1. A precipitation promoter for precipitating an organic compound protected by an organic group having one or more aliphatic hydrocarbon groups having not less 5 than 10 carbon atoms in a solvent, which has one or more linear aliphatic hydrocarbon groups having not less than 10 carbon atoms, wherein the aliphatic hydrocarbon group in the precipitation promoter has not less than 20 carbon atoms in total. 10

2. The precipitation promoter according to claim 1, wherein the linear aliphatic hydrocarbon group having not less than 10 carbon atoms in the precipitation promoter is a group selected from a linear C10-40 alkyl group and a linear C10-40 alkenyl group. 15

3. The precipitation promoter according to claim 1 or 2, wherein the aliphatic hydrocarbon group having not less than 10 carbon atoms of the organic group is linear. 20 4. The precipitation promoter according to claim 1 or 2, wherein the aliphatic hydrocarbon group having not less than 10 carbon atoms of the organic group is a group selected from a linear C10-40 alkyl group and a linear C10-40 alkenyl group. 25 5. The precipitation promoter according to any one of claims 1 to 4, which is (1) an organic compound having one or more structures represented by the formula (G): wherein each R1 30 is independently a linear C10-40 alkyl group; each X1 is independently a single bond, -O-, -C(=O)-, - C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-; ring A1 is an optionally substituted C3-14 hydrocarbon 130 ring; and n is an integer of 1 - 4, or (2) optionally substituted C1-10 alkane having one or more linear C10-40 alkyl groups via a group selected from the group consisting of -O-, -C(=O)-, -C(=O)O-, -OC(=5 O)-, -C(=O)NH- and - NHC(=O)-.

6. The precipitation promoter according to any one of claims 1 to 4, which is (1) a compound represented by the formula (I): 10 wherein each R1 is independently a linear C10-40 alkyl group; each X1 is independently a single bond, -O-, -C(=O)-, - C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-; ring A1 and ring B1 15 are each independently an optionally substituted C3-14 hydrocarbon ring; each X2 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, - (CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q-, -(CH2)p-OC(=O)- (CH2)q-, -(CH2)p-C(=O)NH-(CH2)q- or -(CH2)p-NHC(=O)-(CH2)q- (p and 20 q are each independently an integer of 0 - 3); X3 is a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-, -(CH2)r- C(=O)O-, -(CH2)r-OC(=O)-, -(CH2)r-C(=O)NH- or -(CH2)r-NHC(=O)- (r is an integer of 0 - 3); R2 is a hydrogen atom, an optionally substituted C3-8 25 cycloalkyl group, an optionally substituted C1-6 alkyl group, an optionally substituted C2-6 alkenyl group, an optionally substituted C2-6 alkynyl group, an optionally substituted C6-14 aryl group, an optionally substituted monocyclic heterocyclic group or a tri(C1-6 alkyl)silyl group; 30 n and m are each independently an integer of 1 - 4; m’ is 0 or 1 when m is 1, and 1 when m is 2, 3 or 4, or (2) optionally substituted C1-10 alkane having one or more linear C10-40 alkyl groups via a group selected from the group 131 consisting of -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH- and - NHC(=O)-.

7. The precipitation promoter according to claim 6, wherein R2 is a hydrogen atom, an optionally substituted 5 C3-8 cycloalkyl group, an optionally substituted C1-6 alkyl group, an optionally substituted C2-6 alkenyl group, an optionally substituted C2-6 alkynyl group, an optionally substituted C6-14 aryl group or an optionally substituted monocyclic heterocyclic group. 10

8. The precipitation promoter according to any one of claims 1 to 4, which is a compound represented by the formula (II): wherein each R1 15 is independently a linear C10-40 alkyl group; each X1 is independently a single bond, -O-, -C(=O)-, - C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-; ring A2 and ring B1 are each independently an optionally substituted C3-14 hydrocarbon ring; each X2 20 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, - (CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q-, -(CH2)p-OC(=O)- (CH2)q-, -(CH2)p-C(=O)NH-(CH2)q- or -(CH2)p-NHC(=O)-(CH2)q- (p and q are each independently an integer of 0 - 3); X3 is a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-, -(CH2)r- 25 C(=O)O-, -(CH2)r-OC(=O)-, -(CH2)r-C(=O)NH- or -(CH2)r-NHC(=O)- (r is an integer of 0 - 3); R2 is a hydrogen atom, an optionally substituted C3-8 cycloalkyl group, an optionally substituted C1-6 alkyl group, an optionally substituted C2-6 alkenyl group, an optionally 30 substituted C2-6 alkynyl group, an optionally substituted C6-14 aryl group, an optionally substituted monocyclic heterocyclic 132 group or a tri(C1-6 alkyl)silyl group; m is an integer of 1 to 4; and m’ is 0 or 1 when m is 1, and 1 when m is 2, 3 or 4.

9. The precipitation promoter according to claim 8, wherein R2 5 is a hydrogen atom, an optionally substituted C3-8 cycloalkyl group, an optionally substituted C1-6 alkyl group, an optionally substituted C2-6 alkenyl group, an optionally substituted C2-6 alkynyl group, an optionally substituted C6-14 aryl group or an 10 optionally substituted monocyclic heterocycle.

10. The precipitation promoter according to any one of claims 5 to 9, wherein the C3-14 hydrocarbon ring is selected from a benzene ring and a cyclohexane ring. 15

11. The precipitation promoter according to any one of claims 1 to 10, wherein the organic group is a group represented by the formula (III): 20 wherein ** shows a bonding position to a group to be protected; L is a single bond, or a group represented by the formula (a1) or (a1’): 25 wherein * shows the bonding position to Y; ** is as defined above; R8 and R9 are each independently a C1-22 hydrocarbon group; 30 L1 is a divalent C1-22 hydrocarbon group; and 133 L2 is a single bond, or a group represented by **C(=O)N(R2’)-R1’- N(R3)*** wherein ** shows the bonding position to L1, *** shows the bonding position to C=O, R1’ is a C1-22 alkylene group, and R2’ and R3 are each independently a hydrogen atom or a C1-22 alkyl group, or R2’ and R3 are optionally joined 5 to form a ring, Y is a single bond, an oxygen atom or NR wherein R is a hydrogen atom, an alkyl group or an aralkyl group, and Z is a group represented by the formula (a2), the formula (a2’) or the formula (a2”): 10 wherein * shows a bonding position; R4 is a hydrogen atom, or when Rb is a group represented by the following formula (a3), optionally joined with R6 of ring C to 15 show a single bond or -O- and to form a fused ring together with ring A or ring B and ring C; Q in the number of k are each independently -O-, -C(=O)-, - C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-; R5 in the number of k are each independently a hydrocarbon 20 group bonded via a single bond or a linker to an aliphatic hydrocarbon group having not less than 10 carbon atoms; k is an integer of 1 to 4; ring A and ring B are each independently optionally further have, in addition to QR5 in the number of k, substituent(s) 25 selected from the group consisting of a halogen atom, a C1-6 alkyl group optionally substituted by a halogen atom, and a C1-6 alkoxy group optionally substituted by a halogen atom; Ra is a hydrogen atom, or a phenyl group optionally substituted by a halogen atom; and 30 Rb is a hydrogen atom, or a group represented by the formula 134 (a3): wherein * shows a bonding position; j is an integer of 0 to 4; Q in the number of j are each independently 5 as defined above; R7 in the number of j are each independently a hydrocarbon group bonded via a single bond or a linker to an aliphatic hydrocarbon group having not less than 10 carbon atoms; R6 is a hydrogen atom, or optionally joined with R4 of ring A 10 or ring B to show a single bond or -O- and to form a fused ring together with ring A or ring B and ring C; and ring C optionally further has, in addition to OR7 in the number of j, substituent(s) selected from the group consisting of a halogen atom, a C1-6 alkyl group optionally substituted by a 15 halogen atom, and a C1-6 alkoxy group optionally substituted by a halogen atom, or Ra and Rb are joined to form an oxo group.

12. The precipitation promoter according to claim 11, wherein Z 20 is a group represented by the formula (a2) or the formula (a2’), the fused ring is a fluorene ring or a xanthene ring, R5 in the number of k are each independently a hydrocarbon group bonded via a single bond or a linker to a linear aliphatic hydrocarbon group having not less than 10 carbon atoms, R7 in the number of 25 j are each independently a hydrocarbon group bonded via a single bond or a linker to a linear aliphatic hydrocarbon group having not less than 10 carbon atoms, and Ra is a hydrogen atom, or Ra and Rb are joined to form an oxo group. 30 13. The precipitation promoter according to claim 11 or 12, wherein L is a group represented by the formula (a1), L1 is a divalent C1-22 hydrocarbon group, and L2 is a single bond. 135

14. The precipitation promoter according to claim 11 or 12, wherein L is a group represented by the formula (a1’), L1 is a phenylene group, and L2 is a single bond. 5

15. The precipitation promoter according to any one of claims 11 to 14, wherein Y is an oxygen atom.

16. The precipitation promoter according to any one of claims 10 11 to 15, wherein Z is a group represented by the formula (a2), and R4 is a hydrogen atom.

17. The precipitation promoter according to any one of claims 11 to 16, wherein Z is a group represented by the formula (a2), 15 and Ra and Rb are each a hydrogen atom.

18. The precipitation promoter according to claim 11 or 12, wherein L and Y are each a single bond, Z is a group represented by the formula (a2), R4 is a hydrogen atom, and Ra 20 and Rb are joined to form an oxo group.

19. The precipitation promoter according to any one of claims 1 to 18, wherein the organic compound protected by the organic group is nucleoside, nucleotide or oligonucleotide optionally 25 further protected by a protecting group used in nucleic acid synthesis, or amino acid or peptide optionally further protected by a protecting group used in peptide synthesis.

20. The precipitation promoter according to any one of claims 1 30 to 18, wherein the organic compound protected by the organic group is nucleoside, nucleotide or oligonucleotide optionally further protected by a protecting group used in nucleic acid synthesis. 35 21. The precipitation promoter according to any one of claims 1 136 to 18, wherein the organic compound protected by the organic group is nucleoside or oligonucleotide wherein at least one group selected from an amino group and an imino group of a nucleic acid base, 2’- and 3’-hydroxy groups of a ribose residue, and 3’-hydroxy group of a deoxyribose 5 residue is protected by the organic group, and other group is optionally further protected by a protecting group used in nucleic acid synthesis. 10 22. The precipitation promoter according to any one of claims 1 to 21, wherein the solvent comprises a polar solvent.

23. The precipitation promoter according to claim 22, wherein the solvent comprising a polar solvent is a mixed solvent of a 15 polar solvent and a nonpolar solvent.

24. The precipitation promoter according to claim 22 or 23, wherein the polar solvent is acetonitrile. 20 25. The precipitation promoter according to any one of claims 1 to 24, which is used at not less than 0.1 molar equivalent relative to the organic compound protected by the organic group to precipitate the organic compound protected by the organic group. 25

26. A precipitation mixture comprising the precipitation promoter according to any one of claims 1 to 25, and an organic compound protected by an organic group having one or more aliphatic hydrocarbon groups having not less than 10 carbon 30 atoms.

27. The precipitation mixture according to claim 26, wherein the aliphatic hydrocarbon group having not less than 10 carbon atoms of the organic group is linear. 35 137

28. A method of precipitating an organic compound protected by an organic group having one or more aliphatic hydrocarbon groups having not less than 10 carbon atoms in a solvent, by using the precipitation promoter according to any one of claims 5 1 to 25.

29. The method according to claim 28, wherein the solvent comprises a polar solvent. 10 30. The method according to claim 29, wherein the polar solvent is acetonitrile.

31. The method according to any one of claims 28 to 30, wherein the aliphatic hydrocarbon group having not less than 10 carbon 15 atoms of the organic group is linear

32. A production method of an oligonucleotide, comprising a step of adding a polar solvent to a reaction solution comprising an oligonucleotide wherein at least one group is 20 protected by an organic group having one or more aliphatic hydrocarbon groups having not less than 10 carbon atoms, and other group is optionally further protected by a protecting group used in nucleic acid synthesis, and the precipitation promoter according to any one of claims 1 to 25 in a nonpolar 25 solvent, and separating a precipitate mixture comprising the oligonucleotide and the precipitation promoter from the reaction solution.

33. The production method according to claim 32, wherein the 30 polar solvent is acetonitrile.

34. The production method according to claim 32 or 33, wherein the aliphatic hydrocarbon group having not less than 10 carbon atoms of the organic group is linear. 35 138

35. The production method according to any one of claims 32 to 34, which is performed by a phosphoramidite method.

36. A production method of an oligonucleotide, which includes one repeat of production cycle comprising the 5 following steps (1) - (3), or plural repeats thereof by a phosphoramidite method, which comprises the following step (4) in the first cycle, the following step (5) in each cycle, and the following step (6) in each cycle except the final cycle: 10 (1) a step of obtaining a reaction solution comprising a free- 5’-hydroxy-group form by adding an acid to a reaction solution comprising a nucleoside or oligonucleotide wherein at least one group selected from an amino group and an imino group of a nucleic acid base, 2’- and 3’-hydroxy groups of a ribose 15 residue, and 3’-hydroxy group of a deoxyribose residue is protected by an organic group having one or more aliphatic hydrocarbon groups having not less than 10 carbon atoms, a 5’- hydroxy group is protected by a temporary protecting group removable under acidic conditions, and other group is 20 optionally further protected by a protecting group used in nucleic acid synthesis in a nonpolar solvent, to deprotect the temporary protecting group of the 5’-hydroxy group, and neutralizing same with a base; (2) a step of obtaining a reaction solution comprising a 25 phosphite triester form, by adding nucleoside or oligonucleotide wherein a 3’-hydroxy group is phosphoramidited, a 5’-hydroxy group is protected by a temporary protecting group removable under acidic conditions, and other group is optionally further protected by a protecting group used in 30 nucleic acid synthesis to the reaction solution comprising the free-5’-hydroxy-group form in a nonpolar solvent; (3) a step of obtaining a reaction solution comprising an oligonucleotide wherein at least one group selected from an amino group and an imino group of a nucleic acid base, 2’- and 35 3’-hydroxy groups of a ribose residue, and 3’-hydroxy group of 139 a deoxyribose residue is protected by an organic group having one or more aliphatic hydrocarbon groups having not less than 10 carbon atoms, a 5’-hydroxy group is protected by a temporary protecting group removable under acidic conditions, and other group is optionally further protected by 5 a protecting group used in nucleic acid synthesis, by adding an oxidizing agent or a sulfurizing agent to the reaction solution comprising the phosphite triester form in a nonpolar solvent; (4) a step of adding the precipitation promoter according to 10 any one of claims 1 to 25 to the reaction solution at any of before step (1), between steps (1) and (2), between steps (2) and (3) and after step (3); (5) a step of separating a precipitation mixture comprising the free-5’-hydroxy-group form, the phosphite triester form or the 15 oligonucleotide, and the precipitation promoter from the reaction solution by adding a polar solvent to the reaction solution comprising the precipitation promoter at after step (4), and any of between steps (1) and (2), between steps (2) and (3) and after step (3); 20 (6) a step of adding a nonpolar solvent to the precipitation mixture obtained in step (5) to give a reaction solution.

37. The production method according to claim 36, wherein the polar solvent is acetonitrile. 25

38. The production method according to claim 36 or 37, wherein the aliphatic hydrocarbon group having not less than 10 carbon atoms of the organic group is linear. 30 39. The method according to any one of claims 36 to 38, wherein the steps (5) and (6) are performed after step (3).

40. The method according to any one of claims 36 to 39, further comprising the following step (7): 35 (7) a step of removing all protecting groups of the 140 oligonucleotide and isolating the oligonucleotide.

41. The method according to any one of claims 36 to 40, wherein the non-polar solvent is a solvent selected from the group consisting of a halogenated solvent, an aromatic 5 solvent, an ester solvent, an aliphatic solvent, and a combination thereof.

Title of the Invention: PRECIPITATION ACCELERATOR AND
PRECIPITATION METHOD IN WHICH SAME IS USED
[Technical Field]
5 [0001]
The present invention relates to a precipitation promoter
useful for precipitating an organic compound (e.g.,
oligonucleotide, peptide etc.) in a solution, and a
precipitation method using same.
10 [Background Art]
[0002]
The synthesis method of oligonucleotide includes a
phosphate triester method, an H-phosphonate method, a
phosphoramidite method and the like, and solid phase synthesis
15 (solid phase method) using a phosphoramidite method is most
widely used at present. As a synthesis method of peptide,
moreover, solid phase synthesis is widely used. The solid
phase method is advantageous from the aspect of speed, since
process has been optimized and automation has progressed.
20 However, it is associated with defects in that scaling-up is
limited due to facility restriction, reagents and starting
materials are used in excess, and confirmation of the progress
status of the reaction in an intermediate step, analysis of
intermediate structure and the like are difficult. On the
25 other hand, synthesis methods of oligonucleotide and peptide by
a liquid phase method have also been studied. However, since
the operation is complicated and the yield is low, a largescale,
rapid synthesis of long oligonucleotide and peptide is
difficult.
30 [0003]
In recent years, in an attempt to solve the respective
defects of the solid phase method and the liquid phase method,
there have been reported, in the liquid phase method, a
synthesis method of oligonucleotide, which uses a nucleoside
35 protected by an organic group having one or more long chain
3
aliphatic hydrocarbon groups (pseudo solid phase protecting
group) (patent documents 1 and 2), and a synthesis method of
peptide, which uses a peptide having a pseudo solid phase
protecting group (patent documents 3, 4 and 5). In these
synthesis methods, for example, the object product 5 is recovered
by dissolving oligonucleotide or peptide having a hydrophobic
pseudo solid phase protecting group in a non-polar solvent, and
adding a polar solvent to allow for precipitation of these
compounds from a mixed solvent of the non-polar solvent and the
10 polar solvent (solid-liquid separation).
[Document List]
[Patent documents]
[0004]
patent document 1: WO 2012/157723
15 patent document 2: WO 2014/189142
patent document 3: WO 2010/104169
patent document 4: WO 2010/113939
patent document 5: WO 2011/078295
[SUMMARY OF THE INVENTION]
20 [Problems to be Solved by the Invention]
[0005]
In liquid phase synthesis of an organic compound such as
oligonucleotide, peptide and the like, which uses such pseudo
solid phase protecting group, more efficient precipitation of
25 these compounds in a solvent to improve the recovery rate of
the compounds has been desired.
[0006]
The present invention has been made taking note of the
above-mentioned situation, and aims to improve a recovery rate
30 of an organic compound having a pseudo solid phase protecting
group, when the organic compound is precipitated in a solvent.
[Means of Solving the Problems]
[0007]
The present inventors have conducted intensive studies in
35 an attempt to achieve the aforementioned object and found that
4
precipitation of an organic compound having a hydrophobic group
or pseudo solid phase protecting group can be promoted by using
a particular precipitation promoter, and the recovery rate and
the can be improved. The present invention based on these
findings is as 5 described below.
[0008]
[1] A precipitation promoter for precipitating an organic
compound protected by an organic group having one or more
aliphatic hydrocarbon groups having not less than 10 carbon
10 atoms in a solvent, which has one or more linear aliphatic
hydrocarbon groups having not less than 10 carbon atoms,
wherein the aforementioned aliphatic hydrocarbon group in the
precipitation promoter has not less than 20 carbon atoms in
total.
15 [2] The precipitation promoter of the aforementioned [1],
wherein the linear aliphatic hydrocarbon group having not less
than 10 carbon atoms in the precipitation promoter is a group
selected from a linear C10-40 alkyl group and a linear C10-40
alkenyl group.
20 [3] The precipitation promoter of the aforementioned [1] or [2],
wherein the aliphatic hydrocarbon group having not less than 10
carbon atoms of the organic group is linear.
[4] The precipitation promoter of the aforementioned [1] or [2],
wherein the aliphatic hydrocarbon group having not less than 10
25 carbon atoms of the organic group is a group selected from a
linear C10-40 alkyl group and a linear C10-40 alkenyl group.
[5] The precipitation promoter of any one of the aforementioned
[1] - [4], which is (1) an organic compound having one or more
structures represented by the formula (G):
30 [0009]
[0010]
wherein
5
each R1 is independently a linear C10-40 alkyl group;
each X1 is independently a single bond, -O-, -C(=O)-, -
C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
ring A1 is an optionally substituted C3-14 hydrocarbon
5 ring; and
n is an integer of 1 - 4, or
(2) optionally substituted C1-10 alkane having one or more
linear C10-40 alkyl groups via a group selected from the group
consisting of -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH- and -
10 NHC(=O)-.
[6] The precipitation promoter of any one of the aforementioned
[1] - [4], which is (1) a compound represented by the formula
(I):
[0011]
15
[0012]
wherein
each R1 is independently a linear C10-40 alkyl group;
each X1 is independently a single bond, -O-, -C(=O)-, -
20 C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
ring A1 and ring B1 are each independently an optionally
substituted C3-14 hydrocarbon ring;
each X2 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q-, -(CH2)p-OC(=O)-
25 (CH2)q-, -(CH2)p-C(=O)NH-(CH2)q- or -(CH2)p-NHC(=O)-(CH2)q- (p and
q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-, -(CH2)r-
C(=O)O-, -(CH2)r-OC(=O)-, -(CH2)r-C(=O)NH- or -(CH2)r-NHC(=O)- (r
is an integer of 0 - 3);
R2 30 is a hydrogen atom, an optionally substituted C3-8
cycloalkyl group, an optionally substituted C1-6 alkyl group, an
optionally substituted C2-6 alkenyl group, an optionally
substituted C2-6 alkynyl group, an optionally substituted C6-14
6
aryl group, an optionally substituted monocyclic heterocyclic
group or a tri(C1-6 alkyl)silyl group;
n and m are each independently an integer of 1 - 4;
m’ is 0 or 1 when m is 1, and 1 when m is 2, 3 or 4, or
(2) optionally substituted C1-10 alkane 5 having one or more
linear C10-40 alkyl groups via a group selected from the group
consisting of -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH- and -
NHC(=O)-.
[7] The precipitation promoter of the aforementioned [6],
wherein R2 10 is a hydrogen atom, an optionally substituted C3-8
cycloalkyl group, an optionally substituted C1-6 alkyl group, an
optionally substituted C2-6 alkenyl group, an optionally
substituted C2-6 alkynyl group, an optionally substituted C6-14
aryl group or an optionally substituted monocyclic heterocyclic
15 group.
[8] The precipitation promoter of any one of the aforementioned
[1] - [4], which is a compound represented by the formula (II):
[0013]
20 [0014]
wherein
each R1 is independently a linear C10-40 alkyl group;
each X1 is independently a single bond, -O-, -C(=O)-, -
C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
ring A2 and ring B1 25 are each independently an optionally
substituted C3-14 hydrocarbon ring;
each X2 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q-, -(CH2)p-OC(=O)-
(CH2)q-, -(CH2)p-C(=O)NH-(CH2)q- or -(CH2)p-NHC(=O)-(CH2)q- (p and
30 q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-, -(CH2)r7
C(=O)O-, -(CH2)r-OC(=O)-, -(CH2)r-C(=O)NH- or -(CH2)r-NHC(=O)- (r
is an integer of 0 - 3);
R2 is a hydrogen atom, an optionally substituted C3-8
cycloalkyl group, an optionally substituted C1-6 alkyl group, an
optionally substituted C2-6 alkenyl group, 5 an optionally
substituted C2-6 alkynyl group, an optionally substituted C6-14
aryl group, an optionally substituted monocyclic heterocyclic
group or a tri(C1-6 alkyl)silyl group;
m is an integer of 1 to 4; and
10 m’ is 0 or 1 when m is 1, and 1 when m is 2, 3 or 4.
[9] The precipitation promoter of the aforementioned [8],
wherein R2 is a hydrogen atom, an optionally substituted C3-8
cycloalkyl group, an optionally substituted C1-6 alkyl group, an
optionally substituted C2-6 alkenyl group, an optionally
15 substituted C2-6 alkynyl group, an optionally substituted C6-14
aryl group or an optionally substituted monocyclic heterocycle.
[10] The precipitation promoter of any one of the
aforementioned [5] - [9], wherein the C3-14 hydrocarbon ring is
selected from a benzene ring and a cyclohexane ring.
20 [11] The precipitation promoter of any one of the
aforementioned [1] - [10], wherein the organic group is a group
represented by the formula (III):
[0015]
25
[0016]
wherein
** shows a bonding position to a group to be protected;
L is a single bond, or a group represented by the formula
30 (a1) or (a1’):
[0017]
8
[0018]
wherein
* shows the bonding position to Y;
** is 5 as defined above;
R8 and R9 are each independently a C1-22 hydrocarbon group;
L1 is a divalent C1-22 hydrocarbon group; and
L2 is a single bond, or a group represented by **C(=O)N(R2’)-R1’-
N(R3)*** wherein ** shows the bonding position to L1, *** shows
the bonding position to C=O, R1’ 10 is a C1-22 alkylene group, and
R2’ and R3 are each independently a hydrogen atom or a C1-22
alkyl group, or R2’ and R3 are optionally joined to form a ring,
Y is a single bond, an oxygen atom or NR wherein R is a
hydrogen atom, an alkyl group or an aralkyl group, and
15 Z is a group represented by the formula (a2), the formula (a2’)
or the formula (a2”):
[0019]
[0020]
20 wherein
* shows a bonding position;
R4 is a hydrogen atom, or when Rb is a group represented by the
following formula (a3), optionally joined with R6 of ring C to
show a single bond or -O- and to form a fused ring together
25 with ring A or ring B and ring C;
9
Q in the number of k are each independently -O-, -C(=O)-, -
C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
R5 in the number of k are each independently a hydrocarbon
group bonded via a single bond or a linker to an aliphatic
hydrocarbon group having not less than 5 10 carbon atoms;
k is an integer of 1 to 4;
ring A and ring B are each independently optionally further
have, in addition to QR5 in the number of k, substituent(s)
selected from the group consisting of a halogen atom, a C1-6
10 alkyl group optionally substituted by a halogen atom, and a C1-6
alkoxy group optionally substituted by a halogen atom;
Ra is a hydrogen atom, or a phenyl group optionally substituted
by a halogen atom; and
Rb is a hydrogen atom, or a group represented by the formula
15 (a3):
[0021]
[0022]
wherein * shows a bonding position;
20 j is an integer of 0 to 4;
Q in the number of j are each independently as defined above;
R7 in the number of j are each independently a hydrocarbon
group bonded via a single bond or a linker to an aliphatic
hydrocarbon group having not less than 10 carbon atoms;
R6 is a hydrogen atom, or optionally joined with R4 25 of ring A
or ring B to show a single bond or -O- and to form a fused ring
together with ring A or ring B and ring C; and
ring C optionally further has, in addition to OR7 in the number
of j, substituent(s) selected from the group consisting of a
30 halogen atom, a C1-6 alkyl group optionally substituted by a
halogen atom, and a C1-6 alkoxy group optionally substituted by
a halogen atom, or
10
Ra and Rb are joined to form an oxo group.
[12] The precipitation promoter of the aforementioned [11],
wherein Z is a group represented by the formula (a2) or the
formula (a2’), the fused ring is a fluorene ring or a xanthene
ring, R5 in the number of k are each 5 independently a
hydrocarbon group bonded via a single bond or a linker to a
linear aliphatic hydrocarbon group having not less than 10
carbon atoms, R7 in the number of j are each independently a
hydrocarbon group bonded via a single bond or a linker to a
10 linear aliphatic hydrocarbon group having not less than 10
carbon atoms, and Ra is a hydrogen atom, or Ra and Rb are joined
to form an oxo group.
[13] The precipitation promoter of the aforementioned [11] or
[12], wherein L is a group represented by the formula (a1), L1
15 is a divalent C1-22 hydrocarbon group, and L2 is a single bond.
[14] The precipitation promoter of the aforementioned [11] or
[12], wherein L is a group represented by the formula (a1’), L1
is a phenylene group, and L2 is a single bond.
[15] The precipitation promoter of any one of the
20 aforementioned [11] - [14], wherein Y is an oxygen atom.
[16] The precipitation promoter of any one of the
aforementioned [11] - [15], wherein Z is a group represented by
the formula (a2), and R4 is a hydrogen atom.
[17] The precipitation promoter of any one of the
25 aforementioned [11] - [16], wherein Z is a group represented by
the formula (a2), and Ra and Rb are each a hydrogen atom.
[18] The precipitation promoter of the aforementioned [11] or
[12], wherein L and Y are each a single bond, Z is a group
represented by the formula (a2), R4 is a hydrogen atom, and Ra
30 and Rb are joined to form an oxo group.
[19] The precipitation promoter of any one of the
aforementioned [1] - [18], wherein the organic compound
protected by the organic group is nucleoside, nucleotide or
oligonucleotide optionally further protected by a protecting
35 group used in nucleic acid synthesis, or amino acid or peptide
11
optionally further protected by a protecting group used in
peptide synthesis.
[20] The precipitation promoter of any one of the
aforementioned [1] - [18], wherein the organic compound
protected by the organic group is nucleoside, 5 nucleotide or
oligonucleotide optionally further protected by a protecting
group used in nucleic acid synthesis.
[21] The precipitation promoter of any one of the
aforementioned [1] - [18], wherein the organic compound
10 protected by the organic group is nucleoside or oligonucleotide
wherein at least one group selected from an amino group and an
imino group of a nucleic acid base, 2’- and 3’-hydroxy groups
of a ribose residue, and 3’-hydroxy group of a deoxyribose
residue is protected by the organic group, and other group is
15 optionally further protected by a protecting group used in
nucleic acid synthesis.
[22] The precipitation promoter of any one of the
aforementioned [1] - [21], wherein the solvent comprises a
polar solvent.
20 [23] The precipitation promoter of the aforementioned [22],
wherein the solvent comprising a polar solvent is a mixed
solvent of a polar solvent and a nonpolar solvent.
[24] The precipitation promoter of the aforementioned [22] or
[23], wherein the polar solvent is acetonitrile.
25 [25] The precipitation promoter of any one of the
aforementioned [1] - [24], which is used at not less than 0.1
molar equivalent relative to the organic compound protected by
the organic group to precipitate the organic compound protected
by the organic group.
30 [26] A precipitation mixture comprising the precipitation
promoter of any one of the aforementioned [1] - [25], and an
organic compound protected by an organic group having one or
more aliphatic hydrocarbon groups having not less than 10
carbon atoms.
35 [27] The precipitation mixture of the aforementioned [26],
12
wherein the aliphatic hydrocarbon group having not less than 10
carbon atoms of the organic group is linear.
[28] A method of precipitating an organic compound protected by
an organic group having one or more aliphatic hydrocarbon
groups having not less than 10 carbon atoms 5 in a solvent, by
using the precipitation promoter of any one of the
aforementioned [1] - [25].
[29] The method of the aforementioned [28], wherein the solvent
comprises a polar solvent.
10 [30] The method of the aforementioned [29], wherein the polar
solvent is acetonitrile.
[31] The method of any one of the aforementioned [28] - [30],
wherein the aliphatic hydrocarbon group having not less than 10
carbon atoms of the organic group is linear
15 [32] A production method of an oligonucleotide, comprising a
step of adding a polar solvent to a reaction solution
comprising an oligonucleotide wherein at least one group is
protected by an organic group having one or more aliphatic
hydrocarbon groups having not less than 10 carbon atoms, and
20 other group is optionally further protected by a protecting
group used in nucleic acid synthesis, and the precipitation
promoter of any one of the aforementioned [1] - [25] in a
nonpolar solvent, and separating a precipitate mixture
comprising the oligonucleotide and the precipitation promoter
25 from the reaction solution.
[33] The production method of the aforementioned [32], wherein
the polar solvent is acetonitrile.
[34] The production method of the aforementioned [32] or [33],
wherein the aliphatic hydrocarbon group having not less than 10
30 carbon atoms of the organic group is linear.
[35] The production method of any one of the above-mentioned
[32] - [34], which is performed by a phosphoramidite method.
[36] A production method of an oligonucleotide, which includes
one repeat of production cycle comprising the following steps
35 (1) - (3), or plural repeats thereof by a phosphoramidite
13
method, which comprises the following step (4) in the first
cycle, the following step (5) in each cycle, and the following
step (6) in each cycle except the final cycle:
(1) a step of obtaining a reaction solution comprising a free-
5’-hydroxy-group form by adding an acid to a 5 reaction solution
comprising a nucleoside or oligonucleotide wherein at least one
group selected from an amino group and an imino group of a
nucleic acid base, 2’- and 3’-hydroxy groups of a ribose
residue, and 3’-hydroxy group of a deoxyribose residue is
10 protected by an organic group having one or more aliphatic
hydrocarbon groups having not less than 10 carbon atoms, a 5’-
hydroxy group is protected by a temporary protecting group
removable under acidic conditions, and other group is
optionally further protected by a protecting group used in
15 nucleic acid synthesis in a nonpolar solvent, to deprotect the
temporary protecting group of the 5’-hydroxy group, and
neutralizing same with a base;
(2) a step of obtaining a reaction solution comprising a
phosphite triester form, by adding nucleoside or
20 oligonucleotide wherein a 3’-hydroxy group is phosphoramidited,
a 5’-hydroxy group is protected by a temporary protecting group
removable under acidic conditions, and other group is
optionally further protected by a protecting group used in
nucleic acid synthesis to the reaction solution comprising the
25 free-5’-hydroxy-group form in a nonpolar solvent;
(3) a step of obtaining a reaction solution comprising an
oligonucleotide wherein at least one group selected from an
amino group and an imino group of a nucleic acid base, 2’- and
3’-hydroxy groups of a ribose residue, and 3’-hydroxy group of
30 a deoxyribose residue is protected by an organic group having
one or more aliphatic hydrocarbon groups having not less than
10 carbon atoms, a 5’-hydroxy group is protected by a temporary
protecting group removable under acidic conditions, and other
group is optionally further protected by a protecting group
35 used in nucleic acid synthesis, by adding an oxidizing agent or
14
a sulfurizing agent to the reaction solution comprising the
phosphite triester form in a nonpolar solvent;
(4) a step of adding the precipitation promoter of any one of
the aforementioned [1] - [25] to the reaction solution at any
of before step (1), between steps (1) and 5 (2), between steps
(2) and (3) and after step (3);
(5) a step of separating a precipitation mixture comprising the
free-5’-hydroxy-group form, the phosphite triester form or the
oligonucleotide, and the precipitation promoter from the
10 reaction solution by adding a polar solvent to the reaction
solution comprising the precipitation promoter at after step
(4), and any of between steps (1) and (2), between steps (2)
and (3) and after step (3);
(6) a step of adding a nonpolar solvent to the precipitation
15 mixture obtained in step (5) to give a reaction solution.
[37] The production method of the aforementioned [36], wherein
the polar solvent is acetonitrile.
[38] The production method of the aforementioned [36] or [37],
wherein the aliphatic hydrocarbon group having not less than 10
20 carbon atoms of the organic group is linear.
[39] The method of any one of the above-mentioned [36] - [38],
wherein the steps (5) and (6) are performed after step (3).
[40] The method of any one of the above-mentioned [36] - [39],
further comprising the following step (7):
25 (7) a step of removing all protecting groups of the
oligonucleotide and isolating the oligonucleotide.
[41] The method of any one of the above-mentioned [36] - [40],
wherein the non-polar solvent is a solvent selected from the
group consisting of a halogenated solvent, an aromatic solvent,
30 an ester solvent, an aliphatic solvent, and a combination
thereof.
[Effect of the Invention]
[0023]
Using the precipitation promoter of the present invention,
35 precipitation of an organic compound having a pseudo solid
15
phase protecting group, which is a hydrophobic group, in a
solvent can be promoted, and the recovery rate thereof can be
improved.
[Description of Embodiments]
5 [0024]
[term]
Unless otherwise specified in the sentences, any
technical terms and scientific terms used in the present
specification, have the same meaning as those generally
10 understood by those of ordinary skill in the art the present
invention belongs to. Any methods and materials similar or
equivalent to those described in the present specification can
be used for practicing or testing the present invention, and
preferable methods and materials are described in the following.
15 All publications and patents referred to in the specification
are hereby incorporated by reference so as to describe and
disclose constructed products and methodology described in, for
example, publications usable in relation to the described
invention.
20 [0025]
In the present specification, the “nucleoside” to be the
constituent unit of oligonucleotide means a compound wherein a
nucleic acid base is bonded to the 1’-position of a sugar (e.g.,
ribose or deoxyribose wherein the carbon atoms at 2-position
25 and carbon atom at 4-position of 2-deoxyribose, ribose, ribose
ring or deoxyribose ring are bonded by a divalent organic group,
and the like) by N-glycosidation. Examples of the ribose or
deoxyribose wherein the carbon atom at 2-position and carbon
atom at 4-position of the ribose ring or deoxyribose ring are
30 bonded by a divalent organic group include the following
compounds.
[0026]
16
[0027]
In the present specification, the “nucleotide” means a
compound wherein a phosphoric acid group is bonded to a
5 nucleoside.
In the present specification, the “oligonucleotide” means
a compound wherein one or more nucleotides are bonded to a
nucleoside. While the number of nucleosides in oligonucleotide
in the present invention is not particularly limited, it is
10 preferably 3 - 50, more preferably 5 - 30.
[0028]
In the present specification, the “nucleoside” and
“nucleotide” encompass morpholino nucleoside and morpholino
nucleoside having a morpholine residue instead of a ribose
15 residue or a deoxyribose residue.
In the present specification, the “morpholino nucleoside”
is a compound represented by the following formula (1). In the
present specification, morpholino nucleoside represented by the
formula (1) is referred to as morpholino nucleoside (1).
20 Compounds represented by other formulas may also be referred to
in the same manner.
[0029]
17
[0030]
wherein Base is an optionally protected nucleic acid base.
Morpholino nucleoside (1) can be prepared by a method
known per se (e.g., the method described in 5 WO 91/09033A1), or
a method analogous thereto. Specifically, as shown in the
following scheme, the corresponding ribonucleoside (2) is
subjected to oxidative ring opening with sodium periodate etc.
to give the corresponding 2’,3’-dialdehyde (3), the dialdehyde
10 (3) is subjected to ring closure with ammonia to give 2’,3’-
dihydroxymorpholino nucleoside (4), and dihydroxymorpholino
nucleoside (4) is reduced with a reducing agent (e.g., sodium
cyanoborohydride, sodium triacetoxyborohydride and the like),
whereby morpholino nucleoside (1) can be obtained.
15 [0031]
[0032]
reduction
18
In the present specification, the position number (1’, 2’
and the like) of morpholino nucleoside corresponds to that of
the carbon atom of ribose of ribonucleoside (2) as the material.
In the present specification, according to the usual practice
in the nucleic acid chemistry, morpholino 5 nucleoside on the
terminal of the side having a free 5’-hydroxy group of
morpholino oligonucleotide is referred to as the “5’-terminus”,
and morpholino nucleoside on the terminal of the opposite side
is referred to as the “3’-terminus”.
10 [0033]
In the present specification, the “nucleic acid base” is
not particularly limited as long as it can be used for the
synthesis of nucleic acid and includes, for example, a
pyrimidine base such as cytosyl group, uracil group, thyminyl
15 group and the like, and a purine base such as adenyl group,
guanyl group and the like. The “optionally protected nucleic
acid base” means, for example, that an amino group may be
protected in an adenyl group, a guanyl group or a cytosyl group,
which is a nucleic acid base having an amino group, and a
20 nucleic acid base wherein the amino group therein is protected
by a protecting group sustainable under the deprotection
conditions of the 5’-position of nucleotide or deprotection
conditions of the morpholine ring nitrogen atom of morpholino
nucleotide is preferable. The “amino-protecting group” is not
25 particularly limited, and examples thereof include the
protecting groups described in Greene’s PROTECTIVE GROUPS IN
ORGANIC SYNTHESIS, 4th edition, Wiley-Interscience, 2006 and
the like. Specific examples of the “amino-protecting group”
include a pivaloyl group, a pivaloyloxymethyl group, a
30 trifluoroacetyl group, a phenoxyacetyl group, a 4-
isopropylphenoxyacetyl group, a 4-tert-butylphenoxyacetyl group,
an acetyl group, a benzoyl group, an isobutyryl group, a
dimethylformamidinyl group, a 9-fluorenylmethyloxycarbonyl
group and the like. Among them, a phenoxyacetyl group, a 4-
35 isopropylphenoxyacetyl group, an acetyl group, a benzoyl group,
19
an isobutyryl group and a dimethylformamidinyl group are
preferable. In addition, the carbonyl group of the nucleic
acid base is optionally protected, and can be protected, for
example, by reacting phenol, 2,5-dichlorophenol, 3-chlorophenol,
3,5-dichlorophenol, 2-formylphenol, 2-naphthol, 5 4-methoxyphenol,
4-chlorophenol, 2-nitrophenol, 4-nitrophenol, 4-
acetylaminophenol, pentafluorophenol, 4-pivaloyloxybenzyl
alcohol, 4-nitrophenethyl alcohol, 2-(methylsulfonyl)ethanol,
2-(phenylsulfonyl)ethanol, 2-cyanoethanol, 2-
10 (trimethylsilyl)ethanol, dimethylcarbamoyl chloride,
diethylcarbamoyl chloride, ethylphenylcarbamoyl chloride, 1-
pyrrolidinecarbonyl chloride, 4-morpholinecarbonyl chloride,
diphenylcarbamoyl chloride and the like. In some cases, the
carbonyl-protecting group does not need to be particularly
15 introduced. Moreover, in addition to the above-mentioned
groups, a modified nucleic acid base (e.g., a 8-bromoadenyl
group, a 8-bromoguanyl group, a 5-bromocytosyl group, a 5-
iodocytosyl group, a 5-bromouracil group, a 5-iodouracil group,
a 5-fluorouracil group, a 5-methylcytosyl group, a 8-oxoguanyl
20 group, a hypoxanthinyl group etc.), which is a nucleic acid
base substituted by any 1 to 3 substituents (e.g., a halogen
atom, an alkyl group, an aralkyl group, an alkoxy group, an
acyl group, an alkoxyalkyl group, a hydroxy group, an amino
group, monoalkylamino, dialkylamino, carboxy, cyano, nitro
25 etc.) at any position(s), are also encompassed in the “nucleic
acid base”.
[0034]
In the present specification, the “halogen atom” means a
fluorine atom, a chlorine atom, a bromine atom or iodine atom.
30 [0035]
In the present specification, examples of the “alkyl
(group)” include a linear or branched chain alkyl group having
one or more carbon atoms. When the carbon number is not
particularly limited, it is preferably a C1-10 alkyl group, more
35 preferably a C1-6 alkyl group. When the carbon number is not
20
particularly limited, for example, methyl, ethyl, propyl,
isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl,
hexyl and the like are preferable, and methyl and ethyl are
particularly preferable.
In the present specification, “Ca-b” means 5 that the carbon
number is not less than a and not more than b (a, b each show
an integer).
[0036]
In the present specification, examples of the “aralkyl
10 (group)” include a C7-20 aralkyl group, preferably a C7-16
aralkyl group (a C6-10 aryl-C1-6 alkyl group). Preferable
specific examples include benzyl, 1-phenylethyl, 2-phenylethyl,
1-phenylpropyl, naphthylmethyl, 1-naphthylethyl, 1-
naphthylpropyl and the like, and benzyl is particularly
15 preferable.
[0037]
In the present specification, examples of the “alkoxy
(group)” include an alkoxy group having one or more carbon
atoms. When the carbon number is not particularly limited, it
20 is preferably a C1-10 alkoxy group, more preferably a C1-6 alkoxy
group. When the carbon number is not particularly limited,
methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, secbutoxy,
tert-butoxy, pentyloxy, hexyloxy and the like are
preferable, and methoxy and ethoxy are particularly preferable.
25 [0038]
In the present specification, examples of the “acyl
(group)” include a linear or branched chain C1-6 alkanoyl group,
a C7-13 aroyl group and the like. Specific examples thereof
include formyl, acetyl, n-propionyl, isopropionyl, n-butyryl,
30 isobutyryl, pivaloyl, valeryl, hexanoyl, benzoyl, naphthoyl,
levulinyl and the like, each of which is optionally substituted.
[0039]
In the present specification, as the “alkenyl (group)”, a
linear or branched chain C2-6 alkenyl group and the like are
35 preferable. Preferable examples thereof include vinyl, 1-
21
propenyl, allyl, isopropenyl, butenyl, isobutenyl and the like.
Among them, a C2-4 alkenyl group is preferable.
[0040]
In the present specification, as the “alkynyl (group)”,
include a C2-6 alkynyl group and the like 5 are preferable.
Preferable examples thereof include ethynyl, 1-propynyl, 2-
propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-
pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-
hexynyl, 4-hexynyl, 5-hexynyl and the like. Among them, a C2-4
10 alkynyl group is preferable.
[0041]
In the present specification, the “cycloalkyl (group)”
means a cyclic alkyl group, and examples thereof include
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,
15 cyclooctyl and the like. Among them, a C3-6 cycloalkyl group
such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and
the like is preferable, and cyclohexyl is particularly
preferable.
[0042]
20 In the present specification, the “aryl (group)” means an
aromatic monocyclic or polycyclic (fused) hydrocarbon group.
Specific examples thereof include a C6-14 aryl group such as
phenyl, 1-naphthyl, 2-naphthyl, biphenylyl, 2-anthryl and the
like, and the like. Among them, a C6-10 aryl group is more
25 preferably and phenyl is particularly preferable.
[0043]
In the present specification, examples of the
“hydrocarbon group” include an aliphatic hydrocarbon group, an
aromatic-aliphatic hydrocarbon group, a monocyclic saturated
30 hydrocarbon group, an aromatic hydrocarbon group and the like.
Specific examples thereof include a monovalent group such as an
alkyl group, an alkenyl group, an alkynyl group, a cycloalkyl
group, an aryl group, an aralkyl group and the like, and a
divalent group derived therefrom.
35 [0044]
22
In the present specification, the three alkyl groups in
the “trialkylsilyl (group)” may be the same or different. The
alkyl group may be any of independently linear and branched
chain. As the “trialkylsilyl (group)”, a tri(C1-6 alkyl)silyl
group is preferable, for example, trimethylsilyl 5 group,
triethylsilyl group, triisopropylsilyl group, tertbutyldimethylsilyl
group can be mentioned.
[0045]
In the present specification, the “alkylene (group)” may
10 be any of linear and branched chain. As the “alkylene (group)”,
an alkylene group having one or more carbon atoms can be
mentioned. When the range of carbon number is not particularly
limited, it is preferably a C1-10 alkylene group, more
preferably a C1-6 alkylene group. Preferable specific examples
15 include methylene, ethylene, propylene, butylene, pentylene and
hexylene, particularly preferably methylene and ethylene.
[0046]
In the present specification, examples of the “linker”
include -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH-, -NHC(=O)-,
20 -S-, -SO-, -SO2 -, -Si(R)(R’)O-, -Si(R)(R’)- (R, R’ are each
independently a hydrogen atom or a C1-22 hydrocarbon group) and
the like.
[0047]
In the present specification, the “substituent” of
25 “optionally substituted” encompasses the aforementioned halogen
atom, alkyl group, aralkyl group, alkoxy group, acyl group,
alkenyl group, alkynyl group, cycloalkyl group, aryl group, as
well as hydroxy group, nitro group, cyano group, guanidyl group,
carboxy group, alkoxycarbonyl group (alkoxy moiety is the same
30 as that in the aforementioned alkoxy group), sulfo group,
phospho group, alkylthio group (alkyl moiety is the same as
that in the aforementioned alkyl group), alkylsulfinyl group
(alkyl moiety is the same as that in the aforementioned alkyl
group), alkylsulfonyl group (alkyl moiety is the same as that
35 in the aforementioned alkyl group), amino group, monoalkylamino
23
group (alkyl moiety is the same as that in the aforementioned
alkyl group), dialkylamino group (alkyl moiety is the same as
that in the aforementioned alkyl group), oxo group, acylamino
group (acyl moiety is the same as that in the aforementioned
alkyl group) 5 and the like.
[0048]
In the present specification, examples of the “C3-14
hydrocarbon ring” (including “C3-14 hydrocarbon ring” of the
“optionally substituted C3-14 hydrocarbon ring”) include C3-10
10 cycloalkane, C3-10 cycloalkene, C6-14 aromatic hydrocarbon ring.
Examples of the “C3-10 cycloalkane” include cyclopropane,
cyclobutane, cyclopentane, cyclohexane, cycloheptane,
cyclooctane.
Examples of the “C3-10 cycloalkene” include cyclopropene,
15 cyclobutene, cyclopentene, cyclohexene, cycloheptene,
cyclooctene.
Examples of the “C6-14 aromatic hydrocarbon ring” include
benzene, naphthalene.
[0049]
20 In the present specification, examples of the “monocyclic
heterocyclic group” (including “monocyclic heterocyclic group”
of the “optionally substituted monocyclic heterocyclic group”)
include monocyclic aromatic heterocyclic group and monocyclic
nonaromatic heterocyclic group each containing, as a ring25
constituting atom besides carbon atom, 1 – 4 hetero atoms
selected from nitrogen atom, sulfur atom and oxygen atom.
Examples of the “monocyclic aromatic heterocyclic group”
include 5- or 6-membered monocyclic aromatic heterocyclic
groups such as thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl,
30 thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl,
pyrazinyl, pyrimidinyl, pyridazinyl, 1,2,4-oxadiazolyl, 1,3,4-
oxadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, triazolyl,
tetrazolyl, triazinyl and the like.
Examples of the “monocyclic nonaromatic heterocyclic
35 group” include 3- to 8-membered monocyclic nonaromatic
24
heterocyclic groups such as aziridinyl, oxiranyl, thiiranyl,
azetidinyl, oxetanyl, thietanyl, tetrahydrothienyl,
tetrahydrofuranyl, pyrrolinyl, pyrrolidinyl, imidazolinyl,
imidazolidinyl, oxazolinyl, oxazolidinyl, pyrazolinyl,
pyrazolidinyl, thiazolinyl, 5 thiazolidinyl,
tetrahydroisothiazolyl, tetrahydrooxazolyl,
tetrahydroisooxazolyl, piperidinyl, piperazinyl,
tetrahydropyridinyl, dihydropyridinyl, dihydrothiopyranyl,
tetrahydropyrimidinyl, tetrahydropyridazinyl, dihydropyranyl,
10 tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl (e.g.,
morpholin-4-yl), thiomorpholinyl, azepanyl, diazepanyl,
azepinyl, oxepanyl, azocanyl, diazocanyl and the like.
[0050]
[precipitation promoter]
15 The precipitation promoter of the present invention is an
organic compound having one or more linear aliphatic
hydrocarbon groups having not less than 10 carbon atoms wherein
the aforementioned aliphatic hydrocarbon group has not less
than 20 carbon atoms in total, which is used to precipitate an
20 organic compound protected by an organic group having one or
more aliphatic hydrocarbon groups having not less than 10
carbon atoms (hereinafter sometimes referred to as “the pseudo
solid phase protecting group of the present invention”) in a
solvent. Only one kind of the precipitation promoter of the
25 present invention may be used, or two or more kinds thereof may
be used in combination.
[0051]
The aforementioned solvent is preferably a solvent
containing a polar solvent, more preferably a mixed solvent of
30 a polar solvent and a nonpolar solvent. The aforementioned
polar solvent is preferably acetonitrile.
[0052]
The linear aliphatic hydrocarbon group having not less
than 10 carbon atoms contained in the precipitation promoter of
35 the present invention needs to have not less than 20,
25
preferably 22 or more, carbon atoms in total. When the total
carbon number is less than 20, the organic compound protected
by a pseudo solid phase protecting group in the liquid phase
cannot be precipitated sufficiently, and the recovery rate of
the organic compound decreases. While the 5 upper limit is not
particularly limited, the total carbon number is preferably not
more than 220, more preferably not more than 100, further
preferably not more than 70.
[0053]
10 The linear aliphatic hydrocarbon group having not less
than 10 carbon atoms in the precipitation promoter of the
present invention is preferably a group selected from a linear
C10-40 alkyl group and a linear C10-40 alkenyl group, more
preferably a linear C10-40 alkyl group, further preferably a
15 linear C10-30 alkyl group, particularly preferably a linear C12-28
alkyl group, most preferably a linear C14-26 alkyl group. The
number of a linear aliphatic hydrocarbon group having not less
than 10 carbon atoms contained in the precipitation promoter of
the present invention is preferably 1 - 12, more preferably 1 -
20 9.
[0054]
One embodiment of the precipitation promoter of the
present invention is an organic compound having one or more
structures represented by the following formula (G):
25 [0055]
[0056]
wherein
each R1 is independently a linear C10-40 alkyl group;
each X1 30 is independently a single bond, -O-, -C(=O)-, -
C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
ring A1 is an optionally substituted C3-14 hydrocarbon
26
ring; and
n is an integer of 1 - 4.
[0057]
In the formula (G), the bonding direction of X1
corresponds to the direction of the above-5 mentioned chemical
formula. For example, when X1 is -C(=O)O-, it is bonded to R1
on the left side of -C(=O)O-, and bonded to A1 on the right
side of -C(=O)O-.
[0058]
When the formula (G) contains a plurality of R110 , they may
be the same or different. The same applied to X1.
[0059]
A preferable one embodiment of the precipitation promoter
of the present invention is a compound represented by the
15 following formula (I). The “compound represented by the
formula (I)” is sometimes abbreviated as “compound (I)”.
Compounds represented by other formulas may also be abbreviated
similarly.
[0060]
20
[0061]
wherein
each R1 is independently a linear C10-40 alkyl group;
each X1 is independently a single bond, -O-, -C(=O)-, -
25 C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
ring A1 and ring B1 are each independently an optionally
substituted C3-14 hydrocarbon ring;
each X2 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q-, -(CH2)p-OC(=O)-
30 (CH2)q-, -(CH2)p-C(=O)NH-(CH2)q- or -(CH2)p-NHC(=O)-(CH2)q- (p and
q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-, -(CH2)r-
C(=O)O-, -(CH2)r-OC(=O)-, -(CH2)r-C(=O)NH- or -(CH2)r-NHC(=O)- (r
27
is an integer of 0 - 3);
R2 is a hydrogen atom, an optionally substituted C3-8
cycloalkyl group, an optionally substituted C1-6 alkyl group, an
optionally substituted C2-6 alkenyl group, an optionally
substituted C2-6 alkynyl group, an optionally 5 substituted C6-14
aryl group, an optionally substituted monocyclic heterocyclic
group or a tri(C1-6 alkyl)silyl group (preferably hydrogen atom,
an optionally substituted C3-8 cycloalkyl group, an optionally
substituted C1-6 alkyl group, an optionally substituted C2-6
10 alkenyl group, an optionally substituted C2-6 alkynyl group, an
optionally substituted C6-14 aryl group or an optionally
substituted monocyclic heterocyclic group);
n and m are each independently an integer of 1 - 4;
m’ is 0 or 1 when m is 1, and 1 when m is 2, 3 or 4.
15 [0062]
In the formula (I), the bonding directions of X1, X2 and
X3 each correspond to the direction of the above-mentioned
chemical formula. For example, when X3 is -(CH2)r-O-, it is
bonded to ring B1 on the left side of -(CH2)r-O-, and bonded to
R2 20 on the right side of -(CH2)r-O-.
[0063]
When the formula (I) contains a plurality of R1, they may
be the same or different. The same applied to X1, ring A1, and
X2.
25 [0064]
A more preferable one embodiment of the precipitation
promoter of the present invention is a compound represented by
the following formula (II).
[0065]
30
[0066]
28
wherein
each R1 is independently a linear C10-40 alkyl group;
each X1 is independently a single bond, -O-, -C(=O)-, -
C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
ring A2 and ring B1 are each independently 5 an optionally
substituted C3-14 hydrocarbon ring;
each X2 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q-, -(CH2)p-OC(=O)-
(CH2)q-, -(CH2)p-C(=O)NH-(CH2)q- or -(CH2)p-NHC(=O)-(CH2)q- (p and
10 q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-, -(CH2)r-
C(=O)O-, -(CH2)r-OC(=O)-, -(CH2)r-C(=O)NH- or -(CH2)r-NHC(=O)- (r
is an integer of 0 - 3);
R2 is a hydrogen atom, an optionally substituted C3-8
15 cycloalkyl group, an optionally substituted C1-6 alkyl group, an
optionally substituted C2-6 alkenyl group, an optionally
substituted C2-6 alkynyl group, an optionally substituted C6-14
aryl group, an optionally substituted monocyclic heterocyclic
group or a tri(C1-6 alkyl)silyl group (preferably hydrogen atom,
20 an optionally substituted C3-8 cycloalkyl group, an optionally
substituted C1-6 alkyl group, an optionally substituted C2-6
alkenyl group, an optionally substituted C2-6 alkynyl group, an
optionally substituted C6-14 aryl group or an optionally
substituted monocyclic heterocyclic group);
25 m is an integer of 1 to 4; and
m’ is 0 or 1 when m is 1, and 1 when m is 2, 3 or 4.
[0067]
In the formula (II), the bonding directions of X1, X2 and
X3 each correspond to the direction of the above-mentioned
chemical formula. For example, when X3 30 is -(CH2)r-O-, it is
bonded to ring B1 on the left side of -(CH2)r-O-, and bonded to
R2 on the right side of -(CH2)r-O-.
[0068]
When the formula (II) contains a plurality of R1, they
may be the same or different. The same applied to X135 . When
29
the formula (II) contains a plurality of ring A2, they may be
the same or different. The same applied to X2.
[0069]
When R1 is present in plurality, they may be the same or
different. Each R1 is independently preferably 5 a linear C10-30
alkyl group, more preferably a linear C12-28 alkyl group,
further preferably a linear C14-26 alkyl group.
[0070]
In one embodiment of the present invention, R2 is
10 preferably a hydrogen atom, an optionally substituted C3-8
cycloalkyl group, an optionally substituted C1-6 alkyl group, an
optionally substituted C2-6 alkenyl group, an optionally
substituted C2-6 alkynyl group, an optionally substituted C6-14
aryl group or an optionally substituted monocyclic heterocyclic
15 group, more preferably a hydrogen atom, an optionally
substituted C1-6 alkyl group, an optionally substituted C2-6
alkenyl group, an optionally substituted C2-6 alkynyl group or
an optionally substituted C6-14 aryl group, further preferably a
hydrogen atom or an optionally substituted C1-6 alkyl group,
20 particularly preferably a hydrogen atom or a C1-6 alkyl group.
[0071]
In another embodiment of the present invention, R2 is
preferably a hydrogen atom, an optionally substituted C1-6 alkyl
group, an optionally substituted C2-6 alkenyl group, an
25 optionally substituted C2-6 alkynyl group or an optionally
substituted C6-14 aryl group, an optionally substituted 3 - 8-
membered monocyclic nonaromatic heterocyclic group or a tri(C1-6
alkyl)silyl group, more preferably hydrogen atom or an
optionally substituted C1-6 alkyl group or a 3- to 8-membered
30 monocyclic nonaromatic heterocyclic group or a tri(C1-6
alkyl)silyl group, further preferably a hydrogen atom, a C1-6
alkyl group optionally substituted by a C1-6 alkanoylamino group
(e.g., acetylamino group) or a 3- to 8-membered monocyclic
nonaromatic heterocyclic group (e.g., morpholin-4-yl group) or
35 a tri(C1-6 alkyl)silyl group (e.g., triisopropylsilyl group).
30
[0072]
Each X1 is independently preferably a single bond, -O-, -
C(=O)-, -C(=O)O- or -OC(=O)-, more preferably -O-, -C(=O)O- or
-OC(=O)-, further preferably -O-.
5 [0073]
Each X2 is independently preferably -(CH2)p-, -(CH2)p-O-
(CH2)q-, -(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q- or -(CH2)p-
OC(=O)-(CH2)q- (p and q are each independently an integer of 0
- 3), more preferably -(CH2)p-, -(CH2)p-O-(CH2)q-, -(CH2)p-C(=O)-
10 (CH2)q-, -(CH2)p-C(=O)O-(CH2)q- or -(CH2)p-OC(=O)-(CH2)q- (p and q
are each independently an integer of 0 - 3), further preferably
a single bond, -(CH2)p-O-(CH2)q-, -C(=O)-, -C(=O)O-(CH2)q- or -
(CH2)p-OC(=O)- (p and q are each independently an integer of 0
or 1).
15 [0074]
X3 is preferably a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-,
-(CH2)r-C(=O)O- or -(CH2)r-OC(=O)- (r is an integer of 0 - 3),
more preferably a single bond, -(CH2)r-C(=O)-, -(CH2)r-C(=O)Oor
-(CH2)r-OC(=O)- (r is an integer of 0 - 3), further
20 preferably a single bond, -C(=O)-, -C(=O)O- or -OC(=O)-.
[0075]
n is preferably an integer of 1 - 3.
m is preferably an integer of 1 - 3, more preferably 1 or
3.
25 [0076]
The C3-14 hydrocarbon ring is preferably selected from a
benzene ring and a cyclohexane ring, more preferably a benzene
ring.
[0077]
30 A preferable compound (I) is a compound wherein
each R1 is independently a linear C10-30 alkyl group;
each X1 is independently a single bond, -O-, -C(=O)-, -
C(=O)O- or -OC(=O)-;
ring A2 and ring B1 are each independently a benzene ring
35 or a cyclohexane ring;
31
each X2 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q- or -(CH2)p-OC(=O)-
(CH2)q- (p and q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-, -(CH2)r-
C(=O)O- or -(CH2)r-OC(=O)- (r is an 5 integer of 0 - 3);
R2 is a hydrogen atom, an optionally substituted C1-6
alkyl group, an optionally substituted C2-6 alkenyl group, an
optionally substituted C2-6 alkynyl group, an optionally
substituted C6-14 aryl group, an optionally substituted 3 - 8-
10 membered monocyclic nonaromatic heterocyclic group or a tri(C1-6
alkyl)silyl group;
n is an integer of 1 - 3;
m is an integer of 1 - 3; and
m’ is 0 or 1 when m is 1, and 1 when m is 2 or 3.
In this embodiment, R2 15 is more preferably a hydrogen atom, an
optionally substituted C1-6 alkyl group, an optionally
substituted C2-6 alkenyl group, an optionally substituted C2-6
alkynyl group or an optionally substituted C6-14 aryl group.
[0078]
20 A more preferable compound (I) is a compound wherein
each R1 is independently a linear C12-28 alkyl group;
each X1 is independently -O-, -C(=O)O- or -OC(=O)-;
ring A2 and ring B1 are each independently a benzene ring
or a cyclohexane ring;
each X2 25 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q- or -(CH2)p-OC(=O)-
(CH2)q- (p and q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-C(=O)-, -(CH2)r-C(=O)O- or -
(CH2)r-OC(=O)- (r is an integer of 0 - 3);
R2 30 is a hydrogen atom, an optionally substituted C1-6
alkyl group or a 3- to 8-membered monocyclic nonaromatic
heterocyclic group or a tri(C1-6 alkyl)silyl group;
n is an integer of 1 - 3;
m is an integer of 1 - 3; and
35 m’ is 0 or 1 when m is 1, and 1 when m is 2 or 3.
32
In this embodiment, it is more preferable that R2 is a hydrogen
atom or an optionally substituted C1-6 alkyl group.
[0079]
A still more preferable compound (I) is a compound
5 wherein
each R1 is independently a linear C12-28 alkyl group;
each X1 is independently -O-, -C(=O)O- or -OC(=O)-;
ring A2 and ring B1 are each independently a benzene ring
or a cyclohexane ring;
each X2 10 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q- or -(CH2)p-OC(=O)-
(CH2)q- (p and q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-C(=O)-, -(CH2)r-C(=O)O- or -
(CH2)r-OC(=O)- (r is an integer of 0 - 3);
R2 15 is a hydrogen atom, a C1-6 alkyl group optionally
substituted by a C1-6 alkanoylamino group (e.g., acetylamino
group) or a 3- to 8-membered monocyclic nonaromatic
heterocyclic group (e.g., morpholin-4-yl group) or a tri(C1-6
alkyl)silyl group (e.g., triisopropylsilyl group);
20 n is an integer of 1 - 3;
m is an integer of 1 - 3; and
m’ is 0 or 1 when m is 1, and 1 when m is 2 or 3.
[0080]
A further preferable compound (I) is a compound wherein
each R1 25 is independently a linear C14-26 alkyl group;
X1 is -O-;
ring A2 and ring B1 are each independently a benzene ring
or a cyclohexane ring;
each X2 is independently a single bond, -(CH2)p-O-(CH2)q-,
30 -C(=O)-, -C(=O)O-(CH2)q- or -(CH2)p-OC(=O)- (p and q are each
independently an integer of 0 or 1);
X3 is a single bond, -C(=O)-, -C(=O)O- or -OC(=O)-;
R2 is a hydrogen atom, a C1-6 alkyl group optionally
substituted by a C1-6 alkanoylamino group (e.g., acetylamino
35 group) or a 3- to 8-membered monocyclic nonaromatic
33
heterocyclic group (e.g., morpholin-4-yl group) or a tri(C1-6
alkyl)silyl group (e.g., triisopropylsilyl group);
n is an integer of 1 - 3;
m is 1 or 3; and
m’ is 0 or 1 when m is 1, and 5 1 when m is 3.
[0081]
In the aforementioned further preferable compound (I), R2
is more preferably a hydrogen atom or a C1-6 alkyl group.
[0082]
10 In the aforementioned further preferable compound (I), it
is more preferable that when m is 1 and m’ is 0, each X2 is
independently a single bond, -C(=O)O-(CH2)q- or -(CH2)p-OC(=O)-
(p and q are each independently an integer of 0 or 1), R2 is a
C1-6 alkyl group.
15 [0083]
In the aforementioned further preferable compound (I), it
is more preferable that when m is 1 or 3 and m’ is 1, each X2
is independently -(CH2)p-O-(CH2)q- or -C(=O)- (p and q are each
independently an integer of 0 or 1), X3 is a single bond or -
C(=O)O-, and R2 20 is a hydrogen atom or a C1-6 alkyl group.
[0084]
A particularly preferable compound (I) is a compound
wherein
each R1 is independently a linear C14-26 alkyl group;
X1 25 is -O-;
ring A2 and ring B1 are each a benzene ring;
each X2 is independently a single bond, -(CH2)p-O-(CH2)q-,
-C(=O)-, -C(=O)O-(CH2)q- or -(CH2)p-OC(=O)- (p and q are each
independently an integer of 0 or 1);
X3 30 is a single bond, -C(=O)-, -C(=O)O- or -OC(=O)-;
R2 is a hydrogen atom or a C1-6 alkyl group;
n is an integer of 1 - 3;
m is 1 or 3; and
m’ is 0 or 1 when m is 1, and 1 when m is 3.
35 [0085]
34
In the aforementioned particularly preferable compound
(I), when m is 1 and m’ is 0, it is more preferable that X2 is
a single bond, -C(=O)O-(CH2)q- or -(CH2)p-OC(=O)- (p and q are
each independently an integer of 0 or 1), and R2 is a C1-6 alkyl
5 group.
[0086]
In the aforementioned particularly preferable compound
(I), when m is 1 or 3 and m’ is 1, it is more preferable that
X2 is -(CH2)p-O-(CH2)q- or -C(=O)- (p and q are each
independently an integer of 0 or 1), and X3 10 is a single bond or
-C(=O)O-.
[0087]
A preferable compound (II) is a compound wherein
each R1 is independently a linear C10-30 alkyl group;
each X1 15 is independently a single bond, -O-, -C(=O)-, -
C(=O)O- or -OC(=O)-;
ring A2 and ring B1 are each independently a benzene ring
or a cyclohexane ring;
each X2 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
20 (CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q- or -(CH2)p-OC(=O)-
(CH2)q- (p and q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-, -(CH2)r-
C(=O)O- or -(CH2)r-OC(=O)- (r is an integer of 0 - 3);
R2 is a hydrogen atom, an optionally substituted C1-6
25 alkyl group, an optionally substituted C2-6 alkenyl group, an
optionally substituted C2-6 alkynyl group, an optionally
substituted C6-14 aryl group, an optionally substituted 3 - 8-
membered monocyclic nonaromatic heterocyclic group or a tri(C1-6
alkyl)silyl group;
30 m is an integer of 1 - 3; and
m’ is 0 or 1 when m is 1, and 1 when m is 2 or 3.
In this embodiment, it is more preferable that R2 is a hydrogen
atom, an optionally substituted C1-6 alkyl group, an optionally
substituted C2-6 alkenyl group, an optionally substituted C2-6
35 alkynyl group or an optionally substituted C6-14 aryl group.
35
[0088]
A more preferable compound (II) is a compound wherein
each R1 is independently a linear C12-28 alkyl group;
each X1 is independently -O-, -C(=O)O- or -OC(=O)-;
ring A2 and ring B1 are each independently 5 a benzene ring
or a cyclohexane ring;
each X2 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q- or -(CH2)p-OC(=O)-
(CH2)q- (p and q are each independently an integer of 0 - 3);
X3 10 is a single bond, -(CH2)r-C(=O)-, -(CH2)r-C(=O)O- or -
(CH2)r-OC(=O)- (r is an integer of 0 - 3);
R2 is a hydrogen atom, an optionally substituted C1-6
alkyl group or a 3- to 8-membered monocyclic nonaromatic
heterocyclic group or a tri(C1-6 alkyl)silyl group;
15 m is an integer of 1 - 3; and
m’ is 0 or 1 when m is 1, and 1 when m is 2 or 3.
In this embodiment, it is more preferable that R2 is a hydrogen
atom or an optionally substituted C1-6 alkyl group.
[0089]
20 A still more preferable compound (II) is a compound
wherein
each R1 is independently a linear C12-28 alkyl group;
each X1 is independently -O-, -C(=O)O- or -OC(=O)-;
ring A2 and ring B1 are each independently a benzene ring
25 or a cyclohexane ring;
each X2 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q- or -(CH2)p-OC(=O)-
(CH2)q- (p and q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-C(=O)-, -(CH2)r-C(=O)O- or -
30 (CH2)r-OC(=O)- (r is an integer of 0 - 3);
R2 is a hydrogen atom, a C1-6 alkyl group optionally
substituted by a C1-6 alkanoylamino group (e.g., acetylamino
group) or a 3- to 8-membered monocyclic nonaromatic
heterocyclic group (e.g., morpholin-4-yl group) or a tri(C1-6
35 alkyl)silyl group (e.g., triisopropylsilyl group);
36
m is an integer of 1 - 3; and
m’ is 0 or 1 when m is 1, and 1 when m is 2 or 3.
[0090]
A further preferable compound (II) is a compound wherein
each R1 is independently a linear 5 C14-26 alkyl group;
X1 is -O-;
ring A2 and ring B1 are each a benzene ring or a
cyclohexane ring;
each X2 is independently a single bond, -(CH2)p-O-(CH2)q-,
10 -C(=O)-, -C(=O)O-(CH2)q- or -(CH2)p-OC(=O)- (p and q are each
independently an integer of 0 or 1);
X3 is a single bond, -C(=O)-, -C(=O)O- or -OC(=O)-;
R2 is a hydrogen atom, a C1-6 alkyl group optionally
substituted by a C1-6 alkanoylamino group (e.g., acetylamino
15 group) or a 3 - 8-membered monocyclic nonaromatic heterocyclic
group (e.g., morpholin-4-yl group) or a tri(C1-6 alkyl)silyl
group (e.g., triisopropylsilyl group);
m is 1 or 3; and
m’ is 0 or 1 when m is 1, and 1 when m is 3.
20 [0091]
In the aforementioned further preferable compound (II),
it is more preferable that R2 is a hydrogen atom or a C1-6 alkyl
group.
[0092]
25 In the aforementioned further preferable compound (II),
it is more preferable that when m is 1 and m’ is 0, each X2 is
independently a single bond, -C(=O)O-(CH2)q- or -(CH2)p-OC(=O)-
(p and q are each independently an integer of 0 or 1), and R2
is a C1-6 alkyl group.
30 [0093]
In the aforementioned further preferable compound (II),
it is more preferable that when m is 1 or 3 and m’ is 1, each
X2 is independently -(CH2)p-O-(CH2)q- or -C(=O)- (p and q are
each independently an integer of 0 or 1), X3 is a single bond
or -C(=O)O-, and R2 35 is a hydrogen atom or a C1-6 alkyl group.
37
[0094]
A particularly preferable compound (II) is a compound
wherein
each R1 is independently a linear C14-26 alkyl group;
X1 5 is -O-;
ring A2 and ring B1 are each a benzene ring;
each X2 is independently a single bond, -(CH2)p-O-(CH2)q-,
-C(=O)-, -C(=O)O-(CH2)q- or -(CH2)p-OC(=O)- (p and q are each
independently an integer of 0 or 1);
X3 10 is a single bond, -C(=O)-, -C(=O)O- or -OC(=O)-;
R2 is a hydrogen atom or a C1-6 alkyl group;
m is 1 or 3; and
m’ is 0 or 1 when m is 1, and 1 when m is 3.
[0095]
15 In the aforementioned particularly preferable compound
(II), it is more preferable that when m is 1 and m’ is 0, X2 is
a single bond, -C(=O)O-(CH2)q- or -(CH2)p-OC(=O)- (p and q are
each independently an integer of 0 or 1), and R2 is a C1-6 alkyl
group.
20 [0096]
In the aforementioned particularly preferable compound
(II), it is more preferable that when m is 1 or 3 and m’ is 1,
X2 is -(CH2)p-O-(CH2)q- or -C(=O)- (p and q are each
independently an integer of 0 or 1), and X3 is a single bond or
25 -C(=O)O-.
[0097]
Specific examples encompassed in the aforementioned
compound (I) include compounds represented by the following
formulas (IV-a) - (IV-n). Among the specific examples,
30 compound (IV-a) - compound (IV-l) are preferable, compound (IVa)
- compound (IV-j) are more preferable, compound (IV-a) -
compound (IV-g) are further preferable, compound (IV-a) -
compound (IV-e) are particularly preferable, and compound (IVa)
and compound (IV-b) are most preferable.
35 [0098]
38
[0099]
[0100]
5
[0101]
[0102]
39
[0103]
[0104]
Another embodiment of the precipitation 5 promoter of the
present invention is optionally substituted C1-10 alkane having
one or more linear C10-40 alkyl groups via a group selected from
the group consisting of -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -
C(=O)NH- and -NHC(=O)- (hereinafter sometimes to be referred to
10 as “alkane compound”).
[0105]
C1-10 alkane may be linear or branched chain. The carbon
number of C1-10 alkane is preferably 1 - 8, more preferably 2 -
6, further preferably 3 - 6.
15 The linear C10-40 alkyl group of the alkane compound is
preferably a linear C10-30 alkyl group, more preferably a linear
C12-28 alkyl group, further preferably a linear C14-26 alkyl group.
The number of the linear C10-40 alkyl groups of the alkane
compound is preferably 2 - 6, more preferably 2 - 4, further
20 preferably 3 or 4.
The group bonding the linear C10-40 alkyl group and C1-10
alkane is preferably a group selected from the group consisting
of -O-, -C(=O)-, -C(=O)O- and -OC(=O)-, more preferably -O-.
[0106]
40
A preferable alkane compound is optionally substituted
C1-8 alkane having 2 – 6 linear C10-30 alkyl groups via a group
selected from the group consisting of -O-, -C(=O)-, -C(=O)Oand
-OC(=O)-. A more preferable alkane compound is optionally
substituted C2-6 alkane having 2 – 4 linear 5 C10-30 alkyl groups
via -O-. A further preferable alkane compound is optionally
substituted C3-6 alkane having 3 or 4 linear C14-26 alkyl groups
via -O-.
[0107]
10 Specific examples of the alkane compound include
compounds represented by the following formulas (V-a) and (Vb):
[0108]
15 [0109]
[Production method of precipitation promoter of the present
invention]
For example, compound (I) wherein X1 is -O-, m is 1, and
m’ is 0 can be produced by alkylation of a hydroxy group of
20 compound (IA-2) by using compound (IA-1) having a halogen atom
(Xa), as in the following formula. Alkylation is well known in
the field of organic synthesis, and those of ordinary skill in
the art can perform alkylation by appropriately setting the
conditions therefor.
25 [0110]
41
[0111]
wherein Xa is a halogen atom (preferably chlorine atom or
bromine atom), and other symbols are as defined above.
For example, compound (I) wherein X2 is -CH2-O-, and m’
is 1 can be produced by alkylation of 5 a hydroxy group of
compound (IB) by using compound (IA-3) having a halogen atom
(Xa), as in the following formula.
[0112]
10 [0113]
wherein each symbol is as defined above.
Other precipitation promoter of the present invention can
also be produced according to a method known per se (e.g.,
alkylation, esterification, amidation etc.) or a method
15 analogous thereto from a starting compound. As the starting
compound, a commercially available product may be used, or a
compound produced according to a method known per se or a
method analogous thereto may be used.
[0114]
20 [Pseudo solid phase protecting group of the present invention]
The precipitation promoter of the present invention is
used to precipitate an organic compound protected by an organic
group having one or more aliphatic hydrocarbon groups having
not less than 10 carbon atoms (namely, the pseudo solid phase
25 protecting group of the present invention) from a solvent
containing the compound. Specific examples of the pseudo solid
phase protecting group of the present invention and an organic
compound protected by the pseudo solid phase protecting group
42
include nucleoside, nucleotide and the like disclosed in WO
2012/157723, WO 2012/157723, WO 2014/189142, JP-A-2010-116418
and the like, and amino acid, peptide and the like disclosed in
WO 2010/104169, WO 2010/113939, WO 2011/078295, WO 2012/029794,
JP-A-2009-185063, JP-A-2010-275254 and the 5 like. The pseudo
solid phase protecting group of the present invention is
preferably an organic group not removed under acidic conditions
but removed under basic conditions.
[0115]
10 One embodiment of the pseudo solid phase protecting group
of the present invention is, for example, an organic group
having a C6-14 aromatic hydrocarbon ring, which is bonded via a
linker to a hydrocarbon group bonded via a single bond or a
linker to an aliphatic hydrocarbon group having not less than
15 10 carbon atoms. As the aforementioned hydrocarbon group,
those mentioned above can be recited, and an aliphatic
hydrocarbon group, an araliphatic hydrocarbon group (e.g.,
benzyl group) and a group (e.g., cyclohexylmethyl group)
wherein a monocyclic saturated hydrocarbon group is bonded to
20 an aliphatic hydrocarbon group are preferable. The
“hydrocarbon group bonded via a single bond or a linker to an
aliphatic hydrocarbon group having not less than 10 carbon
atoms” also includes “an aliphatic hydrocarbon group having not
less than 10 carbon atoms” itself (i.e., “aliphatic hydrocarbon
25 group bonded via a single bond to aliphatic hydrocarbon group”).
[0116]
While the aliphatic hydrocarbon group having not less
than 10 carbon atoms in the pseudo solid phase protecting group
of the present invention may be linear or branched chain, it is
30 preferably linear to sufficiently exhibit the effect of the
precipitation promoter of the present invention. The aliphatic
hydrocarbon group having not less than 10 carbon atoms is more
preferably a group selected from a linear C10-40 alkyl group and
a linear C10-40 alkenyl group, still more preferably a linear
35 C10-40 alkyl group, further preferably a linear C10-30 alkyl group,
43
particularly preferably a linear C12-28 alkyl group, and most
preferably a linear C14-26 alkyl group.
[0117]
The aforementioned linker is preferably selected from -O-,
-C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH-, 5 -NHC(=O)-, -S-, -SO-, -
SO2 -, -Si(R)(R’)O-, -Si(R)(R’)- (R, R’ are each independently
a hydrogen atom or a C1-22 hydrocarbon group), more preferably
selected from -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH- and -
NHC(=O)-, and more preferably -O-.
10 [0118]
The aforementioned C6-14 aromatic hydrocarbon ring is
preferably selected from a benzene ring and a naphthalene ring,
and more preferably a benzene ring.
[0119]
15 The aforementioned “C6-14 aromatic hydrocarbon ring, which
is bonded via a linker to a hydrocarbon group bonded via a
single bond or a linker to an aliphatic hydrocarbon group
having not less than 10 carbon atoms” is preferably a “C6-14
aromatic hydrocarbon ring, which is bonded via a linker to a
20 linear aliphatic hydrocarbon group having not less than 10
carbon atoms”, more preferably a “C6-14 aromatic hydrocarbon
ring, which is bonded via a linker to a group selected from a
linear C10-40 alkyl group and a linear C10-40 alkenyl group”,
still more preferably a “C6-14 aromatic hydrocarbon ring, which
25 is bonded via a linker to a linear C10-40 alkyl group”, further
preferably a “C6-14 aromatic hydrocarbon ring, which is bonded
via a linker to a linear C10-30 alkyl group”, particularly
preferably a “C6-14 aromatic hydrocarbon ring, which is bonded
via a linker to a linear C12-28 alkyl group”, and most
30 preferably a “C6-14 aromatic hydrocarbon ring, which is bonded
via a linker to a linear C14-26 alkyl group”. In these groups,
the linker is preferably -O-. In these groups, the C6-14
aromatic hydrocarbon ring is preferably a benzene ring.
[0120]
35 The aforementioned pseudo solid phase protecting group is
44
preferably an “organic group having a benzene ring, which is
bonded via -O- to a hydrocarbon group bonded via a single bond
or -O- to a linear C10-40 alkyl group”, more preferably an
“organic group having a benzene ring, which is bonded via -Oto
a linear C10-40 alkyl group”, further preferably 5 an “organic
group having a benzene ring, which is bonded via -O- to a
linear C10-30 alkyl group”, particularly preferably an “organic
group having a benzene ring, which is bonded via -O- to a
linear C12-28 alkyl group”, most preferably an “organic group
10 having a benzene ring, which is bonded via -O- to a linear C14-
26 alkyl group”.
[0121]
The pseudo solid phase protecting group of the present
invention is, for example, a group represented by the following
15 formula (III) (hereinafter to be referred to as “pseudo solid
phase protecting group (III)”):
[0122]
20 [0123]
wherein
** shows a bonding position to a group to be protected;
L is a single bond, or a group represented by the formula (a1)
or (a1’):
25 [0124]
[0125]
wherein
* shows the bonding position to Y;
** 30 is as defined above;
45
R8 and R9 are each independently a C1-22 hydrocarbon group;
L1 is a divalent C1-22 hydrocarbon group; and
L2 is a single bond, or **C(=O)N(R2’)-R1’-N(R3)*** wherein ** shows
the bonding position to L1, *** shows the bonding position to
C=O, R1’ is a C1-22 alkylene group, R2’ and R3 5 are each
independently a hydrogen atom or a C1-22 alkyl group, or R2’ and
R3 are optionally joined to form a ring,
Y is a single bond, an oxygen atom, or NR wherein R is a
hydrogen atom, an alkyl group or an aralkyl group, and
10 Z is a group represented by the formula (a2), formula
(a2’) or formula (a2”) {preferably formula (a2) or formula
(a2’)}:
[0126]
15 [0127]
wherein
* shows a bonding position;
R4 is a hydrogen atom, or when Rb is a group represented
by the following formula (a3), optionally joined with R6 of
20 ring C to show a single bond or -O- and to form a fused ring
(preferably fluorene ring or xanthene ring) together with ring
A or ring B and ring C;
Q in the number of k are each independently -O-, -C(=O)-,
-C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
R5 25 in the number of k are each independently a
hydrocarbon group bonded via a single bond or a linker to an
aliphatic hydrocarbon group having not less than 10 carbon
atoms (preferably a hydrocarbon group bonded via a single bond
or a linker to a linear aliphatic hydrocarbon group having not
30 less than 10 carbon atoms);
46
k is an integer of 1 to 4;
ring A and ring B each independently optionally further
have, in addition to QR5 in the number of k, a substituent
selected from the group consisting of a halogen atom, a C1-6
alkyl group optionally substituted by a halogen 5 atom(s), and a
C1-6 alkoxy group optionally substituted by a halogen atom(s);
Ra is a hydrogen atom, or a phenyl group optionally
substituted by a halogen atom (preferably, a hydrogen atom);
and
10 Rb is a hydrogen atom, or a group represented by the
formula (a3):
[0128]
[0129]
wherein * 15 shows a bonding position;
j is an integer of 0 to 4;
Q in the number of j are each independently as defined
above;
R7 in the number of j are each independently a
20 hydrocarbon group bonded via a single bond or a linker to an
aliphatic hydrocarbon group having not less than 10 carbon
atoms (preferably, a hydrocarbon group bonded via a single bond
or a linker to a linear aliphatic hydrocarbon group having not
less than 10 carbon atoms);
R6 is a hydrogen atom, or optionally joined with R4 25 of
ring A or ring B to show a single bond or -O- and to form a
fused ring (preferably fluorene ring or xanthene ring) together
with ring A or ring B and ring C; and
ring C optionally further has, in addition to OR7 in the
30 number of j, a substituent selected from the group consisting
of a halogen atom, a C1-6 alkyl group optionally substituted by
a halogen atom(s), and a C1-6 alkoxy group optionally
47
substituted by a halogen atom(s); or
Ra and Rb are joined to form an oxo group.
[0130]
Examples of the group protected by pseudo solid phase
protecting group (III) include a hydroxy group, 5 an amino group,
a carboxy group.
[0131]
Each aliphatic hydrocarbon group having not less than 10
carbon atoms for R5 in the formula (a2), the formula (a2’) and
the formula (a2”), and R7 10 for the formula (a3) is independently
preferably a group selected from a linear C10-40 alkyl group and
a linear C10-40 alkenyl group, more preferably a linear C10-40
alkyl group, further preferably a linear C10-30 alkyl group,
particularly preferably a linear C12-28 alkyl group, most
15 preferably a linear C14-26 alkyl group.
[0132]
Each linker for R5 in the formula (a2), the formula (a2’)
and the formula (a2”) and R7 for the formula (a3) is
independently preferably -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -
20 C(=O)NH- or -NHC(=O)-, more preferably -O-.
[0133]
R5 in the formula (a2), the formula (a2’) and the formula
(a2”) and R7 in the formula (a3) (namely, a hydrocarbon group
bonded via a single bond or a linker to an aliphatic
25 hydrocarbon group having not less than 10 carbon atoms) are
each independently preferably a group selected from a linear
C10-40 alkyl group and a linear C10-40 alkenyl group, more
preferably a linear C10-40 alkyl group, further preferably a
linear C10-30 alkyl group, particularly preferably a linear C12-28
30 alkyl group, most preferably a linear C14-26 alkyl group.
[0134]
Q in the formula (a2), the formula (a2’), the formula
(a2”) and the formula (a3) is preferably -O-.
[0135]
35 In the formula (III), a preferable embodiment of L
48
represented by the formula (a1) is a group wherein
L1 is a divalent C1-22 hydrocarbon group, or CH2-O-1,4-
phenylene-O-CH2; and
L2 is a single bond, or a group represented by
**C(=O)N(R2’)-R1’-N(R3)*** wherein ** shows the bonding 5 position to
L1, *** shows the bonding position to C=O, R1’ is a C1-6 alkylene
group, R2’ and R3 are each independently a hydrogen atom, or an
optionally substituted C1-6 alkyl group, or R2’ and R3 are
optionally joined to form an optionally substituted C1-6
10 alkylene bond.
[0136]
Another preferable embodiment of L represented by the
formula (a1) is a group wherein
L1 is a divalent C1-22 hydrocarbon group; and
15 L2 is a single bond.
[0137]
Another preferable embodiment of L represented by the
formula (a1) is a group wherein
L1 is an ethylene group; and
L2 is a group represented by **C(=O)N(R2’)-R1’-N(R3)*** 20
wherein ** shows the bonding position to L1, *** shows the
bonding position to C=O, R1’ is a C1-22 alkylene group, R2’ and R3
are each independently a hydrogen atom or a C1-22 alkyl group,
or R2’ and R3 are optionally joined to form a ring.
25 [0138]
Another preferable embodiment of L represented by the
formula (a1) is a group wherein
L1 is an ethylene group; and
L2 is a group represented by **C(=O)N(R2’)-R1’-N(R3)***
wherein ** shows the bonding position to L1, *** 30 shows the
bonding position to C=O, and N(R2’)-R1’-N(R3) moiety forms a
piperazinediyl group (e.g., 1,4-piperazinediyl group).
[0139]
Another preferable embodiment of L represented by the
35 formula (a1) is a group wherein
49
L1 is an ethylene group; and
L2 is a group represented by **C(=O)N(R2’)-R1’-N(R3)***
wherein ** shows the bonding position to L1, *** shows the
bonding position to C=O, R1’ is a pentylene group or a hexylene
group, and R2’ and R3 are each independently 5 a hydrogen atom or
a methyl group.
[0140]
A particularly preferable embodiment of L represented by
the formula (a1) is a succinyl group which is easily available
10 at a low cost.
[0141]
Now, L represented by the formula (a1’) in the formula
(III) is explained.
L1 in the formula (a1’) is preferably a divalent C6-10
15 aromatic hydrocarbon group, more preferably a phenylene group.
L2 in the formula (a1’) is preferably a single bond.
A preferable combination of L1 and L2 in the formula
(a1’) is a combination of L1 being a divalent C6-10 aromatic
hydrocarbon group, and L2 being a single bond. A more
20 preferable combination of L1 and L2 in the formula (a1’) is a
combination of L1 being a phenylene group, and L2 being a
single bond.
R8 and R9 in the formula (a1’) are each independently
preferably a C1-22 alkyl group, more preferably a C1-10 alkyl
25 group.
[0142]
A preferable embodiment of L represented by the formula
(a1’) is a group wherein
R8 and R9 are each independently a C1-22 alkyl group;
30 L1 is a divalent C6-10 aromatic hydrocarbon group; and
L2 is a single bond.
[0143]
Another preferable embodiment of L represented by the
formula (a1’) is a group wherein
R8 and R9 35 are each independently a C1-10 alkyl group;
50
L1 is a phenylene group; and
L2 is a single bond.
[0144]
Y in the formula (III) is a single bond, an oxygen atom,
or NR (R is a hydrogen atom, an alkyl group 5 or an aralkyl
group). When Y is a single bond, L is a single bond, Z is a
group represented by the formula (a2), R4 is a hydrogen atom,
and Ra and Rb are preferably joined to form an oxo group.
[0145]
10 Y in the formula (III) is preferably an oxygen atom, or
NR (R is a hydrogen atom, an alkyl group or an aralkyl group),
R is preferably a hydrogen atom, a C1-6 alkyl group or a C7-16
aralkyl group, more preferably a hydrogen atom, methyl, ethyl
or benzyl, particularly preferably a hydrogen atom. Y is
15 particularly preferably an oxygen atom.
[0146]
Z in the formula (III) is preferably a group represented
by the formula (a2) or the formula (a2’), more preferably a
group represented by the formula (a2). In the formula (a2), R4
20 is preferably a hydrogen atom. In the formula (a2), Ra and Rb
are each preferably a hydrogen atom.
[0147]
A preferable embodiment of Z represented by the formula
(a2) is a group wherein
25 Ra and Rb are each a hydrogen atom;
R4 is a hydrogen atom;
k is an integer of 1 - 3;
Q in the number of k are each -O-; and
R5 in the number of k are each independently a linear C10-
30 40 alkyl group.
When Z is this embodiment, it is more preferable that L
is a succinyl group, or a group represented by the formula
(a1’) (in the formula (a1’), R8 and R9 are each independently a
C1-10 alkyl group, L1 is a divalent phenylene group, and L2 is a
35 single bond), and Y is an oxygen atom, or L-Y is a single bond,
51
and it is further preferable that L is a succinyl group, or a
group represented by the formula (a1’) (in the formula (a1’),
R8 and R9 are each independently a C1-10 alkyl group, L1 is a
divalent phenylene group, and L2 is a single bond), and Y is an
5 oxygen atom.
[0148]
Another preferable embodiment of Z represented by the
formula (a2) is a group wherein
Ra and Rb are each a hydrogen atom;
R4 10 is a hydrogen atom;
k is an integer of 1 - 3;
Q in the number of k are each -O-; and
R5 in the number of k are each independently a benzyl group
bonded via -O- to 1 to 3 linear C10-40 alkyl groups, or a
15 cyclohexylmethyl group bonded via -O- to 1 to 3 linear C10-40
alkyl groups; and
ring A optionally further has, in addition to QR5 in the number
of k, substituent(s) selected from the group consisting of a
halogen atom, a C1-6 alkyl group optionally substituted by a
20 halogen atom, and a C1-6 alkoxy group optionally substituted by
a halogen atom.
When Z is this embodiment, it is more preferable that L
is a succinyl group, or a group represented by the formula
(a1’) (in the formula (a1’), R8 and R9 are each independently a
25 C1-10 alkyl group, L1 is a divalent phenylene group, and L2 is a
single bond), and Y is an oxygen atom, or L-Y is a single bond,
and it is further preferable that L is a succinyl group, or a
group represented by the formula (a1’) (in the formula (a1’),
R8 and R9 are each independently a C1-10 alkyl group, L1 is a
30 divalent phenylene group, and L2 is a single bond), and Y is an
oxygen atom.
[0149]
Another preferable embodiment of Z represented by the
formula (a2) is a group wherein
Ra and R4 35 are each a hydrogen atom; and
52
Rb is a group represented by the formula (a3) wherein *
is a bonding position, j is an integer of 0 - 3, Q in the
number of j is -O-, R7 in the number of j are each
independently a C10-40 alkyl group, R6 is a hydrogen atom.
When Z is this embodiment, it is more 5 preferable that L
is a succinyl group, or a group represented by the formula
(a1’) (in the formula (a1’), R8 and R9 are each independently a
C1-10 alkyl group, L1 is a divalent phenylene group, and L2 is a
single bond), and Y is an oxygen atom, or L-Y is a single bond,
10 and it is further preferable that L is a succinyl group, or a
group represented by the formula (a1’) (in the formula (a1’),
R8 and R9 are each independently a C1-10 alkyl group, L1 is a
divalent phenylene group, and L2 is a single bond), and Y is an
oxygen atom.
15 [0150]
A still another preferable embodiment of Z represented by
the formula (a2) is a group wherein
Ra is a hydrogen atom; and
Rb is a group represented by the formula (a3) wherein *
20 is a bonding position, j is an integer of 0 - 3, Q in the
number of j is -O-, R7 in the number of j are each
independently a linear C10-40 alkyl group, and R6 is joined with
R4 of ring A to form a single bond or -O-, whereby ring A and
ring C are joined to form a fluorene ring or a xanthene ring.
25 When Z is this embodiment, it is more preferable that L
is a succinyl group, or a group represented by the formula
(a1’) (in the formula (a1’), R8 and R9 are each independently a
C1-10 alkyl group, L1 is a divalent phenylene group, and L2 is a
single bond), and Y is an oxygen atom, or L-Y is a single bond,
30 and it is further preferable that L is a succinyl group, or a
group represented by the formula (a1’) (in the formula (a1’),
R8 and R9 are each independently a C1-10 alkyl group, L1 is a
divalent phenylene group, and L2 is a single bond), and Y is an
oxygen atom.
35 [0151]
53
Another preferable embodiment of Z represented by the
formula (a2) is a group wherein
Ra and Rb are joined to form an oxo group;
R4 is a hydrogen atom;
k is an 5 integer of 1 - 3;
Q in the number of k is -O-; and
R5 in the number of k are each independently linear C10-40
alkyl group.
When Z is this embodiment, it is more preferable that L-Y
10 is a single bond or a succinyl-1,4-piperazinediyl group, and it
is further preferable that L-Y is a single bond.
[0152]
Another preferable embodiment of Z represented by the
formula (a2) is a group wherein
15 Ra and Rb are joined to form an oxo group;
R4 is a hydrogen atom;
k is an integer of 1 - 3;
Q in the number of k are each -O-; and
R5 in the number of k are each independently a benzyl group
20 bonded via -O- to 1 to 3 linear C10-40 alkyl groups, or a
cyclohexylmethyl group bonded via -O- to 1 to 3 linear C10-40
alkyl groups; and
ring A optionally further has, in addition to QR5 in the number
of k, substituent(s) selected from the group consisting of a
25 halogen atom, a C1-6 alkyl group optionally substituted by a
halogen atom, and a C1-6 alkoxy group optionally substituted by
a halogen atom.
When Z is this embodiment, it is more preferable that L-Y
is a single bond or a succinyl-1,4-piperazinediyl group, and it
30 is further preferable that L-Y is a single bond.
[0153]
The pseudo solid phase protecting group (III) is
preferably a group not removed under acidic conditions but
removed under basic conditions.
35 [0154]
54
Specific examples 1 of pseudo solid phase protecting
group (III) having a linear aliphatic hydrocarbon group having
not less than 10 carbon atoms, which is for protecting a
hydroxy group include
groups wherein L is a succinyl 5 group, or a group
represented by the formula (a1’) (the formula (a1’) wherein R8
and R9 are each independently a C1-10 alkyl group, L1 is a
divalent phenylene group, and L2 is a single bond), and Y-Z is
a 3,4,5-tri(octadecyloxy)benzyloxy group, 2,4-
10 di(octadecyloxy)benzyloxy group, 2,4-di(docosyloxy)benzyloxy
group, 3,5-di(docosyloxy)benzyloxy group, 3,5-bis[3’,4’,5’-
tri(octadecyloxy)benzyloxy]benzyloxy group, 3,4,5-
tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzyloxy group,
3,4,5-tri(octadecyloxy)benzylamino group, 2,4-
15 di(docosyloxy)benzylamino group, 3,5-di(docosyloxy)benzylamino
group, bis(4-docosyloxyphenyl)methylamino group, 4-methoxy-2-
[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino group, 4-
methoxy-2-[3’,4’,5’-
tri(octadecyloxy)cyclohexylmethyloxy]benzylamino group, 2,4-
20 bis(dodecyloxy)benzylamino group, phenyl(2,3,4-
tri(octadecyloxy)phenyl)methylamino group, bis[4-(12-
docosyloxydodecyloxy)phenyl]methylamino group, 3,5-
bis[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino group,
3,4,5-tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino
25 group, 3,4,5-tri(octadecyloxy)cyclohexylmethyloxy group, 3,5-
di(docosyloxy)cyclohexylmethyloxy group, 3,5-bis[3’,4’,5’-
tri(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethyloxy group,
3,4,5-tris[3’,4’,5’-
tri(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethyloxy group,
30 3,4,5-tri(octadecyloxy)cyclohexylmethylamino group, 2,4-
di(docosyloxy)cyclohexylmethylamino group, 3,5-
di(docosyloxy)cyclohexylmethylamino group, bis(4-
docosyloxyphenyl)methylamino group, 4-methoxy-2-[3’,4’,5’-
tri(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino
35 group, 4-methoxy-2-[3’,4’,5’-
55
tri(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino
group, 2,4-bis(dodecyloxy)cyclohexylmethylamino group,
phenyl(2,3,4-tri(octadecyloxy)phenyl)methylamino group, bis[4-
(12-docosyloxydodecyloxy)phenyl]methylamino group, 3,5-
5 bis[3’,4’,5’-
tri(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino
group, or 3,4,5-tris[3’,4’,5’-
tri(octadecyloxy)cyclohexylmethyloxy]cyclohexylmethylamino
group,
10 groups wherein
L-Y is a succinyl-1,4-piperazinediyl group, and Z is a
3,4,5-tri(octadecyloxy)benzoyl group, 2,4-
di(octadecyloxy)benzoyl group, 2,4-di(docosyloxy)benzoyl group,
3,5-di(docosyloxy)benzoyl group, 3,5-bis[3’,4’,5’-
15 tri(octadecyloxy)benzyloxy]benzoyl group, or 3,4,5-
tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzoyl group, and
groups wherein
L-Y is a single bond, and Z is a (3,5-
didocosyloxyphenyl)diphenylmethyl group or bis(4-
20 chlorophenyl)(3,5-didocosyloxyphenyl)methyl group.
[0155]
Specific examples 2 of pseudo solid phase protecting
group (III) having a linear aliphatic hydrocarbon group having
not less than 10 carbon atoms, which is for protecting an amino
25 group or an imino group include groups wherein L-Y is a single
bond, and Z is a 3,4,5-tri(octadecyloxy)benzoyl group, 2,4-
di(octadecyloxy)benzoyl group, 2,4-di(docosyloxy)benzoyl group,
3,5-di(docosyloxy)benzoyl group, 3,5-bis[3’,4’,5’-
tri(octadecyloxy)benzyloxy]benzoyl group, 3,4,5-tris[3’,4’,5’-
30 tri(octadecyloxy)benzyloxy]benzoyl group, (3,5-
didocosyloxyphenyl)diphenylmethyl group, or bis(4-
chlorophenyl)(3,5-didocosyloxyphenyl)methyl group.
[0156]
Specific examples 3 of pseudo solid phase protecting
35 group (III) having a linear aliphatic hydrocarbon group having
56
not less than 10 carbon atoms, which is for protecting a
carboxy group include groups wherein L-Y is a single bond, and
Z is a 3,4,5-tri(octadecyloxy)benzyl group, 2,4-
di(octadecyloxy)benzyl group, 2,4-di(docosyloxy)benzyl group,
3,5-di(docosyloxy)benzyl group, 5 3,5-bis[3’,4’,5’-
tri(octadecyloxy)benzyloxy]benzyl group, 3,4,5-tris[3’,4’,5’-
tri(octadecyloxy)benzyloxy]benzyl group, (3,5-
didocosyloxyphenyl)diphenylmethyl group, or bis(4-
chlorophenyl)(3,5-didocosyloxyphenyl)methyl group.
10 [0157]
Specific examples 4 of pseudo solid phase protecting
group (III) having a branched chain aliphatic hydrocarbon group
having not less than 10 carbon atoms, which is for protecting a
hydroxy group include groups wherein L is a succinyl group, or
15 a group represented by the formula (a1’) (in the formula (a1’),
R8 and R9 are each independently a C1-10 alkyl group, L1 is a
divalent phenylene group, and L2 is a single bond), and Y-Z is
a 4-(2’,3’-dihydrophytyloxy)benzyloxy group, 4-(3,7,11-
trimethyldodecyloxy)benzyloxy group, 4-[2,2,4,8,10,10-
20 hexamethyl-5-dodecanoylamino]benzyloxy group, 4-(2’,3’-
dihydrophytyloxy)-2-methylbenzyloxy group, 4-(2’,3’-
dihydrophytyloxy)-2-methoxybenzyloxy group, 2,4-di(2’,3’-
dihydrophytyloxy)benzyloxy group, 3,5-di(2’,3’-
dihydrophytyloxy)benzyloxy group, 2-[3’,4’,5’-tri(2”,3”-
25 dihydrophytyloxy)benzyloxy]-4-methoxybenzyloxy group, 3,4,5-
tri(2’,3’-dihydrophytyloxy)benzyloxy group, 3,4,5-tris[3,4,5-
tri(2’,3’-dihydrophytyloxy)benzyloxy]benzyloxy group, 2,4-di(2-
decyltetradecyloxy)benzyloxy group, 4-(2’,3’-
dihydrophytyloxy)benzylamino group, 4-(3,7,11-
30 trimethyldodecyloxy)benzylamino group, 4-[2,2,4,8,10,10-
hexamethyl-5-dodecanoylamino]benzylamino group, 4-(2’,3’-
dihydrophytyloxy)-2-methylbenzylamino group, 4-(2’,3’-
dihydrophytyloxy)-2-methoxybenzylamino group, 2,4-di(2’,3’-
dihydrophytyloxy)benzylamino group, 3,5-di(2’,3’-
35 dihydrophytyloxy)benzylamino group, 2-[3’,4’,5’-tri(2”,3”-
57
dihydrophytyloxy)benzyloxy]-4-methoxybenzylamino group, 3,4,5-
tri(2’,3’-dihydrophytyloxy)benzylamino group, 3,4,5-tris[3,4,5-
tri(2’,3’-dihydrophytyloxy)benzyloxy]benzylamino group, {[(2-
chloro-5-(2’,3’-dihydrophytyloxy)phenyl)]phenylmethyl}amino
group, 5 {[2,3,4-tri(2’,3’-
dihydrophytyloxy)phenyl]phenylmethyl}amino group, or {bis[4-
(2’,3’-dihydrophytyloxy)phenyl]methyl}amino group.
Here, 2,3-dihydrophytyl means 3,7,11,15-
tetramethylhexadecyl. The same applies to 2’,3’-dihydrophytyl
10 and 2”,3”-dihydrophytyl.
[0158]
Specific examples 5 of pseudo solid phase protecting
group (III) having a branched chain aliphatic hydrocarbon group
having not less than 10 carbon atoms, which is for protecting
15 an amino group or an imino group include groups wherein L-Y is
a single bond, and Z is a 4-(2’,3’-dihydrophytyloxy)benzoyl
group, 4-(3,7,11-trimethyldodecyloxy)benzoyl group, 4-
[2,2,4,8,10,10-hexamethyl-5-dodecanoylamino]benzoyl group, 4-
(2’,3’-dihydrophytyloxy)-2-methylbenzoyl group, 4-(2’,3’-
20 dihydrophytyloxy)-2-methoxybenzoyl group, 2,4-di(2’,3’-
dihydrophytyloxy)benzoyl group, 3,5-di(2’,3’-
dihydrophytyloxy)benzoyl group, 2-[3’,4’,5’-tri(2”,3”-
dihydrophytyloxy)benzyloxy]-4-methoxybenzoyl group, 3,4,5-
tri(2’,3’-dihydrophytyloxy)benzoyl group, 3,4,5-tris[3,4,5-
25 tri(2’,3’-dihydrophytyloxy)benzyloxy]benzoyl group, or 2,4-
di(2-decyltetradecyloxy)benzoyl group.
[0159]
Specific examples 6 of pseudo solid phase protecting
group (III) having a branched chain aliphatic hydrocarbon group
30 having not less than 10 carbon atoms, which is for protecting a
carboxy group include
groups wherein
L-Y is a single bond, and Z is a 4-(2’,3’-
dihydrophytyloxy)benzyl group, 4-(3,7,11-
35 trimethyldodecyloxy)benzyl group, 4-[2,2,4,8,10,10-hexamethyl58
5-dodecanoylamino]benzyl group, 4-(2’,3’-dihydrophytyloxy)-2-
methylbenzyl group, 4-(2’,3’-dihydrophytyloxy)-2-methoxybenzyl
group, 2,4-di(2’,3’-dihydrophytyloxy)benzyl group, 3,5-
di(2’,3’-dihydrophytyloxy)benzyl group, 2-[3’,4’,5’-tri(2”,3”-
dihydrophytyloxy)benzyloxy]-4-methoxybenzyl 5 group, 3,4,5-
tri(2’,3’-dihydrophytyloxy)benzyl group, 3,4,5-tris[3,4,5-
tri(2’,3’-dihydrophytyloxy)benzyloxy]benzyl group, or 2,4-di(2-
decyltetradecyloxy)benzyl group.
[0160]
10 The pseudo solid phase protecting group (III) is
preferably any of the aforementioned Specific examples 1 -
Specific examples 6, more preferably Specific examples 1 -
Specific examples 3, of pseudo solid phase protecting group
(III).
15 [0161]
The pseudo solid phase protecting group (III) is still
more preferably
a group wherein L is a succinyl group or a group
represented by the formula (a1’) (the formula (a1’) wherein R8
and R9 20 are each independently a C1-10 alkyl group, L1 is a
divalent phenylene group, L2 is a single bond), and Y-Z is a
3,4,5-tri(octadecyloxy)benzyloxy group, 2,4-
di(octadecyloxy)benzyloxy group, 2,4-di(docosyloxy)benzyloxy
group, 3,5-di(docosyloxy)benzyloxy group, 3,5-bis[3’,4’,5’-
25 tri(octadecyloxy)benzyloxy]benzyloxy group, 3,4,5-
tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzyloxy group,
3,4,5-tri(octadecyloxy)benzylamino group, 2,4-
di(docosyloxy)benzylamino group, 3,5-di(docosyloxy)benzylamino
group, di(4-docosyloxyphenyl)methylamino group, 4-methoxy-2-
30 [3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino group, 4-
methoxy-2-[3’,4’,5’-
tri(octadecyloxy)cyclohexylmethyloxy]benzylamino group, 2,4-
di(dodecyloxy)benzylamino group, phenyl(2,3,4-
tri(octadecyloxy)phenyl)methylamino group, bis[4-(12-
35 docosyloxydodecyloxy)phenyl]methylamino group, 3,5-
59
bis[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino group, or
3,4,5-tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino
group,
a group wherein L-Y is a succinyl-1,4-piperazinediyl
group, and Z is a 3,4,5-tri(octadecyloxy)5 benzoyl group, 2,4-
di(octadecyloxy)benzoyl group, 2,4-di(docosyloxy)benzoyl group,
3,5-di(docosyloxy)benzoyl group, 3,5-bis[3’,4’,5’-
tri(octadecyloxy)benzyloxy]benzoyl group, or 3,4,5-
tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzoyl group, or
10 a group wherein L-Y is a single bond, and Z is a 3,4,5-
tri(octadecyloxy)benzoyl group, 2,4-di(octadecyloxy)benzoyl
group, 2,4-di(docosyloxy)benzoyl group, 3,5-
di(docosyloxy)benzoyl group, 3,5-bis[3’,4’,5’-
tri(octadecyloxy)benzyloxy]benzoyl group, or 3,4,5-
15 tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzoyl group.
[0162]
The pseudo solid phase protecting group (III) is further
preferably
a group wherein L is a succinyl group, or a group
represented by the formula (a1’) (the formula (a1’) wherein R8 20
and R9 are each independently a C1-10 alkyl group, L1 is a
divalent phenylene group, L2 is a single bond), and Y-Z is a
3,4,5-tri(octadecyloxy)benzyloxy group, 3,5-
di(docosyloxy)benzyloxy group, 3,5-bis[3’,4’,5’-
25 tri(octadecyloxy)benzyloxy]benzyloxy group, 3,4,5-
tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzyloxy group,
3,4,5-tri(octadecyloxy)benzylamino group, 2,4-
di(docosyloxy)benzylamino group, 3,5-di(docosyloxy)benzylamino
group, di(4-docosyloxyphenyl)methylamino group, 4-methoxy-2-
30 [3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino group, 4-
methoxy-2-[3’,4’,5’-
tri(octadecyloxy)cyclohexylmethyloxy]benzylamino group, 2,4-
di(dodecyloxy)benzylamino group, phenyl(2,3,4-
tri(octadecyloxy)phenyl)methylamino group, bis[4-(12-
35 docosyloxydodecyloxy)phenyl]methylamino group, 3,5-
60
bis[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino group, or
3,4,5-tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino
group, or
a group wherein L-Y is a single bond, and Z is a 3,4,5-
tri(octadecyloxy)5 benzoyl group.
[0163]
The pseudo solid phase protecting group (III) is
particularly preferably
a group wherein L is a succinyl group, and Y-Z is a
10 3,4,5-tri(octadecyloxy)benzyloxy group, 3,5-
di(docosyloxy)benzyloxy group, 3,5-bis[3’,4’,5’-
tri(octadecyloxy)benzyloxy]benzyloxy group, 3,4,5-
tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzyloxy group,
3,4,5-tri(octadecyloxy)benzylamino group, 2,4-
15 di(docosyloxy)benzylamino group, 3,5-di(docosyloxy)benzylamino
group, di(4-docosyloxyphenyl)methylamino group, 4-methoxy-2-
[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino group, 4-
methoxy-2-[3’,4’,5’-
tri(octadecyloxy)cyclohexylmethyloxy]benzylamino group, 2,4-
20 di(dodecyloxy)benzylamino group, phenyl(2,3,4-
tri(octadecyloxy)phenyl)methylamino group, bis[4-(12-
docosyloxydodecyloxy)phenyl]methylamino group, 3,5-
bis[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino group, or
3,4,5-tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino
25 group, or
a group wherein L-Y is a single bond, and Z is a 3,4,5-
tri(octadecyloxy)benzoyl group.
[0164]
The pseudo solid phase protecting group (III) is most
30 preferably a group wherein L is a succinyl group, and Y-Z is a
3,4,5-tri(octadecyloxy)benzyloxy group, 3,5-
di(docosyloxy)benzyloxy group, 3,5-bis[3’,4’,5’-
tri(octadecyloxy)benzyloxy]benzyloxy group, 3,4,5-
tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzyloxy group,
35 3,4,5-tri(octadecyloxy)benzylamino group, 2,4-
61
di(docosyloxy)benzylamino group, 3,5-di(docosyloxy)benzylamino
group, di(4-docosyloxyphenyl)methylamino group, 4-methoxy-2-
[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino group, 4-
methoxy-2-[3’,4’,5’-
tri(octadecyloxy)cyclohexylmethyloxy]benzylamino 5 group, 2,4-
di(dodecyloxy)benzylamino group, phenyl(2,3,4-
tri(octadecyloxy)phenyl)methylamino group, bis[4-(12-
docosyloxydodecyloxy)phenyl]methylamino group, 3,5-
bis[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino group, or
10 3,4,5-tris[3’,4’,5’-tri(octadecyloxy)benzyloxy]benzylamino
group.
[0165]
A compound used for forming a pseudo solid phase
protecting group can be produced by a method known per se (e.g.,
15 the method described in the aforementioned patent document 1 or
2, esterification, silylation etc.) or a method analogous
thereto. For example, an alcohol compound or an amine compound
represented by the formula: Z-Y-H used for forming a pseudo
solid phase protecting group can be produced by the following
20 steps or steps analogous thereto.
[0166]
62
[0167]
wherein Q’ is -O-, -S-, -C(=O)O- or -NH-, Rg is a hydrogen atom,
an ORh group (wherein Rh is an alkyl group such as C1-6 alkyl
group and the like, an aralkyl group such as 5 benzyl group and
the like, and the like) or a group represented by the formula
(a3):
[0168]
10
[0169]
wherein each symbol is as defined above, Y1 is a leaving group
63
such as a halogen atom and the like, and other symbols are as
defined above.
[0170]
step (a)
In this step, R5 is introduced into Q’H 5 (wherein Q’ is -
O-, -S-, -C(=O)O- or -NH-) in a compound represented by the
formula (X) to produce a compound represented by the formula
(X-a). When Q’ is -O-, -S- or -NH-, this reaction is performed
in a solvent that does not influence the reaction, in the
10 presence or absence of a base and using halide (chloride,
bromide or iodide) corresponding to R5, carboxylic acid or acid
halide corresponding to R5 or an alkylsulfonyloxylation product
(e.g., methanesulfonyloxylation product etc.) or an
arylsulfonyloxylation product (e.g., p-toluenesulfonyloxylation
product etc.) corresponding to R515 . When Q’ is -O-, this
reaction can also be performed under Mitsunobu reaction
conditions in which compound (X) and hydroxide corresponding to
R5 are reacted in the presence of triphenylphosphine and
diisopropyl azodicarboxylate. Furthermore, when Q’ is -C(=O)O-,
20 for example, compound (X-a) can be synthesized by reacting
compound (X) with amine or hydroxide corresponding to R5 in the
presence of a condensing agent.
[0171]
Examples of the base include alkali metal salts such as
25 sodium carbonate, sodium hydrogen carbonate, potassium
carbonate, potassium tert-butoxide and the like; alkali metal
hydrides such as sodium hydride, potassium hydride and the
like; amines such as pyridine, triethylamine, N,Ndimethylaniline,
1,8-diazabicyclo[5.4.0]-7-undecene and the
30 like, and the like, of which sodium carbonate, potassium
carbonate, sodium hydride and the like are preferable.
[0172]
Examples of the solvent include aromatic hydrocarbons
such as toluene, xylene and the like; ethers such as
35 tetrahydrofuran, dioxane and the like; amides such as
64
dimethylformamide, dimethylacetamide and the like; halogenated
hydrocarbons such as chloroform, dichloromethane and the like;
nitriles such as acetonitrile and the like, N-methylpyrrolidone
and the like, or a mixture thereof, of which dimethylformamide,
tetrahydrofuran, toluene, N-methylpyrrolidone 5 and the like are
preferable.
[0173]
The reaction temperature is preferably 50 - 150C, more
preferably 60 - 130C. The reaction time is preferably 2 - 30
10 hr, more preferably 3 - 10 hr.
[0174]
step (b)
In this step, compound (X-a) is reduced to produce a
compound represented by the formula (XI-a). This reduction
15 reaction can be performed by a method using a reducing agent.
[0175]
Examples of the reducing agent to be used for this
reduction reaction include metal hydrides (sodium borohydride,
lithium borohydride, sodium cyanoborohydride, sodium
20 triacetoxyborohydride, dibutylaluminum hydride, aluminum
hydride, lithium aluminum hydride etc.) and the like, of which
sodium borohydride, dibutylaluminum hydride and the like are
preferable.
[0176]
25 This reaction is performed in a solvent that does not
influence the reaction. Examples of the solvent include
alcohols such as methanol, ethanol and the like; ethers such as
diethyl ether, tetrahydrofuran, dioxane and the like; aromatic
hydrocarbons such as toluene, xylene and the like; or a mixture
30 thereof, of which tetrahydrofuran, toluene and the like are
preferable. The reaction temperature is preferably 0 - 100C,
more preferably 30 - 70C. The reaction time is preferably 1 -
24 hr, more preferably 2 - 5 hr.
[0177]
35 step (c)
65
In this step, compound (X-a) wherein Rg is not a hydrogen
atom or ORh group is reduced by a method similar to the abovementioned
step (b).
[0178]
5 step (d-1)
In this step, compound (X-a) wherein Rg is a hydrogen
atom is oximated to produce a compound represented by the
formula (XI’-a).
[0179]
10 The oximation reaction is performed by reacting compound
(X-a) with an acid addition salt of hydroxylamine in a solvent
that does not influence the reaction, in the presence of a base.
[0180]
Examples of the acid addition salt of hydroxylamine
15 include salts of mineral acids such as hydrochloride, sulfate,
nitrate salt and the like, organic acid salts such as acetate,
trifluoroacetate, methanesulfonate, trifluoromethanesulfonate,
p-toluenesulfonate and the like, and the like, and
hydrochloride is particularly preferable.
20 [0181]
Examples of the base include alkali metal salts such as
potassium hydroxide, sodium hydroxide, sodium hydrogen
carbonate, potassium carbonate and the like; organic amines
such as pyridine, triethylamine, diisopropylethylamine, N,N25
dimethylaniline, 1,8-diazabicyclo[5.4.0]undeca-7-en and the
like, and the like, of which triethylamine,
diisopropylethylamine and the like are preferable.
[0182]
Examples of the solvent include halogenated hydrocarbons
30 such as chloroform, dichloromethane and the like; aromatic
hydrocarbons such as toluene, xylene and the like; ethers such
as tetrahydrofuran, dioxane and the like; and/or a mixture
thereof, of which dichloromethane, chloroform, toluene and the
like are preferable. The reaction temperature is preferably 10
35 - 100C, more preferably 20 - 60C. The reaction time is
66
preferably 0.5 - 30 hr, more preferably 2 - 20 hr.
[0183]
step (d-2)
In this step, compound (XI’-a) is reduced by catalytic
hydrogenation reaction in the presence of a metal 5 catalyst such
as palladium-carbon, Raney-nickel and the like, or reducing
agent such as metal hydride and the like similar to that in the
aforementioned step (b) to produce a compound represented by
the formula (XI-b).
10 [0184]
Compound (XI-b) can also be produced by step (d-3), step
(d-4) and step (d-5).
[0185]
step (d-3)
15 In this step, compound (XI-a) is halogenated using, for
example, a chlorinating agent such as acetyl chloride, thionyl
chloride and the like, or a brominating agent such as acetyl
bromide, phosphorus tribromide, diphenylphosphine/bromine and
the like to produce a compound represented by the formula (XI’-
20 b).
[0186]
Examples of the solvent include halogenated hydrocarbons
such as chloroform, dichloromethane and the like; aromatic
hydrocarbons such as toluene, xylene and the like; ethers such
25 as tetrahydrofuran, dioxane and the like; and a mixture thereof,
of which chloroform, tetrahydrofuran, toluene and the like are
preferable. The reaction temperature is preferably 10 - 150C,
more preferably 30 - 80C. The reaction time is preferably 0.5
- 30 hr, more preferably 2 - 20 hr.
30 [0187]
step (d-4)
In this step, compound (XI’-b) is azidated using an
azidating agent such as sodium azide and the like to produce a
compound represented by the formula (XI’-c). This reaction is
35 performed by reacting compound (XI’-b) with an azidating agent
67
in a solvent that does not influence the reaction.
[0188]
Examples of the solvent include halogenated hydrocarbons
such as chloroform, dichloromethane and the like; aromatic
hydrocarbons such as toluene, xylene and the 5 like; ethers such
as tetrahydrofuran, dioxane and the like; amides such as N,Ndimethylformamide
and the like; and a mixture thereof, of which
chloroform, N,N-dimethylformamide and the like are preferable.
The reaction temperature is preferably 10 - 150C, more
10 preferably 20 - 100C. The reaction time is preferably 0.5 -
30 hr, more preferably 2 - 20 hr.
[0189]
step (d-5)
In this step, compound (XI’-c) is aminated to produce
15 compound (XI-b). This reaction is performed by reacting
compound (XI’-c) with triphenylphosphine in a solvent that does
not influence the reaction, in the presence of water, or by
catalytic hydrogenation reduction.
[0190]
20 The amount of triphenylphosphine to be used is preferably
1 - 10 mol, particularly preferably 1 - 5 mol, per 1 mol of
compound (XI’-c). The amount of water to be used is preferably
1 - 10 mol, particularly preferably 1 - 5 mol, per 1 mol of
compound (XI’-c).
25 [0191]
Examples of the solvent include aromatic hydrocarbons
such as toluene, xylene and the like; ethers such as
tetrahydrofuran, dioxane and the like; and a mixture thereof,
of which toluene, tetrahydrofuran and the like are preferable.
30 The reaction temperature is preferably 10 - 150C, more
preferably 20 - 100C. The reaction time is preferably 0.5 -
30 hr, more preferably 2 - for 20 hr.
[0192]
step (d-6)
35 In this step, compound (XI’-b) is reacted with RNH2 (R is
68
as defined above) to produce a compound represented by the
formula (XI-c) wherein Y is a -NHR group. In this step,
compound (XI’-b) is reacted with amine represented by R-NH2 in
a solvent that does not influence the reaction and, where
necessary, in the presence of, for example, 5 a base such as
tertiary amine and the like (e.g., triethylamine,
diisopropylethylamine and the like).
[0193]
Examples of the solvent include aromatic hydrocarbons
10 such as toluene, xylene and the like; ethers such as
tetrahydrofuran, dioxane and the like; and, halogenated
hydrocarbons such as chloroform, dichloromethane and the like,
and a mixture thereof, of which toluene, tetrahydrofuran,
chloroform and the like are preferable. The reaction
15 temperature is preferably 10 - 100C, more preferably 20 - 60C.
The reaction time is preferably 0.5 - 30 hr, more preferably 2
- 20 hr.
[0194]
step (d-7)
20 In this step, compound (XI-d) is reacted with a compound
having a -CONH2 group or a -OCONH2 group, and treated with a
base to produce compound (XI-e). The reaction of compound (XId)
and a compound having a -CONH2 group or a -OCONH2 group is
performed in a solvent that does not influence the reaction,
25 under an acid catalyst.
[0195]
Examples of the acid catalyst include methanesulfonic
acid, trifluoromethanesulfonic acid, toluenesulfonic acid and
the like, of which methanesulfonic acid, toluenesulfonic acid
30 are preferable. The amount of the acid catalyst to be used is
preferably 0.05 - 0.5 mol, particularly preferably 0.1 - 0.3
mol, per 1 mol of compound (XI-d).
[0196]
Examples of the compound having a -CONH2 group or a -
35 OCONH2 group include Fmoc-NH2, HCONH2, CF3CONH2, AcNH2, EtOCONH2,
69
Cbz-NH2 and the like, of which Fmoc-NH2, EtOCONH2 and the like
are preferable. As used herein, “Fmoc-” means a 9-
fluorenylmethoxycarbonyl group (hereinafter to be also referred
to as Fmoc group), and “Cbz-” means a benzyloxycarbonyl group
(hereinafter to be also referred to 5 as Cbz group).
[0197]
As the reagent to be used as a material for step (a)
[i.e., hydroxide, halide, alkylsulfonyloxylation product (e.g.,
methanesulfonyloxylation product etc.) or an
10 arylsulfonyloxylation product (e.g., p-toluenesulfonyloxylation
product etc. corresponding to R5), hereinafter to be
abbreviated as “reagent of step (a)”], a commercially available
product can be used. In addition, the reagent of step (a) can
be produced, for example, by
15 (1) halogenation, alkylsulfonyloxylation or
arylsulfonyloxylation of hydroxide corresponding to R5, or
(2) reduction reaction of unsaturated hydroxide corresponding
to R5 (e.g., catalytic hydrogenation reaction etc. in the
presence of a metal catalyst such as platinum-carbon (Pt/C),
20 palladium-carbon (Pd/C), rhodium-carbon (Rh/C), Raney-nickel
and the like), and subsequent halogenation,
alkylsulfonyloxylation or arylsulfonyloxylation.
[0198]
Examples of the reagent for conversion of a hydroxy group
25 to a leaving group in the production of the reagent of step (a)
include, in addition to halogenating agents such as
chlorinating agents (e.g., thionyl chloride, Nchlorosuccinimide
(NCS) and the like), brominating agents (e.g.,
hydrobromic acid, acetyl bromide, N-bromosuccinimide (NBS),
30 phosphorus tribromide, diphenylphosphine/bromine and the like)
and the like, alkylsulfonylating agents (e.g., methanesulfonyl
chloride, trifluoromethanesulfonyl chloride and the like),
arylsulfonylating agents (e.g., benzenesulfonyl chloride, ptoluenesulfonyl
chloride and the like), and the like, of which
35 halogenating agents such as thionyl chloride, hydrobromic acid
70
and the like are preferable.
[0199]
This reaction is performed in a solvent that does not
influence the reaction, and examples of the solvent include
water; halogenated hydrocarbons such 5 as chloroform,
dichloromethane and the like; aromatic hydrocarbons such as
benzene, toluene, xylene and the like; nitriles such as
acetonitrile, propionitrile and the like; ethers such as
tetrahydrofuran, 1,4-dioxane, diethyl ether and the like, of
10 which water, halogenated hydrocarbons such as chloroform and
the like are preferable. The reaction temperature is
preferably 10 - 120C, more preferably 50 - 100C. The
reaction time is preferably 1 - 72 hr, more preferably 3 - 24
hr.
15 [0200]
A compound represented by the formula: Z-Y-H wherein Q’
is a single bond can also be produced by, for example, the
following method. That is, introduction of R5 on the benzene
ring can be performed by
20 (1) Friedel-Crafts reaction using halide (chloride, bromide, or
iodide) corresponding to R5, or carboxylic acid or acid halide
corresponding to R5,
(2) a method including subjecting a compound corresponding to
the above-mentioned compound (X) (compound wherein Q’H is
25 replaced with -CHO group) to carbon homologation by a Wittig
reaction, followed by catalytic hydrogenation and the like, or
(3) conventionally-used organic synthesis reaction such as
cross coupling using a metal catalyst and the like.
[0201]
30 The reaction reagent and the like in the above-mentioned
respective schemes are shown for convenience, and can be
appropriately changed within the range of the above-mentioned
definition.
[0202]
35 [Organic compound protected by the pseudo solid phase
71
protecting group of the present invention]
The organic compound protected by an organic group having
one or more aliphatic hydrocarbon groups having not less than
10 carbon atoms (the pseudo solid phase protecting group of the
present invention) is not particularly limited 5 as long as it
can be precipitated in a solvent, and an organic compound
having a functional group such as hydroxy group (-OH), sulfanyl
group (-SH), a carboxy group (-COOH), amino group (-NHR), imino
group (-NH-), carboxamide group (-CONHR”) (wherein R” is a
10 hydrogen atom or a hydrocarbon group) and the like, wherein the
functional group is protected by the pseudo solid phase
protecting group of the present invention (hereinafter
sometimes to be referred to as “protected organic compound”),
can be used. The protected organic compound may be further
15 protected by a protecting group different from the pseudo solid
phase protecting group (e.g., protecting group used for nucleic
acid synthesis). Examples of the protected organic compound
include nucleoside, nucleotide, amino acid, peptide and the
like disclosed in WO 2012/157723, WO 2012/157723, WO
20 2014/189142, JP-A-2010-116418, WO 2010/104169, WO 2010/113939,
WO 2011/078295, WO 2012/029794, JP-A-2009-185063, JP-A-2010-
275254 and the like.
[0203]
A preferable protected organic compound is nucleoside,
25 nucleotide or oligonucleotide optionally further protected by a
protecting group used for nucleic acid synthesis, or amino acid
or peptide optionally further protected by a protecting group
used for peptide synthesis. In the aforementioned nucleoside,
nucleotide or oligonucleotide, at least one group selected from
30 amino group and imino group of nucleic acid base, 2’- and 3’-
hydroxy groups of ribose residue, and 3’-hydroxy group of
deoxyribose residue is preferably protected by the pseudo solid
phase protecting group of the present invention. When the
aforementioned nucleoside, nucleotide or oligonucleotide is
35 morpholino nucleoside, morpholino nucleotide or morpholino
72
oligonucleotide, the 5’-hydroxy group of morpholine residue,
rather than the aforementioned hydroxy group, may be protected
by a pseudo solid phase protecting group.
[0204]
A more preferable protected organic 5 compound is a
nucleoside or oligonucleotide, wherein at least one group
selected from amino group and imino group of nucleic acid base,
2’- and 3’-hydroxy groups of ribose residue, 3’-hydroxy group
of deoxyribose residue, and 5’-hydroxy group of morpholine
10 residue is protected by the pseudo solid phase protecting group
of the present invention, and other group is optionally further
protected by a protecting group used for nucleic acid synthesis.
[0205]
A further preferable protected organic compound is a
15 nucleoside or oligonucleotide, wherein at least one group
selected from amino group and imino group of nucleic acid base,
2’- and 3’-hydroxy groups of ribose residue, and 3’-hydroxy
group of deoxyribose residue is protected by the pseudo solid
phase protecting group of the present invention, and other
20 group is optionally further protected by a protecting group
used for nucleic acid synthesis.
[0206]
A particularly preferable protected organic compound is a
nucleoside or oligonucleotide, wherein at least one group
25 selected from amino group and imino group of nucleic acid base,
3’-hydroxy group of ribose residue, and 3’-hydroxy group of
deoxyribose residue is protected by the pseudo solid phase
protecting group of the present invention, and other group is
optionally further protected by a protecting group used for
30 nucleic acid synthesis.
[0207]
The most preferable protected organic compound is a
nucleoside or oligonucleotide, wherein 3’-hydroxy group of
ribose residue or 3’-hydroxy group of deoxyribose residue is
35 protected by the pseudo solid phase protecting group of the
73
present invention, and other group is optionally further
protected by a protecting group used for nucleic acid synthesis.
[0208]
[Precipitation]
The present invention also provides 5 a method of
precipitating a protected organic compound in a solvent by
using the precipitation promoter of the present invention
(hereinafter sometimes to be referred to as “the precipitation
method of the present invention”), and a precipitation mixture
10 obtained by such method, which contains the precipitation
promoter and the protected organic compound. The
aforementioned solvent is preferably a solvent containing a
polar solvent, more preferably a mixed solvent of a polar
solvent and a nonpolar solvent.
15 [0209]
An organic compound protected by the pseudo solid phase
protecting group of the present invention shows improved
solubility in a nonpolar solvent as well as decreased
solubility in a polar solvent, due to a pseudo solid phase
20 protecting group which is a hydrophobic group. An organic
compound protected by such pseudo solid phase protecting group
is, for example, precipitated by adding a polar solvent to a
solution of the organic compound dissolved in a nonpolar
solvent, to achieve solid-liquid separation. The precipitation
25 promoter of the present invention can promote precipitation of
an organic compound protected by a pseudo solid phase
protecting group in a solvent, and can improve the recovery
rate thereof. In addition, a deprotected organic compound can
also be obtained by removing the pseudo solid phase protecting
30 group after obtaining the object product (i.e., organic
compound protected by pseudo solid phase protecting group).
[0210]
Examples of the polar solvent include alcohol solvents
such as methanol, ethanol, isopropanol and the like; nitrile
35 solvents such as acetonitrile, propionitrile and the like;
74
ketone solvents such as acetone, 2-butanone and the like; polar
ether solvents such as 1,4-dioxane, tetrahydrofuran and the
like; amide solvents such as dimethylformamide,
dimethylacetamide, N-methylpiperidone and the like, sulfoxide
solvents such as dimethyl sulfoxide and the 5 like. Only one
kind of polar solvent may be used, or two or more kinds thereof
may be used in combination. As the polar solvent, acetonitrile
is particularly preferable. From the aspect of precipitation
efficiency in the synthesis of oligonucleotide, the recovery
10 rate tends to increase when methanol is used as a polar solvent.
Since methanol inhibits coupling reaction in the
phosphoramidite method, methanol needs to be completely removed
from the precipitated oligonucleotide. However, drying takes a
long time and it is problematically difficult to evaluate
15 whether methanol remains in the system. On the other hand,
acetonitrile tends to decrease precipitation efficiency, but
does not inhibit coupling reaction in the phosphoramidite
method. Therefore, oligonucleotide precipitated using
acetonitrile does not require strict drying and evaluation of
20 residual solvent. Using the precipitation promoter of the
present invention, the object product can be precipitated and
recovered in a high yield even when acetonitrile is used.
[0211]
Examples of the nonpolar solvent include halogenated
25 solvents such as chloroform, dichloromethane, 1,2-
dichloroethane and the like; aromatic solvents such as benzene,
toluene, xylene, mesitylene and the like; ester solvents such
as ethyl acetate, isopropyl acetate and the like; aliphatic
solvents such as hexane, pentane, heptane, octane, nonane,
30 cyclohexane and the like; non-polar ether solvents such as
diethyl ether, cyclopentyl methyl ether, tert-butyl methyl
ether and the like. Only one kind of nonpolar solvent may be
used, or two or more kinds thereof may be used in combination.
As the nonpolar solvent, halogenated solvents, aromatic
35 solvents, ester solvents, aliphatic solvents, and a combination
75
of these are preferable; dichloromethane, chloroform, 1,2-
dichloroethane, benzene, toluene, xylene, mesitylene, hexane,
pentane, heptane, nonane, cyclohexane, ethyl acetate, isopropyl
acetate and a combination of these are more preferable;
chloroform, dichloromethane, toluene, and 5 a combination of
these are more preferable.
[0212]
To sufficiently precipitate a protected organic compound,
the amount of the precipitation promoter of the present
10 invention to be used is preferably not less than 0.1 molar

We claim:
1. A precipitation promoter for precipitating an organic
compound protected by an organic group having one or more
aliphatic hydrocarbon groups having not less 5 than 10 carbon
atoms in a solvent, which has one or more linear aliphatic
hydrocarbon groups having not less than 10 carbon atoms,
wherein the aliphatic hydrocarbon group in the precipitation
promoter has not less than 20 carbon atoms in total.
10
2. The precipitation promoter according to claim 1, wherein the
linear aliphatic hydrocarbon group having not less than 10
carbon atoms in the precipitation promoter is a group selected
from a linear C10-40 alkyl group and a linear C10-40 alkenyl group.
15
3. The precipitation promoter according to claim 1 or 2,
wherein the aliphatic hydrocarbon group having not less than 10
carbon atoms of the organic group is linear.
20 4. The precipitation promoter according to claim 1 or 2,
wherein the aliphatic hydrocarbon group having not less than 10
carbon atoms of the organic group is a group selected from a
linear C10-40 alkyl group and a linear C10-40 alkenyl group.
25 5. The precipitation promoter according to any one of claims 1
to 4, which is (1) an organic compound having one or more
structures represented by the formula (G):
wherein
each R1 30 is independently a linear C10-40 alkyl group;
each X1 is independently a single bond, -O-, -C(=O)-, -
C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
ring A1 is an optionally substituted C3-14 hydrocarbon
130
ring; and
n is an integer of 1 - 4, or
(2) optionally substituted C1-10 alkane having one or more
linear C10-40 alkyl groups via a group selected from the group
consisting of -O-, -C(=O)-, -C(=O)O-, -OC(=5 O)-, -C(=O)NH- and -
NHC(=O)-.
6. The precipitation promoter according to any one of claims 1
to 4, which is (1) a compound represented by the formula (I):
10
wherein
each R1 is independently a linear C10-40 alkyl group;
each X1 is independently a single bond, -O-, -C(=O)-, -
C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
ring A1 and ring B1 15 are each independently an optionally
substituted C3-14 hydrocarbon ring;
each X2 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q-, -(CH2)p-OC(=O)-
(CH2)q-, -(CH2)p-C(=O)NH-(CH2)q- or -(CH2)p-NHC(=O)-(CH2)q- (p and
20 q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-, -(CH2)r-
C(=O)O-, -(CH2)r-OC(=O)-, -(CH2)r-C(=O)NH- or -(CH2)r-NHC(=O)- (r
is an integer of 0 - 3);
R2 is a hydrogen atom, an optionally substituted C3-8
25 cycloalkyl group, an optionally substituted C1-6 alkyl group, an
optionally substituted C2-6 alkenyl group, an optionally
substituted C2-6 alkynyl group, an optionally substituted C6-14
aryl group, an optionally substituted monocyclic heterocyclic
group or a tri(C1-6 alkyl)silyl group;
30 n and m are each independently an integer of 1 - 4;
m’ is 0 or 1 when m is 1, and 1 when m is 2, 3 or 4, or
(2) optionally substituted C1-10 alkane having one or more
linear C10-40 alkyl groups via a group selected from the group
131
consisting of -O-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH- and -
NHC(=O)-.
7. The precipitation promoter according to claim 6, wherein R2
is a hydrogen atom, an optionally substituted 5 C3-8 cycloalkyl
group, an optionally substituted C1-6 alkyl group, an optionally
substituted C2-6 alkenyl group, an optionally substituted C2-6
alkynyl group, an optionally substituted C6-14 aryl group or an
optionally substituted monocyclic heterocyclic group.
10
8. The precipitation promoter according to any one of claims 1
to 4, which is a compound represented by the formula (II):
wherein
each R1 15 is independently a linear C10-40 alkyl group;
each X1 is independently a single bond, -O-, -C(=O)-, -
C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
ring A2 and ring B1 are each independently an optionally
substituted C3-14 hydrocarbon ring;
each X2 20 is independently -(CH2)p-, -(CH2)p-O-(CH2)q-, -
(CH2)p-C(=O)-(CH2)q-, -(CH2)p-C(=O)O-(CH2)q-, -(CH2)p-OC(=O)-
(CH2)q-, -(CH2)p-C(=O)NH-(CH2)q- or -(CH2)p-NHC(=O)-(CH2)q- (p and
q are each independently an integer of 0 - 3);
X3 is a single bond, -(CH2)r-O-, -(CH2)r-C(=O)-, -(CH2)r-
25 C(=O)O-, -(CH2)r-OC(=O)-, -(CH2)r-C(=O)NH- or -(CH2)r-NHC(=O)- (r
is an integer of 0 - 3);
R2 is a hydrogen atom, an optionally substituted C3-8
cycloalkyl group, an optionally substituted C1-6 alkyl group, an
optionally substituted C2-6 alkenyl group, an optionally
30 substituted C2-6 alkynyl group, an optionally substituted C6-14
aryl group, an optionally substituted monocyclic heterocyclic
132
group or a tri(C1-6 alkyl)silyl group;
m is an integer of 1 to 4; and
m’ is 0 or 1 when m is 1, and 1 when m is 2, 3 or 4.
9. The precipitation promoter according to claim 8, wherein R2 5
is a hydrogen atom, an optionally substituted C3-8 cycloalkyl
group, an optionally substituted C1-6 alkyl group, an optionally
substituted C2-6 alkenyl group, an optionally substituted C2-6
alkynyl group, an optionally substituted C6-14 aryl group or an
10 optionally substituted monocyclic heterocycle.
10. The precipitation promoter according to any one of claims 5
to 9, wherein the C3-14 hydrocarbon ring is selected from a
benzene ring and a cyclohexane ring.
15
11. The precipitation promoter according to any one of claims 1
to 10, wherein the organic group is a group represented by the
formula (III):
20
wherein
** shows a bonding position to a group to be protected;
L is a single bond, or a group represented by the formula
(a1) or (a1’):
25
wherein
* shows the bonding position to Y;
** is as defined above;
R8 and R9 are each independently a C1-22 hydrocarbon group;
30 L1 is a divalent C1-22 hydrocarbon group; and
133
L2 is a single bond, or a group represented by **C(=O)N(R2’)-R1’-
N(R3)*** wherein ** shows the bonding position to L1, *** shows
the bonding position to C=O, R1’ is a C1-22 alkylene group, and
R2’ and R3 are each independently a hydrogen atom or a C1-22
alkyl group, or R2’ and R3 are optionally joined 5 to form a ring,
Y is a single bond, an oxygen atom or NR wherein R is a
hydrogen atom, an alkyl group or an aralkyl group, and
Z is a group represented by the formula (a2), the formula (a2’)
or the formula (a2”):
10
wherein
* shows a bonding position;
R4 is a hydrogen atom, or when Rb is a group represented by the
following formula (a3), optionally joined with R6 of ring C to
15 show a single bond or -O- and to form a fused ring together
with ring A or ring B and ring C;
Q in the number of k are each independently -O-, -C(=O)-, -
C(=O)O-, -OC(=O)-, -C(=O)NH- or -NHC(=O)-;
R5 in the number of k are each independently a hydrocarbon
20 group bonded via a single bond or a linker to an aliphatic
hydrocarbon group having not less than 10 carbon atoms;
k is an integer of 1 to 4;
ring A and ring B are each independently optionally further
have, in addition to QR5 in the number of k, substituent(s)
25 selected from the group consisting of a halogen atom, a C1-6
alkyl group optionally substituted by a halogen atom, and a C1-6
alkoxy group optionally substituted by a halogen atom;
Ra is a hydrogen atom, or a phenyl group optionally substituted
by a halogen atom; and
30 Rb is a hydrogen atom, or a group represented by the formula
134
(a3):
wherein * shows a bonding position;
j is an integer of 0 to 4;
Q in the number of j are each independently 5 as defined above;
R7 in the number of j are each independently a hydrocarbon
group bonded via a single bond or a linker to an aliphatic
hydrocarbon group having not less than 10 carbon atoms;
R6 is a hydrogen atom, or optionally joined with R4 of ring A
10 or ring B to show a single bond or -O- and to form a fused ring
together with ring A or ring B and ring C; and
ring C optionally further has, in addition to OR7 in the number
of j, substituent(s) selected from the group consisting of a
halogen atom, a C1-6 alkyl group optionally substituted by a
15 halogen atom, and a C1-6 alkoxy group optionally substituted by
a halogen atom, or
Ra and Rb are joined to form an oxo group.
12. The precipitation promoter according to claim 11, wherein Z
20 is a group represented by the formula (a2) or the formula (a2’),
the fused ring is a fluorene ring or a xanthene ring, R5 in the
number of k are each independently a hydrocarbon group bonded
via a single bond or a linker to a linear aliphatic hydrocarbon
group having not less than 10 carbon atoms, R7 in the number of
25 j are each independently a hydrocarbon group bonded via a
single bond or a linker to a linear aliphatic hydrocarbon group
having not less than 10 carbon atoms, and Ra is a hydrogen atom,
or Ra and Rb are joined to form an oxo group.
30 13. The precipitation promoter according to claim 11 or 12,
wherein L is a group represented by the formula (a1), L1 is a
divalent C1-22 hydrocarbon group, and L2 is a single bond.
135
14. The precipitation promoter according to claim 11 or 12,
wherein L is a group represented by the formula (a1’), L1 is a
phenylene group, and L2 is a single bond.
5
15. The precipitation promoter according to any one of claims
11 to 14, wherein Y is an oxygen atom.
16. The precipitation promoter according to any one of claims
10 11 to 15, wherein Z is a group represented by the formula (a2),
and R4 is a hydrogen atom.
17. The precipitation promoter according to any one of claims
11 to 16, wherein Z is a group represented by the formula (a2),
15 and Ra and Rb are each a hydrogen atom.
18. The precipitation promoter according to claim 11 or 12,
wherein L and Y are each a single bond, Z is a group
represented by the formula (a2), R4 is a hydrogen atom, and Ra
20 and Rb are joined to form an oxo group.
19. The precipitation promoter according to any one of claims 1
to 18, wherein the organic compound protected by the organic
group is nucleoside, nucleotide or oligonucleotide optionally
25 further protected by a protecting group used in nucleic acid
synthesis, or amino acid or peptide optionally further
protected by a protecting group used in peptide synthesis.
20. The precipitation promoter according to any one of claims 1
30 to 18, wherein the organic compound protected by the organic
group is nucleoside, nucleotide or oligonucleotide optionally
further protected by a protecting group used in nucleic acid
synthesis.
35 21. The precipitation promoter according to any one of claims 1
136
to 18, wherein the organic compound protected by the organic
group is nucleoside or oligonucleotide wherein at least one
group selected from an amino group and an imino group of a
nucleic acid base, 2’- and 3’-hydroxy groups of a ribose
residue, and 3’-hydroxy group of a deoxyribose 5 residue is
protected by the organic group, and other group is optionally
further protected by a protecting group used in nucleic acid
synthesis.
10 22. The precipitation promoter according to any one of claims 1
to 21, wherein the solvent comprises a polar solvent.
23. The precipitation promoter according to claim 22, wherein
the solvent comprising a polar solvent is a mixed solvent of a
15 polar solvent and a nonpolar solvent.
24. The precipitation promoter according to claim 22 or 23,
wherein the polar solvent is acetonitrile.
20 25. The precipitation promoter according to any one of claims 1
to 24, which is used at not less than 0.1 molar equivalent
relative to the organic compound protected by the organic group
to precipitate the organic compound protected by the organic
group.
25
26. A precipitation mixture comprising the precipitation
promoter according to any one of claims 1 to 25, and an organic
compound protected by an organic group having one or more
aliphatic hydrocarbon groups having not less than 10 carbon
30 atoms.
27. The precipitation mixture according to claim 26, wherein
the aliphatic hydrocarbon group having not less than 10 carbon
atoms of the organic group is linear.
35
137
28. A method of precipitating an organic compound protected by
an organic group having one or more aliphatic hydrocarbon
groups having not less than 10 carbon atoms in a solvent, by
using the precipitation promoter according to any one of claims
5 1 to 25.
29. The method according to claim 28, wherein the solvent
comprises a polar solvent.
10 30. The method according to claim 29, wherein the polar solvent
is acetonitrile.
31. The method according to any one of claims 28 to 30, wherein
the aliphatic hydrocarbon group having not less than 10 carbon
15 atoms of the organic group is linear
32. A production method of an oligonucleotide, comprising a
step of adding a polar solvent to a reaction solution
comprising an oligonucleotide wherein at least one group is
20 protected by an organic group having one or more aliphatic
hydrocarbon groups having not less than 10 carbon atoms, and
other group is optionally further protected by a protecting
group used in nucleic acid synthesis, and the precipitation
promoter according to any one of claims 1 to 25 in a nonpolar
25 solvent, and separating a precipitate mixture comprising the
oligonucleotide and the precipitation promoter from the
reaction solution.
33. The production method according to claim 32, wherein the
30 polar solvent is acetonitrile.
34. The production method according to claim 32 or 33, wherein
the aliphatic hydrocarbon group having not less than 10 carbon
atoms of the organic group is linear.
35
138
35. The production method according to any one of claims 32 to
34, which is performed by a phosphoramidite method.
36. A production method of an oligonucleotide, which includes
one repeat of production cycle comprising the 5 following steps
(1) - (3), or plural repeats thereof by a phosphoramidite
method, which comprises the following step (4) in the first
cycle, the following step (5) in each cycle, and the following
step (6) in each cycle except the final cycle:
10 (1) a step of obtaining a reaction solution comprising a free-
5’-hydroxy-group form by adding an acid to a reaction solution
comprising a nucleoside or oligonucleotide wherein at least one
group selected from an amino group and an imino group of a
nucleic acid base, 2’- and 3’-hydroxy groups of a ribose
15 residue, and 3’-hydroxy group of a deoxyribose residue is
protected by an organic group having one or more aliphatic
hydrocarbon groups having not less than 10 carbon atoms, a 5’-
hydroxy group is protected by a temporary protecting group
removable under acidic conditions, and other group is
20 optionally further protected by a protecting group used in
nucleic acid synthesis in a nonpolar solvent, to deprotect the
temporary protecting group of the 5’-hydroxy group, and
neutralizing same with a base;
(2) a step of obtaining a reaction solution comprising a
25 phosphite triester form, by adding nucleoside or
oligonucleotide wherein a 3’-hydroxy group is phosphoramidited,
a 5’-hydroxy group is protected by a temporary protecting group
removable under acidic conditions, and other group is
optionally further protected by a protecting group used in
30 nucleic acid synthesis to the reaction solution comprising the
free-5’-hydroxy-group form in a nonpolar solvent;
(3) a step of obtaining a reaction solution comprising an
oligonucleotide wherein at least one group selected from an
amino group and an imino group of a nucleic acid base, 2’- and
35 3’-hydroxy groups of a ribose residue, and 3’-hydroxy group of
139
a deoxyribose residue is protected by an organic group having
one or more aliphatic hydrocarbon groups having not less than
10 carbon atoms, a 5’-hydroxy group is protected by a temporary
protecting group removable under acidic conditions, and other
group is optionally further protected by 5 a protecting group
used in nucleic acid synthesis, by adding an oxidizing agent or
a sulfurizing agent to the reaction solution comprising the
phosphite triester form in a nonpolar solvent;
(4) a step of adding the precipitation promoter according to
10 any one of claims 1 to 25 to the reaction solution at any of
before step (1), between steps (1) and (2), between steps (2)
and (3) and after step (3);
(5) a step of separating a precipitation mixture comprising the
free-5’-hydroxy-group form, the phosphite triester form or the
15 oligonucleotide, and the precipitation promoter from the
reaction solution by adding a polar solvent to the reaction
solution comprising the precipitation promoter at after step
(4), and any of between steps (1) and (2), between steps (2)
and (3) and after step (3);
20 (6) a step of adding a nonpolar solvent to the precipitation
mixture obtained in step (5) to give a reaction solution.
37. The production method according to claim 36, wherein the
polar solvent is acetonitrile.
25
38. The production method according to claim 36 or 37, wherein
the aliphatic hydrocarbon group having not less than 10 carbon
atoms of the organic group is linear.
30 39. The method according to any one of claims 36 to 38, wherein
the steps (5) and (6) are performed after step (3).
40. The method according to any one of claims 36 to 39, further
comprising the following step (7):
35 (7) a step of removing all protecting groups of the
140
oligonucleotide and isolating the oligonucleotide.
41. The method according to any one of claims 36 to 40, wherein
the non-polar solvent is a solvent selected from the group
consisting of a halogenated solvent, an aromatic 5 solvent, an
ester solvent, an aliphatic solvent, and a combination thereof.