further not removed under acidic conditions, basic conditions can be removed by a protecting group and may nucleosides be protected with a protecting group selected from the protecting groups used in nucleic acid synthesis, or oligonucleotides ( b)
the method according to claim 1 that use.
[Requested item 3]
　In step (1),
　5 'hydroxyl group is not protected, an amino group and imino group nucleobases, 2 of ribose residue' position hydroxyl group and 3'-position a hydroxyl group, and 3 of the deoxyribose residues' position hydroxyl group at least one group selected from and not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and other groups be further protected by protecting groups used in nucleic acid synthesis good nucleoside or oligonucleotide (a), and
　3 'position hydroxyl group is phosphoramidite of 5'-position hydroxyl group is protected with a removable temporary protective group under acidic conditions, and using the other group is more nucleic acid synthesis optionally protected nucleoside or oligonucleotide (b) with a protecting group for
the production method according to claim 1 that use.
[Requested item 4]
　Prior to step (1) and the step (3) after the further steps of the manufacturing method according to any one of claims 1 to 3 comprising (2):
　(2) quench the reaction mixture after the condensation adding the agent.
[Requested item 5]
　Quenching agent used in step (2) is an alcohol The method of claim 4 is at least one selected from phenols and amines.
[Requested item 6]
　To the reaction solution before the step (1) and the step (4) after the mixture of carboxylic acids and organic bases, adding an inorganic acid or an amine, in the step (3), 5 as a sulfurizing agent - [(N, the process according to any one of claims 1 to 5, the addition of N- dimethylaminomethylidene) amino]-3H-1,2,4-dithiazole-3-thione in the reaction solution.
[Requested item 7]
　The amount of the basic nitrogen atoms of the organic base has the method according to claim 6 which is 1 to 2 mol with respect to the carboxy group 1 mol having a carboxylic acid.
[Requested item 8]
　Sulfurizing agent used in step (3) is, 3H-1,2-benzodithiol-3-one 1,1-dioxide, 3H-1,2-benzodithiole-3-one, phenylacetyl disulfide, tetraethyl thiuram disulfide , di-pentamethylene thiuram tetrasulfide, phenyl-3H-1,2,4-dithiazole-3-one, wherein at least one selected from 3-amino-1,2,4-dithiazole-5-thione and sulfur the process according to any one of claims 1-5.
[Requested item 9]
　Oxidizing agent used in step (3) is iodine, (1S) - (+) - (10- camphorsulfonic sulfonyl) oxaziridine, tert- butyl hydroperoxide, 2-butanone peroxide, 1,1-dihydroperoxy cyclododecane, bis (trimethylsilyl) peroxide and at least one manufacturing method according to any one of claims 1 to 5 is selected from m- chloroperbenzoic acid.
[Requested item 10]
　The process according to the temporary protecting groups, any one of claims 1 to 9, which is a dimethoxytrityl group or monomethoxytrityl group.
[Requested item 11]
　Nonpolar solvent, the production method according to halogenated solvents, any one of claims 1 to 10 is at least one selected aromatic solvent, an ester-based solvents and aliphatic solvents.
[Requested item 12]
　Acid used in step (4) is trifluoroacetic acid, dichloroacetic acid, trifluoromethanesulfonic acid, trichloroacetic acid, methanesulfonic acid, hydrochloric acid, at least is one claims 1 to selected from acetic acid and p- toluenesulfonic acid the process according to any one of 11.
[Requested item 13]
　Not removed under acidic conditions, a protective group removable under basic conditions, aliphatic hydrocarbon radical having either a straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms, or branched one or more at least one a, and process according to any one of claims 1 to 12 carbon atoms in total having an organic group is 14 or more 300 or less.
[Requested item 14]
　Step (6), and adding a polar solvent to a reaction solution containing oligonucleotides (e), according to any one of claims 1 to 13, which is a step of purifying the solid-liquid separation of oligonucleotides (e) Production method.
[Requested item 15]
　Polar solvent used in step (6) The production method according to any one of claims 1 to 14, which is a nitrile solvent.
[Requested item 16]
　Prior to step (4) and step (6) after the further steps of (5) The process according to any one of claims 1 to 15 including a:
　including (5) oligonucleotides (e) a step of neutralization by adding a base to the reaction solution.
[Requested item 17]
　Base used in step (5) is, pyridine, 2,4,6-trimethylpyridine, benzimidazole, 1,2,4-triazole, N- phenylimidazole, 2-amino-4,6-dimethyl pyrimidine, 1, 10- phenanthroline, imidazole, N- methylimidazole, 2-chloro-benzimidazole, 2-bromo-benzimidazole, 2-methylimidazole, 2-phenyl-benzimidazole, one at least selected from N- phenyl benzimidazole and 5-nitro-benzimidazole the method according to claim 16 One in which.
[Requested item 18]
　After step (6), further the following steps (7) The process according to any one of claims 1 to 17 comprising:
　(7) after removing any protecting group of the resulting oligonucleotides, protective isolating the oligonucleotide that are not.
[Requested item 19]
　Or the removal reaction of the temporary protecting group of step (4) carried out in the presence of a cation scavenger, or any one of claims 1 to 18 to add a cation scavenger to the reaction solution after removal reaction of the temporary protecting group the method according.
[Requested item 20]
　Cation scavenger, pyrrole, 2-methylpyrrole, 3-methylpyrrole, 2,3-dimethylpyrrole, 2,4-dimethylpyrrole, indole, 3-methylindole, 4-methylindole, 5-methylindole, 6 methylindole, 7-methylindole, 5,6-dimethyl indole, 6,7-dimethyl indole, 2-methylfuran, 2,3-dimethyl furan, 2-methyl-3- (methylthio) furan, and selected from menthofuran the method according to claim 19 is at least one.
[Requested item 21]
　The following steps (1 '), (3'), (4 ') and (6') oligonucleotides manufacturing method comprising:
　(1 ') in a non-polar solvent,
　3' position hydroxyl group or 3 'amino group not protected, an amino group and imino group nucleobases, 2'-position hydroxyl group and the 5'-position hydroxyl group of the ribose residue, and at least one group selected from the 5'-position hydroxyl group of deoxyribose residues, acidic conditions not removed under, it is protected with a removable protecting group under basic conditions, and other groups are better nucleosides be protected by protecting groups used in nucleic acid synthesis, or oligonucleotide (a ') or
　3 'position hydroxyl group or 3' amino group is not protected, one of the hydroxyl groups of 5 'position phosphate group -OL n1 -OH (wherein, L n1 replaced represents an organic group.) and, -OL N1 OH of the hydroxyl groups are not eliminated under acidic conditions, are protected with a removable protecting group under basic conditions, and other groups are further protected may be substituted nucleotides or oligonucleotides with a protecting group to be used in nucleic acid synthesis 'nucleotide (alpha)
to the reaction solution containing,
　5' hydroxyl group is phosphoramidite of, 3 'hydroxyl group or 3' amino group is protected with a temporary protecting group which can be removed under acidic conditions, and other group is further not removed under acidic conditions, basic conditions can be removed by a protecting group and may nucleosides be protected with a protecting group selected from the protecting groups used in nucleic acid synthesis, or oligonucleotides (b ')
Was added and the nucleoside, nucleotide or oligonucleotide (a ') or a substituted or oligonucleotide (alpha') and a nucleoside, nucleotide or oligonucleotide (b ') and the condensation of the 3' position hydroxyl group or 3 'amino ; 'groups obtaining a reaction liquid containing a phosphite triester protected with a removable temporary protective group under acidic conditions (c)
　a reaction solution containing (phosphite triester body (c') 3) ' by adding an oxidizing agent or a sulfurizing agent, phosphite triester (c ') the oxide or sulfide, 3' are protected at the a position hydroxyl group or 3 'amino group temporary protecting group which can be removed under acidic conditions to oligonucleotide 'to obtain a reaction liquid containing a step; (d)
　(4') 3 by adding an acid to the reaction mixture after oxidation or sulfurization 'position hydroxyl group or 3' temporary amino group The protecting group is removed, 3 'hydroxyl group or 3' amino groups oligonucleotides unprotected (e ') to obtain a reaction solution containing; and
　(6') reaction containing oligonucleotides (e ') step by adding a polar solvent, is purified by solid-liquid separation or extraction of the oligonucleotides (e ') to.
[Requested item 22]
　'In step (1)
　3' position hydroxyl group is not protected, an amino group and imino group nucleobases, 2'-position hydroxyl group and the 5'-position hydroxyl group of the ribose residue, as well as 5 'position of the deoxyribose residues at least one group selected from hydroxyl group, not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and other groups have been further protected by protecting groups used in nucleic acid synthesis which may nucleoside, nucleotide or oligonucleotide (a '), or
　3'-position hydroxy group is not protected, the 5'-position a hydroxyl group of phosphate group -OL n1 in -OH (wherein, L n1 of the organic group and replaced.) indicating, -OL n1 hydroxyl group of -OH is not removed under acidic conditions, it is protected with a removable protecting group under basic conditions, and other groups are further used in nucleic acid synthesis Protected substituted may be a nucleotide or oligonucleotide Mamorumoto (alpha '), as well as
　5' hydroxyl group is phosphoramidite of, 3'-position hydroxy group is protected with a temporary protecting group which can be removed under acidic conditions, and other groups are further not removed under acidic conditions, may nucleosides be protected by a protecting group selected from the protecting groups used in the removable protecting group and nucleic acid synthesis under basic conditions, nucleotide or oligonucleotide nucleotides (b ')
the method according to claim 21 used.
[Requested item 23]
　'In step (1)
　3' position hydroxyl group is not protected, the 5'-position a hydroxyl group of phosphate group -OL n1 (wherein, L -OH n1 is. Of an organic group) replaced by cage, -OL n1 hydroxyl group of -OH is not removed under acidic conditions, it is protected with a removable protecting group under basic conditions, and other groups have been further protected by protecting groups used in nucleic acid synthesis may also be substituted or oligonucleotide (alpha '), as well as
　5' hydroxyl group is phosphoramidite of, 3'-position hydroxy group is protected with a temporary protecting group which can be removed under acidic conditions, and other groups are further not removed under acidic conditions, basic conditions with a removable protecting group and optionally nucleosides be protected with a protecting group selected from the protecting groups used for nucleic acid synthesis or oligonucleotide (b ')
using a請 The process according to claim 21.
[Requested item 24]
　'In step (1)
　3' position hydroxyl group is not protected, an amino group and imino group nucleobases, 2'-position hydroxyl group and the 5'-position hydroxyl group of the ribose residue, as well as 5 'position of the deoxyribose residues at least one group selected from hydroxyl group, not removed under acidic conditions, are protected with a removable protecting group under basic conditions, and other groups have been further protected by protecting groups used in nucleic acid synthesis which may nucleoside or oligonucleotide (a '), as well as
　the 5' position hydroxyl phosphoramidite of, 3'-position hydroxy group is protected with a removable temporary protective group under acidic conditions, and other groups more nucleic acid synthesis optionally protected nucleoside or oligonucleotide (b ') with a protecting group used in the
manufacturing method according to claim 21 used.
[Requested item 25]
　Before the step (1 ') after and the step (3'), the following additional steps 'process according to any one of claims 21-24 comprising: (2)
　(2') after the condensation adding a quenching agent to the reaction solution.
[Requested item 26]
　Quenching agent used in step (2 ') The production method according to alcohols, claim 25 is at least one selected from phenols and amines.
[Requested item 27]
　To the reaction solution before the step (1 ') after and the step (4'), mixtures of carboxylic acids and organic bases, adding an inorganic acid or an amine, in the step (3 '), as a sulfurizing agent 5- [ (N, N-dimethylaminomethylidene) amino]-3H-1,2,4-dithiazole-3 process according to any of claims 21 to 26-thione is added to the reaction solution.
[Requested item 28]
　The amount of the basic nitrogen atoms of the organic base has the method according to claim 27 which is 1 to 2 mol with respect to the carboxy group 1 mol having a carboxylic acid.
[Requested item 29]
　Sulfurizing agent used in the step (3 ') is, 3H-1,2-benzodithiol-3-one 1,1-dioxide, 3H-1,2-benzodithiole-3-one, phenylacetyl disulfide, tetraethyl thiuram disulfide, dipentamethylenethiuram tetrasulfide, is at least one selected phenyl-3H-1,2,4-dithiazole-3-one, from 3-amino-1,2,4-dithiazole-5-thione and sulfur the process according to any one of claims 21 to 26.
[Requested item 30]
　Oxidizing agent used in the step (3 ') is iodine, (1S) - (+) - (10- camphorsulfonic sulfonyl) oxaziridine, tert- butyl hydroperoxide, 2-butanone peroxide, 1,1-dihydroperoxy cyclododecane , bis (trimethylsilyl) peroxide and m- chloro least one manufacturing method according to any one of claims 21 to 26 is selected from peracetic acid.
[Requested item 31]
　Temporary custody group hydroxyl groups, the manufacturing method according to any of claims 21 to 30, which is a dimethoxytrityl group or monomethoxytrityl group.
[Requested item 32]
　Nonpolar solvent, the production method according to halogenated solvents, any one of claims 21-31 is at least one selected from aromatic solvents, ester solvents and aliphatic solvents.
[Requested item 33]
　Acid used in step (4 ') is, trifluoroacetic acid, dichloroacetic acid, trifluoromethanesulfonic acid, trichloroacetic acid, methanesulfonic acid, hydrochloric acid, according to claim at least one selected from acetic acid and p- toluenesulfonic acid 21 the process according to any one of 1-32.
[Requested item 34]
　Not removed under acidic conditions, a protective group removable under basic conditions, aliphatic hydrocarbon radical having either a straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms, or branched one or more at least one a, and the production method according to any of claims 21 to 33 total carbon atoms with organic groups is 14 or more 300 or less.
[Requested item 35]
　Step (6 ') is, oligonucleotides (e' adding a polar solvent to a reaction solution containing a) any of claims 21 to 34, which is purifying the solid-liquid separation of oligonucleotides (e ') the method according to.
[Requested item 36]
　The process according to the polar solvent used in step (6 ') is any of claims 21 to 35, which is a nitrile solvent.
[Requested item 37]
　Before the step (4 ') after, and step (6'), further the following steps (5 ') The process according to any one of claims 21 to 36 including a:
　(5') oligonucleotides ( a step of neutralization by adding a base to the reaction solution containing e ').
[Requested item 38]
　Base used in step (5 ') is, pyridine, 2,4,6-trimethylpyridine, benzimidazole, 1,2,4-triazole, N- phenylimidazole, 2-amino-4,6-dimethylpyrimidine, 1 , 10-phenanthroline, imidazole, N- methylimidazole, at least 2-chloro-benzimidazole, selected 2-bromo-benzimidazole, 2-methylimidazole, 2-phenylbenzimidazole, N- phenyl-benzimidazole and 5-nitro-benzimidazole the process according to claim 37 which is one.
[Requested item 39]
　'After further following steps (7 step (6)') The process according to any one of claims 21 to 38 including a:
　(7 ') protecting groups of the resulting oligonucleotides were all removed after, isolating the unprotected oligonucleotide.
[Requested item 40]
　Step (4 ') Pauses or effect removal of the protecting group in the presence of a cation scavenger, or any of claims 21 to 39, the addition of a cation scavenger to the reaction solution after removal reaction of the temporary protecting group of the method according to.
[Requested item 41]
　Cation scavenger, pyrrole, 2-methylpyrrole, 3-methylpyrrole, 2,3-dimethylpyrrole, 2,4-dimethylpyrrole, indole, 3-methylindole, 4-methylindole, 5-methylindole, 6 methylindole, 7-methylindole, 5,6-dimethyl indole, 6,7-dimethyl indole, 2-methylfuran, 2,3-dimethyl furan, 2-methyl-3- (methylthio) furan, and selected from menthofuran the method according to claim 40 is at least one.
[Requested item 42]
　Formula (a-II):
[Chemical formula 1]

wherein,
　m represents an integer of 0 or more;
　the m Base are independently protected shows a nucleic acid base which may
　have; m + 1 pieces of X each independently represents a hydrogen atom, a halogen atom, optionally protected hydroxyl group or the 2-position a divalent organic group bonded to a carbon atom and 4 carbon atoms,;
　m-number of R 10 is, each independently represent an oxygen atom or a sulfur atom;
　m-number of R p1 are each independently a protecting group of a phosphate group;
　L represents a single bond or the formula (a1) or (a1 ') :
[Chemical formula 2]

(wherein,
　* indicates the bonding position to
　Y; ** indicates the bonding position to the oxygen
　atom; R 1 and R 2 are each independently, C 1-22 hydrocarbons It represents a hydrogen
　group; L 1 may be substituted divalent C 1-22It represents a hydrocarbon
　group; L 2 is either a single bond, or C *** (= O) N (R 3 ) -R 4 -N (R 5 ) **** (wherein *** is L 1 represents a bonding position with, **** denotes the bonding position to C = O, R 4 is C 1-22 represents an alkylene group, R 3 and R 5 are each independently Te, a hydrogen atom or a C 1-22 represents an alkyl group, or R 3 and R 5 have the together represent a group represented by the ring a may form.). A group represented by);
　Y is a single bond, an oxygen atom or NR (R, represents a hydrogen atom, an alkyl group or an aralkyl group); the. And
　Z has the formula (a2), the formula ( a2 ') or formula (a2 "):
[Chemical formula 3]

[wherein,
　* indicates the bonding
　position; R 6 is a hydrogen atom, or R b is, when a group represented by the following formula (a3), the R of the ring A or ring B 6 is R 8 single bond or together with It shows a -O-, ring a or ring B and ring C may form a fused ring with;
　k is from 1 to an integer of 4;
　k number of Q are each independently, -O -, - C (= O) -, - C (= O) O -, - OC (= O) -, - C (= O) NH- or -NHC (= O) - indicates;
　k number of R 7 are each independently an aliphatic hydrocarbon having either a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker, or branched one or more having at least one group, and total carbon number represents an organic group is 14 or more 300 or less;
　ring a and ring B are each independently, k pieces QR of 7 in addition to further halogen atoms, optionally substituted by a halogen atom C 1-6 alkyl group, and a C be substituted with halogen atoms 1-6 is selected from the group consisting of an alkoxy group It may have a substituent;
　R a represents a hydrogen atom; and
　R b is a hydrogen atom or formula (a3):
[Chemical formula 4]

(wherein,
　* indicates the bonding position;
　j is 0 to an integer of 4;
　j-number of Q are each independently the a be synonymous;
　j-number of R 9 is each independently a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker, or branched chain having at least one aliphatic hydrocarbon group having 1 or more, and the total number of carbon atoms represents an organic group of 14 to 300;
　R 8 is either a hydrogen atom, or a ring a or ring R of B 6 shows a single bond or -O- together with ring a or ring B and ring C may form a fused ring with; and
　ring C, j-number of QR 9 in addition to the Te further halogen atoms, which may be C-substituted by a halogen atom 1-6 alkyl group, and Even a C substituted by a halogen atom 1-6 or a group represented by which may have a substituent selected from the group consisting of an alkoxy group.), Or
　R aAnd R b is, form an oxo group together. It shows a group represented by.
　R 11 represents a methyl group, R 12 and R 13 are each independently C 1-5 represents an alkyl group, or R 11 and R 12 in the together, the carbon and nitrogen atoms they are attached it may form a nitrogen-containing hydrocarbon ring of 5-membered or 6-membered together. ]
Nucleoside or oligonucleotide represented by.
[Requested item 43]
　R p1 is, -CH 2 CH 2 WG (wherein, WG represents an electron-withdrawing group.) Nucleoside or oligonucleotide according to claim 42 is a group represented by.
[Requested item 44]
　R 11 is a methyl group, R 12 and R 13 are each independently C 1-5 nucleoside or oligonucleotide according to claim 42 or 43 is an alkyl group.
[Requested item 45]
　Nucleosides according to any one of claims 42 ~ 44 m is 0.
[Requested item 46]
　Formula (a-IV):
[Chemical Formula 5]

[wherein,
　m represents an integer of 0 or
　more; m + 1 pieces of Base is independently protected shows a nucleic acid base which may
　have; m + 1 pieces of X each independently represents a hydrogen atom, a halogen atom, optionally protected hydroxyl group or the 2-position a divalent organic group bonded to a carbon atom and 4 carbon atoms,;
　m-number of R 10 is, each independently represent an oxygen atom or a sulfur atom;
　m-number of R p1 are each independently a protecting group of a phosphate group;
　L represents a single bond or the formula (a1) or (a1 ') :
[formula 6]

(wherein,
　* indicates the bonding position to
　Y; ** indicates the bonding position to the oxygen
　bond; R 1 and R 2 are each independently, C 1-22 hydrocarbons It represents a hydrogen
　group; L 1 may be substituted divalent C 1-22It represents a hydrocarbon
　group; L 2 is either a single bond, or C *** (= O) N (R 3 ) -R 4 -N (R 5 ) **** (wherein *** is L 1 represents a bonding position with, **** denotes the bonding position to C = O, R 4 is C 1-22 represents an alkylene group, R 3 and R 5 are each independently Te, a hydrogen atom or a C 1-22 represents an alkyl group, or R 3 and R 5 have the together represent a group represented by the ring a may form.). Represented by a group in);
　. Y represents a single bond, an oxygen atom or NR, (R represents a represents a hydrogen atom, an alkyl group or an aralkyl group); and
　Z 'has the formula (a2 "):
[Chemical formula 7]

wherein,
　* represents a bonding
　position; R 6Is a hydrogen atom, or R b is, when a group represented by the following formula (a3) is, R 8 represents a single bond or -O- together with ring B and ring may form a fused ring together with C;
　k is from 1 to an integer of 4;
　k number of Q are each independently, -O -, - C (= O) -, - C (= O) O -, - OC ( = O) -, - C (= O) NH- or -NHC (= O) - are shown;
　k-number of R 7 are each independently a straight 10 or more carbon atoms or a hydrocarbon radical chain aliphatic hydrocarbon group is linked via a single bond or a linker, or branched chain at least one has an aliphatic hydrocarbon group having 1 or more, and the total carbon number of 14 indicates a is an organic group 300 inclusive;
　ring B, k-number of QR 7 in addition to further halogen atoms, substituted with a halogen atom Which may be C 1-6 alkyl groups, and substituted with a halogen atom which may C 1-6 may have a substituent group selected from the group consisting of an alkoxy group;
　R a is a hydrogen atom shown; and
　R b is a hydrogen atom or the formula, (a3):
[formula 8]

(Wherein,
　* indicates the bonding position;
　j is 0 to an integer of 4;
　j-number of Q are each independently a as defined above;
　j-number of R 9 is each independently Te, at least one organic aliphatic hydrocarbon radical having either a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker, or branched one or more of and, and the total number of carbon atoms is an organic group of 14 to 300;
　R 8 is either a hydrogen atom, or R 6 represents a single bond or -O- together with ring B and ring It may form a fused ring together with C; and
　ring C, j-number of QR 9 in addition to further halogen atom, a C substituted with halogen atom 1-6 alkyl group, and a halogen atom unsubstituted or substituted C have 1-6 or consisting alkoxy group Or a group represented by the selected may have a substituent.), Or
　R a and R b are taken together to form an oxo group. It shows a group represented by. ]
In represented nucleoside or oligonucleotide.
[Requested item 47]
　R p1 is, -CH 2 CH 2 WG (wherein, WG represents an electron-withdrawing group.) Nucleoside or oligonucleotide according to claim 46 is a group represented by.
[Requested item 48]
　Nucleoside of claim 46 or 47 m is 0.
[Requested item 49]
　Formula (I):
[Chemical Formula 9]

[wherein,
　m represents an integer of 0 or more;
　m + 1 pieces of Base are each independently a protected or unprotected nucleobase;
　m + 1 pieces of X is each, independently, a hydrogen atom, a halogen atom, optionally protected hydroxyl group or 2-position represents a divalent organic group bonded to a carbon atom and 4 carbon atoms,;
　m-number of R 10 is independently to, an oxygen atom or a sulfur atom;
　m-number of R p1 are each independently a protecting group of a phosphate group;
　R n1 and R n2 one is a hydrogen atom, the remaining one has the formula (II):
[Chemical formula 10]

{wherein,
　* indicates the bonding
　position; R 10 represents an oxygen atom or a sulfur
　atom; R p1 is a protecting group of a phosphate
　group; L n1 of the organic group the shows;
　L represents a single bond or the formula (a1) or (a1 '):
[Formula 11]

(wherein,
　* indicates the bonding position to
　Y; ** indicates the bonding position to the oxygen
　atom; R 1 and R 2 are each independently, C 1-22 a hydrocarbon
　group; L 1 is C divalent optionally substituted 1-22 represents a hydrocarbon
　group; L 2 is a single bond or shows, or C *** (= O) N (R 3 ) -R 4 -N (R 5 ) **** (wherein, *** is, L 1 represents a bonding position with the * *** indicates the bonding position to C = O, R 4 is C 1-22 represents an alkylene group, R 3 and R 5They are each independently a hydrogen atom or a C 1-22 represents an alkyl group, or R 3 and R 5 are taken together, may form a ring. ) A group represented by. A group represented by);
　Y is a single bond, an oxygen atom or NR (R, represents a hydrogen atom, an alkyl group or an aralkyl group); the. And
　Z has the formula (a2), the formula ( a2 ') or formula (a2 "):
[formula 12]

wherein,
　* represents a bonding
　position; R 6 is a hydrogen atom, or R b is represented by the following formula (a3) If a group, R a ring a or ring B 6 is, R 8 represents a single bond or -O- together with form a fused ring together with ring a or ring B and ring C ; even better
　is k, 1 ~ represents an integer of 4;
　k number of Q are each independently, -O -, - C (= O) -, - C (= O) O -, - OC (= O) -, - C (= O) NH- or -NHC (= O) - indicates;
　k-number of R 7Are each independently, an aliphatic hydrocarbon radical having either a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker, or branched one or more the has at least one, and the total carbon number of an organic group is 14 or more 300 or less;
　ring a and ring B are each independently, k-number of QR 7 in addition to further halogen atom, a halogen atom in an optionally substituted C 1-6 alkyl group, and a halogen atom in the optionally substituted C 1-6 may have a substituent group selected from the group consisting of an alkoxy group;
　R a is It represents a hydrogen atom; and
　R b is a hydrogen atom or formula (a3):
[formula 13]

(wherein,
　* indicates the bonding position;
　j represents an integer of 0 ~ 4;
　j-number of Q are each independently a as defined above;
　j-number of R 9 are each independently an aliphatic having either a hydrocarbon group which is straight-chain aliphatic hydrocarbon group having 10 or more carbon atoms linked through a single bond or a linker, or branched one or more carbide having at least one hydrogen group, and total carbon number represents an organic group of 14 to 300;
　R 8 is either a hydrogen atom, or R of ring A or ring B 6 together with by a single bond or -O-, form a fused ring together with ring A or ring B and ring C It is good; and
　ring C, j-number of QR 9 in addition to further halogen atom, a C substituted with halogen atom 1-6 alkyl group, and may be substituted by a halogen atom C 1- 6 a substituent selected from the group consisting of an alkoxy group may have. ) On whether a group represented, or
　R a and R b are, form an oxo group together. It shows a group represented by. A group represented by}. ]
Or oligonucleotide represented by.
[Requested item 50]
　R n1 is a group is represented by the formula (II), R n2 or oligonucleotide according to claim 49 is a hydrogen atom.
[Requested item 51]
　L n1 or oligonucleotide according to claim 49 or 50 is an ethylene group.
[Requested item 52]
　Nucleotides according to any one of claims 49 ~ 51 m is 0.