Acetal-type releasable polyoxyethylene derivative containing amino group, method for producing the same, and acetal-type releasable polyoxyethylene conjugate containing amino group

Abstract

To provide a polyoxyethylene derivative having an amino group and an acetal structure, which converts a functional biomolecule into a prodrug and gradually releases the functional biomolecule under physiological conditions, and a method for producing the same.SOLUTION: There is provided an acetal-type releasable polyoxyethylene derivative containing an amino group, which is, for example, represented by a formula (1). (B1 represents a hydrogen atom or -C(R1)(R2)OC(O)E1, R1 and R2 each independently represent a hydrocarbon group having 1 to 10 carbon atoms or H, E1 represents a leaving group, P1 represents a polyoxyethylene derivative moiety, w represents an integer of 1 to 8, and m represents 0 or 1).SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to an acetal-type releasable polyoxyethylene derivative containing an amino group, a method for producing the same, and an acetal-type releasable polyoxyethylene conjugate containing an amino group, which is a conjugate with a biofunctional molecule. [Background technology]

[0002] When drugs using biofunctional molecules such as hormones, cytokines, and enzymes are administered to the body, they are usually quickly excreted from the body by glomerular filtration in the kidneys or uptake by macrophages in the liver and spleen. As a result, their half-life in the blood is short, and it is often difficult to obtain sufficient pharmacological effects. To solve this problem, attempts have been made to chemically modify biofunctional molecules with water-soluble polymers such as polyoxyethylene (PEG). As a result, it is possible to extend the half-life in the blood of biofunctional molecules by increasing the molecular weight and forming a hydration layer. It is also well known that these modifications can reduce the toxicity and antigenicity of biofunctional molecules and improve their aggregation properties.

[0003] On the other hand, it is known that biofunctional molecules chemically modified with water-soluble polymers such as PEG can have undesirable effects such as a decrease in the inherent pharmacological action of the biofunctional molecules and changes in their dynamics in the body and within cells due to the formation of a hydration layer by the water-soluble polymer and the steric shielding effect of the active site, which reduces the interaction with the target endogenous molecules and receptors.

[0004] As an approach to the above-mentioned problems, a method is used in which a water-soluble polymer is chemically modified to a biofunctional molecule via a temporary bond, and the temporary bond is cleaved in vivo to release the unmodified biofunctional molecule, i.e., a prodrug method.

[0005] In recent years, the techniques described in Patent Documents 1 and 2 have been known as techniques that can suppress the decrease in the inherent pharmacological action of biofunctional molecules and changes in the dynamics in the body and within cells of biofunctional molecules modified with water-soluble polymers by using a prodrug method. These Patent Documents 1 and 2 report that PEGylated hGH and PEGylated IL-2, which are biofunctional molecules modified with PEG via a linker that decomposes non-enzyme-dependently at an appropriate rate under physiological conditions, i.e., neutral conditions, can improve the decrease in pharmacological action caused by PEGylation by releasing the biofunctional molecule with the decomposition of the linker while extending the half-life in the blood of the biofunctional molecule. For this reason, a prodrug technique that extends the half-life of the biofunctional molecule in the blood and releases the biofunctional molecule at an appropriate rate under physiological conditions in an enzyme-independent manner to express a pharmacological action is important.

[0006] Patent Document 3 reports a compound that non-enzyme-dependently cleaves a temporary bond in a biofunctional molecule that has been chemically modified with PEG, thereby releasing the biofunctional molecule that has not been chemically modified.

[0007] Specifically, it has been reported that the hydrolysis of a linker having an acetal structure that is hydrolyzed under acidic conditions triggers cleavage of the temporary carbamate bond through 1,4- or 1,6-benzyl elimination, releasing an unmodified biofunctional molecule. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] JP 2018-150311 A [Patent Document 2] Patent Publication No. 2022-000043 [Patent Document 3] JP 2018-172648 A Summary of the Invention [Problem to be solved by the invention]

[0009] Incidentally, in the examples of Patent Document 3, structures capable of releasing biofunctional molecules in an enzyme-independent manner and under acidic conditions are exemplified, but no prodrug technology is described that releases biofunctional molecules at an appropriate rate in an enzyme-independent manner under physiological conditions, i.e., under neutral conditions around pH 7.4, and exerts a pharmacological action.

[0010] In view of the above problems, the object of the present invention is to provide a polyoxyethylene derivative having an amino group and an acetal structure, which is characterized by converting a biofunctional molecule into a prodrug and gradually releasing the biofunctional molecule under physiological conditions, a stable production method for the same, and an acetal-type releasable polyoxyethylene conjugate containing an amino group. [Means for solving the problem]

[0011] As a result of extensive research, the present inventors have discovered a polyoxyethylene derivative having an amino group and an acetal structure, which can convert a biofunctional molecule into a prodrug and gradually hydrolyze the acetal under physiological conditions to induce benzyl elimination, thereby releasing the biofunctional molecule, as well as a stable method for producing the same, and an acetal-type releasable polyoxyethylene conjugate containing an amino group.

[0012] That is, the present invention relates to the following [1] to

[10] . [1] An acetal-type releasable polyoxyethylene derivative containing an amino group, characterized in that it is represented by the following formula (1), formula (2), formula (3) or formula (4): [ka] [ka] [ka] [ka] (In formulas (1), (2), (3) and (4), B 1 is a hydrogen atom or -C(R 1 )(R 2 )OC(O)E 1 and E 1 is a leaving group, P 1 is a polyoxyethylene derivative moiety, w is an integer from 1 to 8; R 1 , R 2 , R 3 , R 4 , R 5 and R 12 each independently represents a hydrocarbon group having 1 to 10 carbon atoms or a hydrogen atom, R 6 represents an optionally substituted hydrocarbon group having 1 to 24 carbon atoms or a hydrogen atom, R 7 , R 8 , R 9 , R 10 and R 11 each independently represents an electron-withdrawing substituent, an electron-donating substituent, or a hydrogen atom; R 13 represents an optionally substituted hydrocarbon group having 1 to 10 carbon atoms or a hydrogen atom, R 14 represents an optionally substituted hydrocarbon group having 1 to 10 carbon atoms, R 13 is R 14 and m is 0 or 1.)

[0013] [2] m is 0 and R 1 and R 2 is a hydrogen atom, and R 3 , R 4 , R 5 and R 12and each independently represents a hydrogen atom or a methyl group.

[0014] [3] m is 0 and R 1 and R 2 is a hydrogen atom, and R 3 , R 4 , R 5 and R 12 are each independently a hydrogen atom or a methyl group, and R 13 and R 14 and are combined, R 13 is a hydrocarbon group represented by the following formula (5), R 14 The amino group-containing acetal-type releasable polyoxyethylene derivative of [1] or [2] is characterized in that the hydrocarbon group is represented by the following formula (6): [ka] (In formula (5), R 15 and R 16 each independently represents an optionally substituted hydrocarbon group having 1 to 4 carbon atoms or a hydrogen atom, q1 is an integer from 1 to 4, (*1) indicates the point of attachment to N, (*2) is R 14 ) [ka] (In formula (6), R 17 and R 18 each independently represents an optionally substituted hydrocarbon group having 1 to 4 carbon atoms or a hydrogen atom, R 19 is CR 20 or a nitrogen atom, R 20represents an optionally substituted hydrocarbon group having 1 to 4 carbon atoms or a hydrogen atom, q2 is an integer from 0 to 3; Z 1 is R 19 Carbon or nitrogen atoms and P 1 is a single bond or a divalent spacer connecting (*3) indicates the point of attachment to N, (*4) is R 13 represents the point of attachment to (*5) is P 1 )

[0015] [4] m is 0, R 1 and R 2 is a hydrogen atom, R 3 , R 4 , R 5 and R 12 are each independently a hydrogen atom or a methyl group, R 13 and R 14 is bonded, R 15 and R 16 is a hydrogen atom, q1 is 2, R 17 and R 18 is a hydrogen atom, R 19 is a methine group or a nitrogen atom, An acetal-type releasable polyoxyethylene derivative having an amino group represented by [3], wherein q2 is 2.

[0016] [5] A method for producing an acetal-type releasable polyoxyethylene derivative having an amino group represented by [1], [2], [3] or [4], comprising the steps of: a coupling step of coupling a polyoxyethylene derivative with a benzaldehyde derivative or a phenyl ketone derivative to obtain a coupling product represented by the following formula (7) or (8); After the coupling step, the coupling product represented by the following formula (7) or (8) is reacted under acidic conditions with a hydroxymethyl group at the 2-position and a substituent (-CH=CB 1 )C(R 1 )(R 2 )-OH(B 1 , m, R 1 , R 2 (wherein m is as described above) with a phenol to obtain an acetal structure; and After the acetalization step, a leaving group structure (-OC(O)E) is added to the terminal of the substituent at the 4-position or 6-position. 1 )(E 1 is a leaving group) 1. A method for producing an acetal-type releasable polyoxyethylene derivative having an amino group, comprising: [ka] [ka] (In formula (7) and formula (8), P 1 ,w,R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 13 and R 14 is as mentioned above.)

[0017] [6] A method for producing an acetal-type releasable polyoxyethylene derivative having an amino group according to [5], characterized in that a dialkylation step is provided between the coupling step and the acetalization step, in which the coupling product of formula (7) or formula (8) is reacted with a monohydric alcohol under acidic conditions to obtain a dialkyl acetal structure represented by formula (9) or formula (10): [ka] [ka] (In formulas (9) and (10), R 21 and R 22 are each independently a hydrocarbon group having 1 to 10 carbon atoms, P 1 ,w,R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 13 and R 14 is as mentioned above.)

[0018] [7] a deprotection step of an amino group protected by a protecting group in the coupling product between the acetalization step and the leaving group structure introduction step; and 6. A method for producing an acetal-type releasable polyoxyethylene derivative having an amino group according to claim 5, further comprising the step of introducing a functional group capable of reacting with a biofunctional molecule to the amino group deprotected after the deprotection step.

[0019] [8] a deprotection step of an amino group protected by a protecting group in the dialkyl acetal structure between the acetalization step and the leaving group structure introduction step; and A method for producing an acetal-type releasable polyoxyethylene derivative having an amino group according to [6], characterized in that it comprises a step of introducing a functional group capable of reacting with a biofunctional molecule into the amino group deprotected after the deprotection step.

[0020] [9] An acetal-type releasable polyoxyethylene conjugate containing an amino group, which is represented by the following formula (11), formula (12), formula (13) or formula (14) and is characterized in that it is cleaved under physiological conditions. [ka] [ka] [ka] [ka] (In formula (11), formula (12), formula (13) and formula (14), B 2 is a hydrogen atom or -C(R 1 )(R 2 )OC(O)NHD 1 and D 1 is a residue obtained by removing an amino group constituting a carbamate bond from an amino group contained in a biofunctional molecule, P 2 is a polyoxyethylene derivative portion or a conjugate of a polyoxyethylene derivative portion and a biofunctional molecule, w is an integer from 1 to 8; R 1 , R 2 , R 3 , R 4 , R 5 and R 12 each independently represents a hydrocarbon group having 1 to 10 carbon atoms or a hydrogen atom, R 6 represents an optionally substituted hydrocarbon group having 1 to 24 carbon atoms or a hydrogen atom, R 7 , R 8 , R 9 , R 10 and R 11 are each independently an electron-withdrawing substituent, an electron-donating substituent, or a hydrogen atom; R 13 represents an optionally substituted hydrocarbon group having 1 to 10 carbon atoms or a hydrogen atom, R 14represents an optionally substituted hydrocarbon group having 1 to 10 carbon atoms, R 13 is R 14 and m is 0 or 1.)

[0021]

[10] A method for producing an acetal-type releasable polyoxyethylene conjugate according to [9], comprising a coupling step of reacting an acetal-type releasable polyoxyethylene derivative according to any one of [1] to [4] with a biofunctional molecule in a neutral or basic buffer solution which may contain a water-soluble organic solvent; A purification step under neutral or basic conditions after the coupling step. A method for producing an acetal-type releasable polyoxyethylene conjugate containing an amino group, comprising: Effect of the Invention

[0022] The acetal-type releasable polyoxyethylene derivative containing an amino group of the present invention converts a biofunctional molecule into a prodrug, and gradually hydrolyzes the acetal under physiological conditions to induce benzyl elimination, gradually releasing the biofunctional molecule, thereby improving the pharmacological action of the biofunctional molecule modified with the polyoxyethylene derivative.

[0023] In the examples of Patent Document 3, a structure capable of releasing a biofunctional molecule in an enzyme-independent manner and under acidic conditions is exemplified, but a specific structure capable of releasing a biofunctional molecule by benzyl elimination triggered by acetal hydrolysis under physiological conditions, i.e., neutral conditions at about pH 7.4, is not described.

[0024] Here, the acetal structure in the compound of Patent Document 3 is generally used as a protecting group for diol or carbonyl groups, and it is known that the deprotection is performed under acidic conditions, and it is stable under neutral or basic conditions. Therefore, the compound of Patent Document 3 does not suggest the creation of a compound that is hydrolyzed at an appropriate rate under physiological conditions, i.e., neutral conditions, to release a biofunctional molecule. [Brief description of the drawings]

[0025] [Figure 1] The results are from a decomposition test of the compounds of formulae (47), (48), (49) and (50) in phosphate buffered saline at pH 7.4 at 37°C. [Diagram 2] The results are from a decomposition test of the compounds of formulae (63), (64), (65) and (66) in phosphate buffered saline at pH 7.4 at 37°C. [Diagram 3] The amino acid sequence of IL-2. [Figure 4] These are the results of a CTLL-2 cell proliferation test using (RLS) PEG(20k)-IL2 and (NRL) PEG(20k)-IL2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0026] The present invention will be described in detail below. The present invention is characterized in that, under physiological conditions, the acetal undergoes hydrolysis, followed by benzyl elimination, and the biofunctional molecule is gradually released. In this specification, the physiological conditions are pH 6.0 to 8.0.

[0027] The release rate of a biofunctional molecule under physiological conditions can be evaluated as the half-life of an acetal in a buffer solution of pH 7.4. The half-life of an acetal in a buffer solution of pH 7.4 is preferably 1 to 840 hours. Here, the "half-life" refers to the time required for an acetal to be hydrolyzed and 1 / 2 equivalent to become an aldehyde.

[0028] The above-mentioned acetal half-life under physiological conditions could not be achieved with a compound in which a polyoxyethylene derivative is introduced via an ether bond to the 3-position of the phenyl group bonded to the acetal, as exemplified in the examples of Patent Document 3. In contrast, the present inventors have found that a compound in which a polyoxyethylene derivative is introduced via an amino group to the 2- or 4-position of the phenyl group bonded to the acetal can achieve the above-mentioned acetal half-life when applied under physiological conditions.

[0029] That is, the present invention relates to an acetal-type releasable polyoxyethylene derivative containing an amino group, which is represented by the following formula (1), formula (2), formula (3) or formula (4) and is characterized by being cleaved under physiological conditions. [ka] [ka] [ka] [ka]

[0030] In formulas (1), (2), (3) and (4), B 1 is a hydrogen atom or -C(R 1 )(R 2 )OC(O)E 1 and preferably a hydrogen atom.

[0031] E 1is a leaving group. Preferred examples of the leaving group include succinimidyloxy group, phthalimidyloxy group, 4-nitrophenoxy group, 1-imidazolyl group, pentafluorophenoxy group, benzotriazol-1-yloxy group, and 7-azabenzotriazol-1-yloxy group, more preferably succinimidyloxy group, 4-nitrophenoxy group, 1-imidazolyl group, and pentafluorophenoxy group, and even more preferably succinimidyloxy group or 4-nitrophenoxy group.

[0032] P 1 is a polyoxyethylene derivative portion, and the polyoxyethylene derivative portion includes both polyoxyethylene having a molecular weight distribution obtained by polymerization of ethylene oxide, and monodisperse polyoxyethylene obtained by coupling oligooxyethylene having a single molecular weight by a coupling reaction. 1 The polyoxyethylene derivative portion may have a hydrocarbon group having 1 to 24 carbon atoms, an amino group protected by a protecting group, or a functional group capable of reacting with a biofunctional molecule. and w is an integer from 1 to 8.

[0033] P 1 Preferred examples of the residues are as follows, depending on the number of w: When w=1, P 1 Preferred examples of the above are residues represented by the following formula (p1) or (p2): [ka] (X 1 is a hydrocarbon group having 1 to 24 carbon atoms, an amino group protected by a protecting group, or a functional group capable of reacting with a biofunctional molecule, Z 2 and Z 3 is a divalent spacer or a single bond, n and l each independently represent 3 to 2,000; s is 0 or 1, t is 2 or 3, and v is 0 or 2. When w=2, P 1Preferred examples of the above are residues represented by the following formula (p3) or (p4). [ka] (X 1 , Z 2 , Z 3 , n, l, s and t are as defined above.) When w=3, P 1 A preferred example is a residue represented by the following formula (p5): [ka] (Z 3 and n has the same meaning as above.) When w=4, P 1 Preferred examples are residues represented by the following formulae (p6), (p7), and (p8). [ka] (X 1 , Z 2 , Z 3 , n, l, s and t are as defined above.) When w=5, P 1 A preferred example is a residue represented by the following formula (p9). [ka] (Z 3 and n has the same meaning as above.) When w=6, P 1 A preferred example is a residue represented by the following formula (p10): [ka] (Z 3 and n has the same meaning as above.) When w=8, P 1 Preferred examples are residues represented by the following formulae (p11), (p12), and (p13). [ka] (Z 2 and n has the same meaning as above.) (*6) is L 1 Indicates the connection point with

[0034] In formulas (p1) to (p13), n and l are -(CH 2 CH 2 Each of these independently represents the number of moles of oxyethylene groups added, represented by -(CH 2 CH 2 O) n The number of moles of oxyethylene groups added can be calculated by subtracting the molecular weight derived from molecules other than polyoxyethylene chains, represented by -, and dividing the result by 44, which is derived from the oxyethylene groups.

[0035] In formulas (p1), (p2), (p4) and (p8), Z 2 is a polyoxyethylene group and X 1 In formulae (p1) to (p13), Z 3 is a polyoxyethylene group and R 14 is a divalent spacer or single bond connecting 2 and Z 3 The divalent spacer is not particularly limited as long as it is more stable than an acetal structure, but is preferably an ether bond, an ester bond, a carbonate bond, a urethane bond, an amide bond, a secondary amino group, or an alkylene group containing these, or a single bond, and the alkylene group has 1 to 24 carbon atoms. Examples of the divalent spacer include spacers (z1) to (z8) described in group (I).

[0036] Group (I) [ka] In the formula, q3 and q4 are each independently an integer of 1 to 12. For example, when it is desired to bond the terminal active carbonate group in a hydrophobic environment such as the inside of a protein, it is preferable that q3 and q4 are large, and when it is desired to bond in a hydrophilic environment, it is preferable that q3 and q4 are small. However, Z 2 and Z 3 Each of them is an ether bond, an ester bond, a carbonate bond, a urethane bond, an amide bond, a secondary amino group, or an alkylene group containing these, and when a plurality of the same structural units are bonded, the number of said structural units is 2 or less.

[0037] Next, P 1 The polyoxyethylene derivative portion of the formula (p1), (p2), (p4), and (p8) may have a hydrocarbon group having 1 to 24 carbon atoms, an amino group protected by a protecting group, or a functional group capable of reacting with a biofunctional molecule. These are represented by X in the formulas (p1), (p2), (p4), and (p8). 1 This also applies to the following.

[0038] Specific examples of the hydrocarbon group include a methyl group, an ethyl group, a propyl group, an isopropyl group, an allyl group, a butyl group, a t-butyl group, a pentyl group, an isopentyl group, a hexyl group, a heptyl group, a 2-ethylhexyl group, an octyl group, a nonyl group, a decyl group, an undecyl group, a dodecyl group, a tridecyl group, a tetradecyl group, a pentadecyl group, a hexadecyl group, a heptadecyl group, an octadecyl group, a nonadecyl group, an eicosyl group, a heneicosyl group, a docosyl group, a toicosyl group, a tetracosyl group, a phenyl group, a benzyl group, a cresyl group, a butylphenyl group, a dodecylphenyl group, and a trityl group. A hydrocarbon group having 1 to 10 carbon atoms is preferable, a methyl group or an ethyl group is more preferable, and a methyl group is even more preferable.

[0039] Here, a protecting group is a component that prevents or inhibits the reaction of a specific chemically reactive functional group in a molecule under certain reaction conditions. The protecting group varies depending on the type of chemically reactive functional group to be protected, the conditions used, and the presence of other functional groups or protecting groups in the molecule. Specific examples of protecting groups can be found in many general textbooks, for example, "Wuts, PGM; Greene, TW Protective Groups in Organic Synthesis, 4th ed.; Wiley-Interscience: New York, 2007". In the present invention, the amino group protected by a protecting group includes, for example, an amino group protected by an acyl-based protecting group or a carbamate-based protecting group, or an azide group, and specific examples of the acyl-based protecting group or the carbamate-based protecting group include a trifluoroacetyl group, a 9-fluorenylmethyloxycarbonyl group, and a 2-(trimethylsilyl)ethyloxycarbonyl group.

[0040] Furthermore, the functional group capable of reacting with a biofunctional molecule is a formyl group, an epoxy group, a maleimidyl group, a vinylsulfone group, an acryl group, a sulfonyloxy group, a carboxy group, a dithiopyridyl group, an α-haloacetyl group, an alkynyl group, an allyl group, a vinyl group, or an azide group.

[0041] More specifically, functional groups capable of reacting with an amino group of a biofunctional molecule to form a covalent bond include formyl, epoxy, maleimidyl, vinylsulfone, acryl, sulfonyloxy, and carboxy groups. Functional groups capable of reacting with a thiol group of a biofunctional molecule to form a covalent bond include formyl, epoxy, maleimidyl, vinylsulfone, acryl, sulfonyloxy, carboxy, dithiopyridyl, α-haloacetyl, alkynyl, allyl, and vinyl groups. Functional groups capable of reacting with an alkynyl group of a biofunctional molecule to form a covalent bond include an azide group. Functional groups capable of reacting with an azide group of a biofunctional molecule to form a covalent bond include an alkynyl group and a triple bond-containing functional group.

[0042] In a preferred embodiment of this aspect, the functional group capable of reacting with a biofunctional molecule is a group represented by group (II), group (III), group (IV) or group (V). 2 Represents the connection point with. Group (II): Functional groups capable of reacting with amino groups of biofunctional molecules to form covalent bonds Items (a), (b), (e) and (f) below. Group (III): Functional groups capable of reacting with a thiol group of a biofunctional molecule to form a covalent bond (a), (b), (c), (d), (e), (f) and (h) below. Group (IV): Functional groups capable of reacting with an alkynyl group of a biofunctional molecule to form a covalent bond Items (c), (d) and (g) below. Group (V): Functional groups capable of reacting with an azide group of a biofunctional molecule to form a covalent bond (i) and (j) below. [ka] In the formula, Y 1 and Y 3 are each independently a hydrogen atom or a hydrocarbon group having 1 to 5 carbon atoms, and specific examples of the hydrocarbon group include a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, a t-butyl group, and a pentyl group. 2 is a halogen atom selected from a chlorine atom, a bromine atom, and an iodine atom.

[0043] In formula (p2), formula (p4) and formula (p8), s is 0 or 1, t is 2 or 3, and v is 0 or 2. Preferred embodiments are residues represented by the following formulae (p14) to (p19).

[0044] [ka] In formulas (p14) to (p19), X 1 , Z 2 , Z3 , n, l and (*6) are as defined above.

[0045] In formulas (1) to (4), R 1 , R 2 , R 3 , R 4 , R 5 and R 12 are each independently a hydrocarbon group having 1 to 10 carbon atoms or a hydrogen atom. Specific examples of the hydrocarbon group include a methyl group, an ethyl group, a propyl group, an isopropyl group, a t-butyl group, a phenyl group, and a benzyl group, and a hydrogen atom or a methyl group is more preferable.

[0046] In formulas (1) to (4), R 6 is a hydrogen atom or a hydrocarbon group having 1 to 24 carbon atoms which may be substituted. Specific examples of the hydrocarbon group which may be substituted include a methyl group, a fluoromethyl group, a difluoromethyl group, a trifluoromethyl group, a bromomethyl group, a dibromomethyl group, a tribromomethyl group, a chloromethyl group, a dichloromethyl group, a trichloromethyl group, an ethyl group, a propyl group, a isopropyl group, a butyl group, a t-butyl group, a pentyl group, an isopentyl group, a hexyl group, a heptyl group, a 2-ethylhexyl group, an octyl group, a nonyl group, a decyl group, an undecyl group, a dodecyl group, a tridecyl group, a tetradecyl group, a pentadecyl group, a hexa ... Examples of such groups include a decyl group, a heptadecyl group, an octadecyl group, a nonadecyl group, an eicosyl group, a heneicosyl group, a docosyl group, a tricosyl group, a tetracosyl group, a phenyl group, a benzyl group, a cresyl group, a butylphenyl group, a dodecylphenyl group, and a trityl group, and preferred are a methyl group, a fluoromethyl group, a difluoromethyl group, a trifluoromethyl group, a bromomethyl group, a dibromomethyl group, a tribromomethyl group, a chloromethyl group, a dichloromethyl group, a trichloromethyl group, an ethyl group, or an optionally substituted phenyl group.

[0047] The optionally substituted phenyl group is represented by the following formula (15): [ka] In the formula, R 23 ~R 27 are each independently an electron-withdrawing substituent, an electron-donating substituent, or a hydrogen atom. Examples of the electron-withdrawing substituent include an acyl group having 2 to 5 carbon atoms, an alkoxycarbonyl group having 2 to 5 carbon atoms, a carbamoyl group having 2 to 5 carbon atoms, an acyloxy group having 2 to 5 carbon atoms, an acylamino group having 2 to 5 carbon atoms, an alkoxycarbonylamino group having 2 to 5 carbon atoms, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, an alkylsulfanyl group having 1 to 4 carbon atoms, an alkylsulfonyl group having 1 to 4 carbon atoms, an arylsulfonyl group having 6 to 10 carbon atoms, a nitro group, a trifluoromethyl group, and a cyano group. Preferred examples include an acetyl group, a methoxycarbonyl group, a methylcarbamoyl group, an acetoxy group, an acetamide group, a methoxycarbonylamino group, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a methylsulfanyl group, a phenylsulfonyl group, a nitro group, a trifluoromethyl group, and a cyano group.

[0048] The electron-donating substituent is an alkyl group having 1 to 4 carbon atoms, and preferred examples thereof include a methyl group, an ethyl group, a propyl group, an isopropyl group, and a t-butyl group. 23 , R 25 or R 27 Examples of the electron-donating substituent at the R position include an alkoxy group having 1 to 4 carbon atoms, an aryl group having 6 to 10 carbon atoms, and an aryloxy group having 6 to 10 carbon atoms. Preferred examples include a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a t-butoxy group, a phenyl group, and a phenoxy group. 24 or R 26 , electron-withdrawing, para and ortho positions, i.e., R 23 , R 25 or R 27 In the above, examples of the electron-donating substituent include an alkoxy group having 1 to 4 carbon atoms, an aryl group having 6 to 10 carbon atoms, and an aryloxy group having 6 to 10 carbon atoms. Preferred examples include a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a t-butoxy group, a phenyl group, and a phenoxy group. (*8) indicates the point of attachment to the carbon atom of the acetal structure.

[0049] In formulas (1) to (4), R 7 , R 8 , R 9 , R 10 and R 11 are each independently an electron-withdrawing substituent, an electron-donating substituent, or a hydrogen atom. Examples of the electron-withdrawing substituent include an acyl group having 2 to 5 carbon atoms, an alkoxycarbonyl group having 2 to 5 carbon atoms, a carbamoyl group having 2 to 5 carbon atoms, an acyloxy group having 2 to 5 carbon atoms, an acylamino group having 2 to 5 carbon atoms, an alkoxycarbonylamino group having 2 to 5 carbon atoms, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, an alkylsulfanyl group having 1 to 4 carbon atoms, an alkylsulfonyl group having 1 to 4 carbon atoms, an arylsulfonyl group having 6 to 10 carbon atoms, a nitro group, a trifluoromethyl group, and a cyano group. Preferred examples include an acetyl group, a methoxycarbonyl group, a methylcarbamoyl group, an acetoxy group, an acetamide group, a methoxycarbonylamino group, a fluorine atom, a chlorine atom, a bromine atom, an iodine atom, a methylsulfanyl group, a phenylsulfonyl group, a nitro group, a trifluoromethyl group, and a cyano group.

[0050] The electron-donating substituent is an alkyl group having 1 to 4 carbon atoms, and preferred examples thereof include a methyl group, an ethyl group, a propyl group, an isopropyl group, and a t-butyl group. 7 , R 10 or R 11 Examples of the electron-donating substituent at the R position include an alkoxy group having 1 to 4 carbon atoms, an aryl group having 6 to 10 carbon atoms, and an aryloxy group having 6 to 10 carbon atoms. Preferred examples include a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a t-butoxy group, a phenyl group, and a phenoxy group. 8 or R 9 , electron-withdrawing, para and ortho positions, i.e., R 7 , R 10 or R 11In the above, examples of the electron-donating substituent include an alkoxy group having 1 to 4 carbon atoms, an aryl group having 6 to 10 carbon atoms, and an aryloxy group having 6 to 10 carbon atoms. Preferred examples include a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a t-butoxy group, a phenyl group, and a phenoxy group.

[0051] In formulas (1) to (4), R 13 is an optionally substituted hydrocarbon group having 1 to 10 carbon atoms or a hydrogen atom, and R 14 is an optionally substituted hydrocarbon group having 1 to 10 carbon atoms, R 13 is R 14 may be bonded to

[0052] R 13 and R 14 is preferably in the following form depending on the presence or absence of a bond: (R 13 and R 14 (If not combined) R 13 is preferably a methyl group, ethyl group, propyl group, isopropyl group, butyl group, t-butyl group, pentyl group, isopentyl group, hexyl group, heptyl group, or a hydrogen atom, more preferably a methyl group, ethyl group, propyl group, isopropyl group, butyl group, t-butyl group, or a hydrogen atom, and even more preferably a methyl group, ethyl group, propyl group, isopropyl group, or a hydrogen atom.

[0053] R 14 represents an alkylene group which may contain an ether bond, an ester bond, a carbonate bond, a urethane bond, an amide bond, or a secondary amino group, and the alkylene group has 1 to 10 carbon atoms. Examples of the alkylene group include the divalent spacers (z1) to (z8) described in the above-mentioned group (I), and the sum of q3 and q4 in (z1) to (z8) is preferably 1 to 10.

[0054] A more preferred embodiment is R 13 is a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, a t-butyl group, or a hydrogen atom, and R 14is (z1), (z2), (z3), (z6), (z7) or (z8). R 13 and R 14 A further preferred embodiment when is not bound is 13 is a methyl group, an ethyl group, a propyl group, an isopropyl group, or a hydrogen atom, and R 14 is more preferably (z1), (z2) or (z7).

[0055] (R 13 and R 14 (If R 13 is expressed by the following formula (5) 14 is an optionally substituted hydrocarbon group having 1 to 10 carbon atoms connecting the [ka] R 15 and R 16 are each independently an optionally substituted hydrocarbon group having 1 to 4 carbon atoms or a hydrogen atom, q1 is an integer of 1 to 4, (*1) is the point of attachment to N, (*2) is R 14 R 15 and R 16 is more preferably a methyl group, an ethyl group or a hydrogen atom.

[0056] R 14 is expressed by the following equation (6): 1 is an optionally substituted hydrocarbon group having 1 to 10 carbon atoms connecting the [ka]

[0057] In formula (6), R 17 and R 18 are each independently an optionally substituted hydrocarbon group having 1 to 4 carbon atoms or a hydrogen atom, R 19 is CR 20 or a nitrogen atom, R 20represents an optionally substituted hydrocarbon group having 1 to 4 carbon atoms or a hydrogen atom, q2 represents an integer of 0 to 3, and Z 1 is CR 20 Carbon or nitrogen atoms and P 1 is a single bond or a divalent spacer that connects the two, (*3) represents the point of attachment to N, and (*4) represents R 13 (*5) represents the point of attachment to P 1 Represents the connection point with.

[0058] R 17 , R 18 and R 19 Preferred examples of the radical are a methyl group, an ethyl group, a propyl group, a butyl group, an isopropyl group, a t-butyl group, or a hydrogen atom, and a methyl group, an ethyl group, or a hydrogen atom are more preferred.

[0059] Z 1 is a single bond or a divalent spacer. The divalent spacer is not particularly limited as long as it is more stable than an acetal structure, but is preferably an ether bond, an ester bond, a carbonate bond, a urethane bond, an amide bond, a secondary amino group, or an alkylene group containing these, or a single bond. The alkylene group has 1 to 24 carbon atoms. Examples of the spacers include (z1) to (z8) described in the above-mentioned group (I), and the detailed explanation of (z1) to (z8) is the same as described above.

[0060] R 13 and R 14 A more preferred embodiment when is attached is R 13 Medium, R 15 and R 16 is a methyl group, an ethyl group, or a hydrogen atom, q1 is 1 to 4, and R 14 Medium, R 17 and R 18 is a methyl group, an ethyl group, or a hydrogen atom, and R 19 CR 20 or a nitrogen atom, R 20 is a methyl group, an ethyl group, or a hydrogen atom, q2 is 0 to 3, the sum of q1 and q2 is 2 to 5, and Z 1is a single bond, (z1), (z2), (z3), (z6), (z7) or (z8).

[0061] R 13 and R 14 A further preferred embodiment when is attached is R 13 Medium, R 15 and R 16 is a methyl group or a hydrogen atom, q1 is 1 to 4, and R 14 Medium, R 17 and R 18 is a methyl group or a hydrogen atom, and R 19 CR 20 or a nitrogen atom, R 20 is a methyl group or a hydrogen atom, q2 is 0 to 3, the sum of q1 and q2 is 3 to 5, and Z 1 is a single bond, (z1), (z2), (z3), (z6) or (z7).

[0062] In a more preferred embodiment of the present invention, in the formulas (1) to (4), m is 0 and R 1 and R 2 is a hydrogen atom, and R 3 , R 4 , R 5 and R 11 is a hydrogen atom or a methyl group having one carbon atom, and R 3 , R 4 and R 5 is a hydrogen atom, and R 11 More preferably, is a methyl group.

[0063] In a further preferred embodiment of the present invention, in the formulas (1) to (4), m is 0 and R 1 and R 2 is a hydrogen atom, and R 3 , R 4 , R 5 and R 12 are each independently a hydrogen atom or a methyl group, and L 1 R in 13 and R 14 is bonded, R 13 Ga-CH 2 -CH 2-, i.e., R 15 and R 16 is a hydrogen atom and q1 is 2, and R 14 Ga-CH 2 -CH 2 -CH< or -CH 2 -CH 2 -N<, i.e., R 17 and R 18 is a hydrogen atom, and R 19 is a methine group or a nitrogen atom, q2 is 2, and Z 1 It is more preferable that is a single bond or a methylene group.

[0064] Next, a method for producing the acetal-type releasable polyoxyethylene derivative containing an amino group according to the present invention will be described. The present invention provides a method for producing an acetal-type releasable polyoxyethylene derivative having an amino group, comprising the steps of: a coupling step of coupling a polyoxyethylene derivative with a benzaldehyde derivative or a phenyl ketone derivative to obtain a coupling product represented by the following formula (7) or (8); and after the coupling step, a coupling product represented by the formula (7) or (8) is converted under acidic conditions into a hydroxymethyl group having a hydroxymethyl group at the 2-position and a substituent (-CH=CB) at the 4-position or the 6-position. 1 )C(R 1 )(R 2 )-OH(B 1 , m, R 1 , R 2 (as described above) to obtain an acetal structure; and (ii) providing a leaving group structure (-OC(O)E at the end of the substituent at the 4-position or 6-position after the acetalization step. 1 )(E 1 is as described above), a leaving group structure introduction step is carried out.

[0065] [ka] [ka] (In the formulas (7) and (8), P 1 ,w,R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 13 and R 14 is as mentioned above.)

[0066] The coupling step of a polyoxyethylene derivative with a benzaldehyde derivative or a phenyl ketone derivative is a step of obtaining a coupling product of formula (7) or (8) via a reaction step represented by reaction 1-1 or reaction 1-2, and a purification step may be performed after the reaction step.

[0067] (Reaction 1-1) [ka] (Reaction 1-2) [ka]

[0068] The polyoxyethylene derivative used in the coupling reaction is a compound represented by formula (16), in which P 1 The detailed explanation of and w is the same as above, and X 2 is an amino group, a carboxy group, an active ester group, an active carbonate group, a formyl group, a maleimidyl group, a hydroxy group, an alkylene group, an azide group, or E 2 and E 2 is a leaving group, and the leaving group is not particularly limited as long as it is a group having reactivity in a coupling reaction, and examples thereof include a chloro group, a bromo group, an iodo group, a mesylate group, a tosylate group, and a chloromethanesulfonate group, with a mesylate group or a tosylate group being preferred, and a mesylate group being more preferred.

[0069] In formulas (17) and (18), R 6,R 7 , R 8 , R 9 , R 11 , R 13 and R 14 The detailed description of is the same as above, and X 3 isX 2 There is no particular limitation as long as it reacts with the group to form a covalent bond, but specifically, the group may be an amino group, a carboxy group, a thiol group, a hydroxyl group, an alkylene group, an azide group, or an E 2 and E 2 is the same as above, and X 2 and X 3 Preferred combinations are those shown in Table 1.

[0070] [Table 1]

[0071] For example, in the formula (16) of reaction 1-1 and reaction 1-2, X 2 E 2 In this case, a compound represented by formula (16) and a compound represented by formula (17) or formula (18) 3 is a hydroxy group in an aprotic solvent such as toluene, benzene, xylene, acetonitrile, ethyl acetate, diethyl ether, t-butyl methyl ether, tetrahydrofuran, chloroform, dichloromethane, dimethyl sulfoxide, dimethylformamide, or dimethylacetamide, or in the presence of an organic base such as triethylamine, N-methylmorpholine, potassium t-butoxide, or sodium hexamethyldisilazide, or an inorganic base such as potassium carbonate, potassium hydroxide, or sodium hydride. The proportions of the benzaldehyde derivative or phenyl ketone derivative, organic base, and inorganic base used are not particularly limited, but are preferably equimolar or more relative to the chemically reactive functional group of the compound of formula (16). An organic base may also be used as a solvent.

[0072] In the formula (16) of reaction 1-1 and reaction 1-2, X 2 E2 In other cases, X in Table 1 2 and X 3 The reaction conditions for the above combination can be found in many general textbooks, and can be carried out according to, for example, "Greg T. Hermanso, Bioconjugate Techniques, 3rd ed."

[0073] After the reaction steps of Reaction 1-1 and Reaction 1-2, impurities produced as by-products in the reaction, compounds remaining in the reaction without being consumed, and the base catalyst are preferably removed in a purification step. The purification method is not particularly limited, and purification can be performed by extraction, recrystallization, adsorption treatment, reprecipitation, column chromatography, supercritical extraction, or the like.

[0074] Here, between the coupling step and the acetalization step, a dialkylation step may be provided in which the coupling product of formula (7) or (8) is reacted with a monohydric alcohol under acidic conditions to obtain a dialkyl acetal structure represented by formula (9) or (10), and the acetalization step may be carried out using the dialkyl acetal structure represented by formula (9) or (10) obtained in the dialkylation step instead of the coupling product of formula (7) or (8).

[0075] Monohydric alcohols are R 21 -OH, R 22 -OH or R 21 -OH and R 22 Alcohols containing -OH, R 16 and R 17 are each independently a hydrocarbon group having 1 to 10 carbon atoms, preferably a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, and a t-butyl group, more preferably a methyl group, an ethyl group, and a propyl group, and even more preferably a methyl group and an ethyl group.

[0076] The reaction of the coupling product represented by formula (7) or formula (8) with a monohydric alcohol is carried out in an aprotic solvent such as toluene, benzene, xylene, acetonitrile, ethyl acetate, diethyl ether, t-butyl methyl ether, tetrahydrofuran, chloroform, dichloromethane, dimethyl sulfoxide, dimethylformamide, or dimethylacetamide, or in the absence of a solvent, in the presence of an acid catalyst. The acid catalyst may be either an organic acid or an inorganic acid, and is not particularly limited, but specific examples include p-toluenesulfonic acid, pyridinium p-toluenesulfonate, methanesulfonic acid, 10-camphorsulfonic acid, hydrogen chloride, iodine, ammonium chloride, oxalic acid, and boron trifluoride diethyl ether complex.

[0077] Furthermore, a dehydrating agent may be added to the reaction system in order to eliminate water molecules produced in the reaction. The type of dehydrating agent is not particularly limited as long as it does not interfere with the reaction, and examples of the dehydrating agent include orthoesters such as methyl orthoformate, sodium sulfate, magnesium sulfate, alumina, silica gel, and molecular sieves, and preferably orthoesters or molecular sieves.

[0078] After the reaction of the coupling product represented by formula (7) or (8) with the monohydric alcohol, it is preferable to remove impurities produced as by-products in the reaction, compounds remaining in the reaction without being consumed, and the base catalyst in a purification step. The purification method is not particularly limited, and purification can be performed by extraction, recrystallization, adsorption treatment, reprecipitation, column chromatography, supercritical extraction, or the like.

[0079] After the coupling step, the coupling product represented by formula (7) or (8) is subjected to an acidic condition to be reacted with a hydroxymethyl group at the 2-position and a substituent (-CH=CB 1 ) m C(R 1 )(R 2 )-OH(B 1 , m, R 1 , R 2The acetalization step of reacting a phenol having the formula (11) with a phenol having the formula (12) (wherein the formula (11) is as described above) is a step of obtaining an acetal structure represented by formula (20), (21), (23) or (24) through a reaction step represented by any one of reactions 2-1 to 2-4, and a purification step may be performed after the reaction step.

[0080] (Reaction 2-1) [ka] (Reaction 2-2) [ka] (Reaction 2-3) [ka] (Reaction 2-3) [ka]

[0081] In formulas (19) to (24), P 1 , L 1 , B 1 , m, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 and R 14 The details are the same as those in the above description of the amino group-containing acetal-type releasable polyoxyethylene derivative.

[0082] Reactions 2-1 to 2-4 are processes in which the coupling product represented by formula (7) or formula (8) is reacted with the phenol represented by formula (19) or formula (22) in an aprotic solvent such as toluene, benzene, xylene, acetonitrile, ethyl acetate, diethyl ether, t-butyl methyl ether, tetrahydrofuran, chloroform, dichloromethane, dimethyl sulfoxide, dimethylformamide, or dimethylacetamide, or in the absence of a solvent, in the presence of an acid catalyst to obtain an acetal structure of formula (20), (21), (23), or formula (24).

[0083] The acid catalyst may be either an organic acid or an inorganic acid, and is not particularly limited, but specific examples include p-toluenesulfonic acid, pyridinium p-toluenesulfonate, methanesulfonic acid, 10-camphorsulfonic acid, hydrogen chloride, iodine, ammonium chloride, oxalic acid, boron trifluoride diethyl ether complex, etc. Furthermore, a dehydrating agent may be added to the reaction system in order to remove water molecules generated in the reaction, and the type of the dehydrating agent is not particularly limited as long as it does not interfere with the reaction, and examples of the dehydrating agent include sodium sulfate, magnesium sulfate, alumina, silica gel, and molecular sieves, and molecular sieves are preferred.

[0084] After the reaction steps of Reactions 2-1 to 2-4, impurities produced as by-products in the reaction, compounds remaining in the reaction without being consumed, and base catalyst are preferably removed in a purification step. The purification method is not particularly limited, but the product can be purified by extraction, recrystallization, adsorption treatment, reprecipitation, column chromatography, supercritical extraction, or the like.

[0085] In particular, between the acetalization step and the step of introducing a leaving group structure to the hydroxy group, a step of deprotecting the amino group protected by a protecting group in the coupling product of formulas (7) and (8) or the dialkyl acetal structure of formulas (9) and (10) and a step of introducing a functional group capable of reacting with a biofunctional molecule to the deprotected amino group after the deprotection step may be provided. The conditions for the deprotection step of the protecting group and the step of introducing a functional group capable of reacting with a biofunctional molecule to the deprotected amino group can be found in many general textbooks, and the deprotection step can be carried out, for example, according to "Wuts, PGM; Greene, TW Protective Groups in Organic Synthesis, 4th ed.; Wiley-Interscience: New York, 2007", and the step of introducing a functional group capable of reacting with a biofunctional molecule to the deprotected amino group can be carried out, for example, according to "Greg T. Hermanso, Bioconjugate Techniques, 3rd ed."

[0086] The leaving group structure introduction step of introducing a leaving group structure in place of a terminal hydroxy group after the acetalization step or the step of introducing a functional group capable of reacting with a biofunctional molecule into the deprotected amino group is a step of converting a hydroxy group in the polyoxyethylene derivative represented by formula (20), (21), (23), or formula (24) into a succinimidyloxycarbonyloxy group, a phthalimidyloxycarbonyloxy group, a 4-nitrophenoxycarbonyloxy group, a 1-imidazolylcarbonyloxy group, a pentafluorophenoxycarbonyloxy group, a benzotriazol-1-yloxycarbonyloxy group, or a 7-azabenzotriazol-1-yloxycarbonyloxy group, and a purification step may be performed after the reaction step.

[0087] The conversion of the hydroxyl group is performed by condensing the polyoxyethylene derivatives represented by formulas (20), (21), (23) and (24) with the reagents for converting the polyoxyethylene derivatives to the respective leaving groups shown in Table 1 in an aprotic solvent such as toluene, benzene, xylene, acetonitrile, ethyl acetate, diethyl ether, t-butyl methyl ether, tetrahydrofuran, chloroform, dichloromethane, dimethyl sulfoxide, dimethylformamide or dimethylacetamide, or in the presence of an organic base such as triethylamine, N-methylmorpholine, pyridine or 4-dimethylaminopyridine, or an inorganic base such as sodium carbonate, sodium hydrogencarbonate, sodium acetate or potassium carbonate in the absence of a solvent. The proportions of the reagents and base catalysts shown in Table 1 are not particularly limited, but are preferably equimolar or more relative to the hydroxyl group of the polyoxyethylene derivatives of formulas (20), (21), (23) or (24). The reagents shown in Table 2 may be commercially available products or may be produced using known reactions.

[0088] [Table 2]

[0089] After the reaction step in the leaving group structure introduction step, impurities produced as by-products in the reaction, compounds remaining in the reaction without being consumed, and a base catalyst are preferably removed in a purification step. The purification method is not particularly limited, and purification can be performed by extraction, recrystallization, adsorption treatment, reprecipitation, column chromatography, supercritical extraction, or the like.

[0090] The acetal-type releasable polyoxyethylene conjugate of the present invention is an acetal-type releasable polyethylene glycol derivative having -OC(O)E 1 The compound is obtained by reacting a group with an amino group contained in a biofunctional molecule, and is represented by the following formula (9), (10), (11) or (12).

[0091] [ka] [ka] [ka] [ka]

[0092] In formulas (11), (12), (13) and (14), B 2 is a hydrogen atom or -C(R 1 )(R 2 )OC(O)NHD 1 and D 1 is a residue obtained by removing an amino group constituting a carbamate bond from an amino group contained in a biofunctional molecule, P 2 is a polyoxyethylene derivative portion or a conjugate between a polyoxyethylene derivative portion and a biofunctional molecule, w is an integer from 1 to 8; R 1 , R 2 , R 3 , R 4 , R 5 and R 12 is a hydrocarbon group having 1 to 10 carbon atoms or a hydrogen atom, R 6 represents an optionally substituted hydrocarbon group having 1 to 24 carbon atoms or a hydrogen atom, R 7 , R 8 , R 9 , R 10 and R 11 are each independently an electron-withdrawing substituent, an electron-donating substituent, or a hydrogen atom, R 13 represents an optionally substituted hydrocarbon group having 1 to 10 carbon atoms or a hydrogen atom, R 14 represents an optionally substituted hydrocarbon group having 1 to 10 carbon atoms, R13 is R 14 and m is 0 or 1.

[0093] In formulas (11) to (14), R 1 ~R 12 , L 1 The details of and m are the same as those described above. In formulas (11) to (14), B 2 is a hydrogen atom or -C(R 1 )(R 2 )OC(O)NHD 1 and preferably a hydrogen atom.

[0094] In formulas (11) to (14), D 1 is a residue of amino groups contained in a biofunctional molecule excluding the amino groups constituting a carbamate bond, and the carbamate bond may be formed independently from each of the multiple amino groups contained in the biofunctional molecule. For example, -OC(O)E of the acetal-type releasable polyoxyethylene derivative containing the amino group 1 When the group forms a carbamate bond with one of the amino groups contained in the biofunctional molecule, in the acetal-type releasable polyoxyethylene conjugate containing the amino group, P 2 is one, and when it forms a carbamate bond with two, P 2 There are two.

[0095] Biofunctional molecules are not particularly limited, but are substances involved in the diagnosis, cure, mitigation, treatment or prevention of diseases in humans or other animals. Specific examples include proteins, peptides, nucleic acids, cells, viruses, etc., and suitable proteins or peptides include hormones, cytokines, antibodies, aptamers, enzymes, etc.

[0096] More specifically, cytokines include interferon type I, type II, and type III, which regulate immunity, interleukins, tumor necrosis factors, and their receptor antagonists. Growth factors include erythropoietin, which is a hematopoietic factor, and granulocyte colony-stimulating factor (GCSF), which is a stimulating factor. Blood coagulation factors include factor V, factor VII, factor VIII, factor IX, factor X, and factor XII. Hormones include calcitonin, insulin, its analogs, exenatide, GLP-1, somatostatin, and human growth hormone. Antibodies include full-length antibodies, and antibody fragments such as Fab and svFV. Aptamers include DNA aptamers and RNA aptamers. Enzymes include superoxide dismutase and uricase.

[0097] Suitable proteins include interferons, interleukins, erythropoietin, GCSF, Factor VIII, Factor IX, human growth hormone, antibody fragments, and the like, more preferably human growth hormone, interferon, GCSF, erythropoietin, or antibody fragments (particularly Fabs).

[0098] Suitable peptides include insulin, bivalirudin, teriparatide, exenatide, enfuvirtide, degarelix, mifamurtide, nesiritide, goserelin, glatiramer, octreotide, lanreotide, icatibant, zicotinide, pramlintide, romiplostim, calcitonin, oxytocin, leuprorelin, and glucagon, and more preferably, insulin, exenatide, and calcitonin (especially salmon calcitonin).

[0099] In formulas (9) to (12), P 2 is a polyoxyethylene derivative portion, or a conjugate of a polyoxyethylene derivative portion and a biofunctional molecule. The detailed description of the polyoxyethylene derivative part is given below.1 is synonymous with.

[0100] The conjugate of the polyoxyethylene derivative portion and the biofunctional molecule is a conjugate of the acetal-type releasable polyoxyethylene derivative portion. 1 X 1 D 2 D 2 is a group formed by the reaction of a functional group capable of reacting with a biofunctional molecule with the biofunctional molecule. The details of the functional group capable of reacting with a biofunctional molecule are as follows: X of the acetal-type releasable polyoxyethylene derivative 1 The term "biofunctional molecule" is synonymous with the description of the above, and is not particularly limited, but is a substance involved in the diagnosis, cure, mitigation, treatment or prevention of a disease in humans or other animals. Specifically, it includes proteins, peptides, nucleic acids, cells, viruses, etc., and suitable proteins or peptides include hormones, cytokines, antibodies, aptamers, enzymes, etc.

[0101] D 2 Suitable examples of the biofunctional molecules are target-directed biofunctional molecules, such as antibodies and aptamers. Examples of antibodies include antibody fragments such as Fab, Fab', and F(ab'). 2 The aptamer includes a peptide aptamer, an RNA aptamer, and a DNA aptamer.

[0102] The method for producing the acetal-type releasable polyoxyethylene conjugate containing an amino group of the present invention will be described. To produce the polyoxyethylene conjugate of the present invention, a coupling step is carried out in which the acetal-type polyoxyethylene derivative and a biofunctional molecule are reacted in a neutral or basic buffer solution which may contain a water-soluble organic solvent such as acetonitrile, dimethyl sulfoxide, or N,N-dimethylformamide to obtain the polyoxyethylene conjugate, and a purification step is carried out after the coupling step in which the unreacted polyoxyethylene derivative, the biofunctional molecule, or by-products are removed under basic conditions. The term "neutral or basic" refers to a pH of 6.5 to 11.0, preferably a pH of 7.0 to 10.5, more preferably a pH of 7.0 to 10.0, and particularly preferably a pH of 7.0 to 9.0.

[0103] A neutral or basic buffer solution is an aqueous solution that has a buffering effect that mitigates the effects of adding small amounts of acid or base from outside, or changing the concentration by diluting the solution, thereby maintaining the pH (hydrogen ion exponent) at a nearly constant neutral or basic level.

[0104] A water-soluble organic solvent can also be added to the buffer solution. In this case, examples of the water-soluble organic solvent include methanol, ethanol, propanol, isopropanol, tetrahydrofuran, acetone, acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, triethylamine, pyridine, and hexamethylphosphoric acid triamide, preferably methanol, ethanol, propanol, isopropanol, tetrahydrofuran, acetone, acetonitrile, dimethyl sulfoxide, and N,N-dimethylformamide, and more preferably acetonitrile, dimethyl sulfoxide, and N,N-dimethylformamide.

[0105] Specific examples of the purification method carried out after the coupling step include ion exchange chromatography, gel filtration chromatography, hydrophobic interaction chromatography, reverse phase chromatography, and affinity chromatography.

[0106] In the purification step, the unreacted polyoxyethylene derivative, biofunctional molecule or by-product can be removed under neutral or basic conditions under which the acetal structure in the acetal-type releasable polyoxyethylene conjugate is not easily hydrolyzed, and the neutral or basic conditions are pH 6.5 to pH 11.0, preferably pH 7.0 to 10.5, more preferably pH 7.0 to 10.0, and particularly preferably pH 7.0 to 9.0. EXAMPLES

[0107] The present invention will be described in more detail below based on examples, but the present invention is not limited to the following examples.

[0108] The following examples were obtained: 1 H-NMR was obtained from JNM-ECZ400 or JNM-ECA600 manufactured by JEOL Ltd. A φ5 mm tube was used for the measurement, and the deuterated solvent was D 2 O or CDCl containing tetramethylsilane (TMS) as an internal standard 3 , d6-DMSO was used. The molecular weight and terminal functional group purity of the obtained acetal-type releasable polyoxyethylene conjugate were calculated using liquid chromatography (GPC and HPLC). The liquid chromatography systems used were "HLC-8320GPC EcoSEC" manufactured by Tosoh Corporation for GPC and "Nexera" manufactured by Shimadzu Corporation for HPLC.

[0109] (Example 1-1) A three-neck flask was charged with NaH (dispersion in paraffin liquid, 21.8 mg, 0.50 mmol) and THF (2.5 g), and then N-Methyl-N-(2-hydroxyethyl)-4-aminobenzaldehyde (17.9 mg, 0.1 mmol) and NOF Corp. SUNBRIGHT ME-200MS (α-methyl-ω-[(methylsulfonyl)oxy]poly(oxyethylene), 500 mg, 0.025 mmol) were dissolved therein, and the reaction was carried out at 60°C for 6 hours under a nitrogen atmosphere. After quenching by adding 25% aqueous ammonium chloride solution (10 mL), extraction was carried out using dichloromethane (10 mL), and the organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated, dissolved in ethyl acetate, and hexane was added to carry out crystallization. The crystals obtained by filtration were dried under reduced pressure to obtain the compound of formula (25).

[0110] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 3.10(3H,s,-NC H 3 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -OC H 2 C H 2 NCH 3 -), 6.73(2H, d, arom.H), 7.72(2H, d, arom.H), 9.73(1H, s, -CO H ) [ka]

[0111] (Example 1-2) In a three-neck flask, the compound of formula (25) (300 mg, 0.015 mmol), 2,6-bis(hydroxymethyl)-p-cresol (151.4 mg, 0.900 mmol), tetrahydrofuran (1.5 g) and BHT (0.3 mg) were charged and dissolved, and then molecular sieve 5A (300 mg) and pyridinium p-toluenesulfonate (6.8 mg, 0.027 mmol) were added and reacted for 4 hours at 40 ° C under a nitrogen atmosphere. N-methylmorpholine (5.5 mg, 0.054 mmol) was added and stirred for 5 minutes at 25 ° C under a nitrogen atmosphere, and then filtered. The filtrate was diluted with ethyl acetate, and hexane was added to perform crystallization, and the crystals obtained by filtration were dried under reduced pressure to obtain the compound of formula (26).

[0112] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 2.30(3H, s, -C H 3 ), 3.01(3H, s, - NC H 3 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -OC H 2 C H 2 NCH 3 -), 4.66(1H, dd, -OC H 2 OH), 4.76(1H, dd, -OC H 2 OH), 4.93(1H, d, -C H 2 O-), 5.14(1H, d, -C H 2 O-), 5.92(1H, s, -C H<),6.73(2H, d, arom.H), 6.87(1H, s, arom.H), 7.03(1H, s, arom.H), 7.40(2H, d, arom.H) [ka]

[0113] (Examples 1-3) The compound of formula (26) (200 mg, 0.010 mmol), BHT (0.2 mg) and toluene (0.8 g) were charged into a three-neck flask, and after dissolution, triethylamine (15 mg, 0.15 mmol) and p-nitrophenyl chloroformate (20.2 mg, 0.1 mmol) were charged and reacted for 3 hours at 60°C under a nitrogen atmosphere. After the reaction, the mixture was diluted with a mixed solvent of ethyl acetate and acetonitrile to which BHT had been added, and hexane was added to perform crystallization, followed by suction filtration to obtain crystals. The obtained crystals were dried under reduced pressure to obtain the compound of formula (27).

[0114] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 2.30(3H, s, -C H 3 ), 3.01(3H, s, - NC H 3 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -OC H 2 C H 2 NCH 3 -), 4.93(1H, d, -C H 2 O-), 5.14(1H, d, -C H 2O-), 5.34(2H, dd, -OC H 2 OCO-), 5.92(1H, s, -C H <), 6.53(2H, d, arom.H), 6.73(2H, d, arom.H), 6.87(1H, s, arom.H), 7.03(1H, s, arom.H), 7.40(2H, d, arom.H), 7.59(2H, d, arom.H) Number average molecular weight (Mn): 20,532 [ka]

[0115] (Example 2-1) In a three-neck flask, NaH (dispersion in paraffin liquid, 21.8 mg, 0.50 mmol) and THF (2.5 g) were charged, and then 4-(4-hydroxypiperidin-1-ly)benzaldehyde (41.0 mg, 0.2 mmol) and NOF Corp.'s SUNBRIGHT ME-200MS (α-methyl-ω-[(methylsulfonyl)oxy]poly(oxyethylene), 500 mg, 0.025 mmol) were dissolved, and the reaction was carried out at 60°C for 5 hours. After quenching by adding 25% ammonium chloride aqueous solution (10 mL), extraction was carried out using dichloromethane (10 mL), and the organic layer was dried over anhydrous sodium sulfate and filtered. After concentration, the mixture was dissolved in ethyl acetate, and hexane was added to carry out crystallization. The crystals obtained by filtration were dried under reduced pressure to obtain the compound of formula (28).

[0116] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.71(2H, ddt, -C H 2 -), 1.99(2H, ddt, -C H 2 -), 3.21(2H,ddd,-NC H 2-), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -C H <, -NC H 2 -), 6.91(2H, d, arom.H), 7.73(2H, d, arom.H), 9.76(1H, s, -CO H ) [ka]

[0117] (Example 2-2) In a three-neck flask, the compound of formula (28) (300 mg, 0.015 mmol), 2,6-bis(hydroxymethyl)-p-cresol (151.4 mg, 0.900 mmol), tetrahydrofuran (1.5 g) and BHT (0.3 mg) were charged and dissolved, and then molecular sieve 5A (300 mg) and pyridinium p-toluenesulfonate (56.5 mg, 0.225 mmol) were added and reacted for 4 hours at 40 ° C under a nitrogen atmosphere. N-methylmorpholine (45.5 mg, 0.45 mmol) was added and stirred for 5 minutes at 25 ° C under a nitrogen atmosphere, and then filtered. The filtrate was diluted with ethyl acetate, and hexane was added to perform crystallization, and the crystals obtained by filtration were dried under reduced pressure to obtain the compound of formula (29).

[0118] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.71 (2H, ddt, -C H 2 -), 1.99(2H, ddt, -C H 2 -), 2.30(3H, s, -C H3 ), 2.99(2H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -C H <, -NC H 2 -), 4.66(1H, dd, -OC H 2 OH), 4.76(1H, dd, -OC H 2 OH), 4.93(1H, d, -C H 2 O-), 5,14(1H, d, -C H 2 O-), 5,93(1H, s, -C H <), 6.76(1H, s, arom.H), 6.95(2H, d, arom.H), 7.02(1H, s, arom.H), 7.43(2H, d, arom.H) [ka]

[0119] (Example 2-3) The compound of formula (29) (200 mg, 0.010 mmol), BHT (0.2 mg) and toluene (0.8 g) were charged into a three-neck flask, dissolved, triethylamine (15 mg, 0.15 mmol) and p-nitrophenyl chloroformate (20.2 mg, 0.1 mmol) were charged, and the reaction was carried out for 3 hours at 60°C under a nitrogen atmosphere. After the reaction, the mixture was diluted with a mixed solvent of ethyl acetate and acetonitrile to which BHT had been added, and hexane was added to perform crystallization, followed by suction filtration to obtain crystals. The obtained crystals were dried under reduced pressure to obtain the compound of formula (30).

[0120] 1 H-NMR (CDCl 3 , internal standard TMS); δ (ppm): 1.71 (2H, ddt, -C H 2 -), 1.99 (2H, ddt, -C H 2 -), 2.30 (3H, s, -C H 3 ), 3.21 (2H, ddd, -NC H 2 -), 3.38 (3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52 - 4.23 (m, -(OC H 2 C H 2 ) n -,-C H <,-NC H 2 -), 4.93 (1H, d, -C H 2 O-), 5,14 (1H, d, -C H 2 O-), 5.34 (2H, dd, -OC H 2 OCO-), 5,93 (1H, s, -C H <), 6.53 (2H, d, arom.H), 6.76 (1H, s, arom.H), 6.95 (2H, d, arom.H), 7.02 (1H, s, arom.H), 7.43 (2H, d, arom.H), 7.59 (2H, d, arom.H) Number average molecular weight (Mn): 20,934

Chem.

[0121] (Example 3 - 1) In a three-neck flask, NaH (dispersion in paraffin liquid, 21.8 mg, 0.50 mmol) and THF (2.5 g) were charged, and then 4-[4-(hydroxymethyl)piperidin-1-ly]benzaldehyde (43.8 mg, 0.2 mmol) and NOF Corp. SUNBRIGHT ME-200MS (α-methyl-ω-[(methylsulfonyl)oxy]poly(oxyethylene), 500 mg, 0.025 mmol) were dissolved, and the reaction was carried out at 60°C for 5 hours. After quenching by adding 25% ammonium chloride aqueous solution (10 mL), extraction was carried out using dichloromethane (10 mL), and the organic layer was dried over anhydrous sodium sulfate and filtered. After concentration, the mixture was dissolved in ethyl acetate, crystallized by adding hexane, filtered, and dried under reduced pressure to obtain the compound of formula (31).

[0122] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.33(2H, ddt, -C H 2 -), 1.87(3H, m, -C H 2 -, -C H <), 2.92(2H, ddd, -NC H 2 -), 3.35(2H, ddd,-NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -OC H 2 CH<), 6.90(2H, d, arom.H), 7.73(2H, d, arom.H), 9.75(1H, s, -CO H ) [ka]

[0123] (Example 3-2) In a three-neck flask, the compound of formula (31) (300 mg, 0.015 mmol), 2,6-bis(hydroxymethyl)-p-cresol (151.4 mg, 0.900 mmol), tetrahydrofuran (1.5 g) and BHT (0.3 mg) were charged and dissolved, and then molecular sieve 5A (300 mg) and pyridinium p-toluenesulfonate (56.5 mg, 0.225 mmol) were added and reacted for 4 hours at 40 ° C under a nitrogen atmosphere. N-methylmorpholine (45.5 mg, 0.45 mmol) was added and stirred for 5 minutes at 25 ° C under a nitrogen atmosphere, and then filtered. The filtrate was diluted with ethyl acetate, and hexane was added to perform crystallization, and the crystals obtained by filtration were dried under reduced pressure to obtain the compound of formula (32).

[0124] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.33(2H, ddt, -C H 2 -), 1.87(3H, m, -C H 2 -, -C H <), 2.30(3H, s, -C H 3 ), 2.76(2H, ddd, -NC H 2 -), 3.35(2H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -OC H 2 CH<), 4.67(2H, dd, -OC H2 OH), 4.93(1H, d, -C H 2 O-), 5.14(1H, d, -C H 2 O-), 5,93(1H, s, -C H <), 6.76(1H, s, arom.H), 6.95(2H, d, arom.H), 7.02(1H, s, arom.H), 7.43(2H, d, arom.H) [ka]

[0125] (Example 3-3) The compound of formula (32) (200 mg, 0.010 mmol), BHT (0.2 mg) and toluene (0.8 g) were charged into a three-neck flask, dissolved, triethylamine (15 mg, 0.15 mmol) and p-nitrophenyl chloroformate (20.2 mg, 0.1 mmol) were charged, and the reaction was carried out for 3 hours at 60°C under a nitrogen atmosphere. After the reaction, the mixture was diluted with a mixed solvent of ethyl acetate and acetonitrile to which BHT had been added, and hexane was added to perform crystallization, followed by suction filtration to obtain crystals. The obtained crystals were dried under reduced pressure to obtain the compound of formula (33).

[0126] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.33(2H, ddt, -C H 2 -), 1.87(3H, m, -C H 2 -, -C H <), 2.30(3H, s, -C H 3 ), 2.76(2H, ddd, -NC H 2 -), 3.35(2H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -OC H 2 CH<), 4.93(1H, d, -C H 2 O-), 5,14(1H, d, -C H 2 O-), 5.34(2H, dd, -OC H 2 OCO-), 5.93(1H, s, -C H <), 6.53(2H, d, arom.H), 6.76(1H, s, arom.H), 6.95(2H, d, arom.H), 7.02(1H, s, arom.H), 7.43(2H, d, arom.H),7.59(2H, d, arom.H) Number average molecular weight (Mn): 20,121

change

[0127] (Example 4-1) In a three-neck flask, 4-(piperazin-1-yl)benzaldehyde (38.1 mg, 0.2 mmol) synthesized with reference to patent document CN105777632A, SUNBRIGHT ME-200HS (500 mg, 0.025 mmol) manufactured by NOF Corp., and chloroform (5.0 g) were charged and dissolved, and then triethylamine (2.5 mg, 0.025 mmol) was charged and reacted for 2 hours at 40°C under a nitrogen atmosphere. After the reaction, the mixture was washed with acid water using a 5% NaCl / 6N HCl aqueous solution, and then washed with alkali water using a 5% NaCl aqueous solution (pH 12), and the organic layer was dried over anhydrous sodium sulfate and filtered. After concentration, the mixture was dissolved in ethyl acetate, and hexane was added to perform crystallization, followed by suction filtration to obtain crystals. The obtained crystals were dried under reduced pressure to obtain a compound of formula (34).

[0128] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, tt, -C H 2 -), 1.63(2H, tt, -C H 2 -), 1.69(2H, dd, -C H 2 -), 2.38(2H, t, -C H 2 CO-), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 6.92(2H, d, arom.H), 7.78(2H, d, arom.H), 9.81(1H, s, -CO H ) [ka]

[0129] (Example 4-2) In a three-neck flask, the compound of formula (34) (300 mg, 0.015 mmol), 2,6-bis(hydroxymethyl)-p-cresol (151.4 mg, 0.900 mmol), tetrahydrofuran (1.5 g), and BHT (0.3 mg) were charged and dissolved, and then molecular sieve 5A (300 mg) and pyridinium p-toluenesulfonate (56.5 mg, 0.225 mmol) were added, and the reaction was carried out at 40 ° C. for 4 hours under a nitrogen atmosphere. N-methylmorpholine (45.5 mg, 0.45 mmol) was added, and the mixture was stirred at 25 ° C. for 5 minutes under a nitrogen atmosphere, and then filtered. The filtrate was diluted with ethyl acetate, and hexane was added to perform crystallization, and the crystals obtained by filtration were dried under reduced pressure to obtain the compound of formula (35).

[0130] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H,tt,-C H 2 -), 1.63(2H,tt,-C H 2 -), 1.69(2H,dd,-C H 2 -), 2.30(3H, s, -C H 3 ), 2.38(2H,t, -C H 2 CO-), 3.20(2H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.67(2H, dd, -OC H 2OH), 4.93(1H, d, -C H 2 O-), 5,14(1H, d, -C H 2 O-), 5.95(1H, s, -C H <), 6.77(1H, s, arom.H), 6.96(2H, d, arom.H), 7.03(1H, s, arom.H), 7.48(2H, d, arom.H) [ka]

[0131] (Example 4-3) The compound of formula (35) (200 mg, 0.010 mmol), BHT (0.2 mg) and toluene (0.8 g) were charged into a three-neck flask, dissolved, triethylamine (15 mg, 0.15 mmol) and p-nitrophenyl chloroformate (20.2 mg, 0.1 mmol) were charged, and the reaction was carried out for 3 hours at 60°C under a nitrogen atmosphere. After the reaction, the mixture was diluted with a mixed solvent of ethyl acetate and acetonitrile to which BHT had been added, and hexane was added to perform crystallization, followed by suction filtration to obtain crystals. The obtained crystals were dried under reduced pressure to obtain the compound of formula (36).

[0132] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, tt, -C H 2 -), 1.63(2H, tt, -C H 2 -), 1.69(2H, dd, -C H 2 -), 2.38(2H, t, -C H 2 CO-), 2.20(2H, ddd, -NC H 2 -), 2.30(3H, s, -C H 3 ), 3.38(3H, s, -(OCH2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.93(1H, d, -C H 2 O-), 5.14(1H, d, -C H 2 O-), 5.34(2H, dd, -OC H 2 OCO-), 5.95(1H, s, -C H <), 6.77(1H, s, arom.H), 6.53(2H, d, arom.H), 6.96(2H, d, arom.H), 7.03(1H, s, arom.H), 7.48(2H, d, arom.H), 7.59(2H, d, arom.H) Number average molecular weight (Mn): 20,634

change

[0133] (Example 5-1) In a three-neck flask, 3-Fluoro-4-piperazin-1-yl-benzaldehyde (41.7 mg, 0.2 mmol) synthesized with reference to patent document CN105777632A, SUNBRIGHT ME-200HS (500 mg, 0.025 mmol) manufactured by NOF Corp., and chloroform (5.0 g) were charged and dissolved, and then triethylamine (2.5 mg, 0.025 mmol) was charged and reacted for 2 hours at 40°C under a nitrogen atmosphere. After the reaction, the mixture was washed with acid water using a 5% NaCl / 6N HCl aqueous solution, and then washed with alkali water using a 5% NaCl aqueous solution (pH 12), and the organic layer was dried over anhydrous sodium sulfate and filtered. After concentration, the mixture was dissolved in ethyl acetate, and hexane was added to perform crystallization, followed by suction filtration to obtain crystals. The obtained crystals were dried under reduced pressure to obtain a compound of formula (37).

[0134] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -C H 2 -), 2.38(2H, t, -COC H 2 -), 3.24(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 6.99(1H, dd, arom.H), 7.54-7.61(2H, m, arom.H), 9.85(1H, s, -CO H ) [ka]

[0135] (Example 5-2) In a three-neck flask, the compound of formula (37) (300 mg, 0.015 mmol), 2,6-bis(hydroxymethyl)-p-cresol (151.4 mg, 0.900 mmol), tetrahydrofuran (1.5 g) and BHT (0.3 mg) were charged and dissolved, and then molecular sieve 5A (300 mg) and pyridinium p-toluenesulfonate (13.6 mg, 0.054 mmol) were added, and the reaction was carried out for 4 hours at 40 ° C. under a nitrogen atmosphere. N-methylmorpholine (45.5 mg, 0.45 mmol) was added, and the mixture was stirred for 5 minutes at 25 ° C. under a nitrogen atmosphere, and then filtered. The filtrate was diluted with ethyl acetate (90 g), and hexane (60 g) was added to perform crystallization, followed by filtration and drying under reduced pressure to obtain the compound of formula (38).

[0136] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H,dd, -C H 2 -), 1.63-1.68(4H, m, -C H 2 -), 2.30(3H, s, -C H 3 ), 2.38(2H, t, -COC H 2 -), 3.08(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.67(2H, dd, -OC H 2 OH), 4.93(1H, d, -CH 2 O-), 5,14(1H, d, -C H 2 O-), 5.94(1H, s, -C H <), 6.77(1H, s, arom.H), 6.97(1H, dd, arom.H), 7.06(1H, s, arom.H), 7.53-7.60(2H, m, arom.H) [ka]

[0137] (Example 5-3) The compound of formula (38) (200 mg, 0.010 mmol), BHT (0.2 mg) and toluene (0.8 g) were charged into a three-neck flask, dissolved, triethylamine (15 mg, 0.15 mmol) and p-nitrophenyl chloroformate (20.2 mg, 0.1 mmol) were charged, and the reaction was carried out for 3 hours at 60°C under a nitrogen atmosphere. After the reaction, the mixture was diluted with a mixed solvent of ethyl acetate and acetonitrile to which BHT had been added, and hexane was added to perform crystallization, followed by suction filtration to obtain crystals. The obtained crystals were dried under reduced pressure to obtain the compound of formula (39).

[0138] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -C H 2 -), 2.30(3H, s, -C H 3 ), 2.38(2H, t, -COC H 2 -), 3.08(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.93(1H, d, -C H 2 O-), 5,14(1H, d, -C H 2 O-), 5.34(2H, dd, -OC H 2 OCO-), 5.94(1H, s, -C H <), 6.77(1H, s, arom.H), 6.99(1H, d, arom.H), 6.53(2H, d, arom.H), 7.03(1H, s, arom.H), 7.53-7.60(4H, m, arom.H) Number average molecular weight (Mn): 21,243 [ka]

[0139] (Example 6-1) In a 50 ml three-neck flask equipped with a thermometer, a nitrogen inlet tube and a stirrer, 2-(Piperazin-1-yl)benzaldehyde (38.1 mg, 0.2 mmol) synthesized with reference to patent document CN105777632A, SUNBRIGHT ME-200HS (500 mg, 0.025 mmol) manufactured by NOF Corp., and chloroform (5.0 g) were charged and dissolved, and then triethylamine (2.5 mg, 0.025 mmol) was charged and reacted for 2 hours at 40°C under a nitrogen atmosphere. After the reaction, the mixture was washed with acid water using a 5% NaCl / 6N HCl aqueous solution, and then washed with alkali water using a 5% NaCl aqueous solution (pH 12), after which the organic layer was dried over anhydrous sodium sulfate and filtered. After concentration, the mixture was dissolved in ethyl acetate, and hexane was added to perform crystallization, followed by suction filtration to obtain crystals. The obtained crystals were dried under reduced pressure to obtain a compound of formula (40).

[0140] 1 H-NMR (CDCl 3 , internal standard TMS); δ (ppm): 1.42 (2H, tt, -C H 2 -), 1.63 - 1.68 (4H, m, -C H 2 -), 2.38 (2H, t, -C H 2 CO-), 3.07 (4H, ddd, -NC H 2 -), 3.38 (3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52 - 4.23 (m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 7.15 (1H, d, arom.H), 7.18 (1H, dd, arom.H), 7.55 (1H, dd, arom.H), 7.83 (1H, d, arom.H), 10.35 (1H, s, -CO H )

Chem.

[0141] (Example 6 - 2) In a three-neck flask, the compound of formula (40) (300 mg, 0.015 mmol), 2,6-bis(hydroxymethyl)-p-cresol (151.4 mg, 0.900 mmol), tetrahydrofuran (1.5 g) and BHT (0.3 mg) were charged and dissolved, and then molecular sieve 5A (300 mg) and pyridinium p-toluenesulfonate (56.5 mg, 0.225 mmol) were added and reacted at 40 ° C for 4 hours under a nitrogen atmosphere. N-methylmorpholine (45.5 mg, 0.45 mmol) was added and stirred at 25 ° C for 5 minutes under a nitrogen atmosphere, and then filtered. The filtrate was diluted with ethyl acetate, crystallized by adding hexane, filtered, and dried under reduced pressure to obtain the compound of formula (41).

[0142] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, tt, -C H 2 -), 1.63-1.68(4H, m, -C H 2 -), 2.30(3H, s, -C H 3 ), 2.38(2H, t, -C H 2 CO-), 2.93(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.63(2H, dd, -OC H 2 OH), 4.97(1H, d, -C H 2 O-), 5,17(1H, d, -C H 2O-), 6.44(1H, s, -C H <), 6.81(1H, s, arom.H), 7.05(1H, s, arom.H), 7.16(1H, d, arom.H), 7.25(1H, dd, arom.H), 7.42(1H, dd, arom.H), 7.76(1H, d, arom.H) [ka]

[0143] (Example 6-3) The compound of formula (41) (200 mg, 0.010 mmol), BHT (0.2 mg) and toluene (0.8 g) were charged into a three-neck flask, dissolved, triethylamine (15 mg, 0.15 mmol) and p-nitrophenyl chloroformate (20.2 mg, 0.1 mmol) were charged, and the reaction was carried out for 3 hours at 60°C under a nitrogen atmosphere. After the reaction, the mixture was diluted with a mixed solvent of ethyl acetate and acetonitrile to which BHT had been added, and hexane was added to perform crystallization, followed by suction filtration to obtain crystals. The obtained crystals were dried under reduced pressure to obtain the compound of formula (42).

[0144] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, tt, -C H 2 -), 1.63-1.68(4H, m, -C H 2 -), 2.30(3H, s, -C H 3 ), 2.38(2H, t, -C H 2 CO-), 2.93(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H2 C H 2 ) n -, -NC H 2 -), 4.97(1H, d, -C H 2 O-), 5,17(1H, d, -C H 2 O-), 5.34(2H, dd, -OC H 2 OCO-), 6.44(1H, s, -C H <), 6.53(2H, d, arom.H), 6.81(1H, s, arom.H), 7.05(1H, s, arom.H), 7.16(1H, d, arom.H), 7.25(1H, dd, arom.H), 7.42(1H, dd, arom.H), 7.59(2H, d, arom.H), 7.76(1H, d, arom.H) Number average molecular weight (Mn): 20,983 [ka]

[0145] (Comparative Example 1-1) Three flasks were charged with 3-hydroxybenzaldehyde (2.00g, 16.4mmol), trimethyl orthoformate (3.48g, 32.8mmol), and methanol (17g), and p-toluenesulfonic acid monohydrate (0.312g, 1.64mmol) was added and reacted at 25°C for 2 hours. Sodium hydroxide was added and stirred for a while, and then the solvent was distilled off under reduced pressure. The residue was dissolved in dichloromethane and washed with 5wt% aqueous sodium bicarbonate solution and 25wt% saline solution in that order, and the organic layer was dried over anhydrous sodium sulfate. After filtration, the solvent was distilled off under reduced pressure to obtain the compound of formula (43).

[0146] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 3.33(6H, s, -OCH3), 5.35(1H, s, -CH<), 6.81(1H, d, arom.H ), 6.95(1H, d, arom.H), 7.23-7.26(1H, m, arom.H) [ka]

[0147] (Comparative Example 1-2) In a three-neck flask, 2,4-di(hydroxymethyl)phenol (50.0 mg, 0.324 mmol) synthesized according to the literature (Freeman, JH; JAm. Chem. Soc. 1952, 74, 6257-6260), the compound of formula (43) (217 mg, 1.29 mmol), 2,6-di-tert-butyl-p-cresol (7.14 mg, 0.0324 mmol), anhydrous sodium sulfate (1 g) and cyclopentyl methyl ether (10 g) were charged, and p-toluenesulfonic acid monohydrate (4.10 mg, 0.0212 mmol) was added and reacted at 40 ° C for 2 hours. N-methylmorpholine was added and stirred for a while, and then filtration was performed. After washing with 10 wt% saline, the organic layer was dried over anhydrous sodium sulfate. After filtration, the solvent was distilled off under reduced pressure to obtain the compound of formula (44).

[0148] 1 H-NMR (d6-DMSO, internal standard TMS); δ(ppm): 4.42(2H, d, -C H 2 OH), 4.93(1H, d, -C H 2 O-), 5.10(1H, t, -CH 2 O H ), 5.15(1H, d, -C H 2 O-), 6.01(1H, s, -C H <), 6.80-7.21(7H, m, arom.H ), 9.53(1H, bs, >CO H ) [ka]

[0149] (Comparative Example 1-3) In a three-neck flask, the compound of formula (44) (37.0 mg, 0.141 mmol), ME-200MS (α-methyl-ω-[(methylsulfonyl)oxy]poly(oxyethylene), 705 mg, 0.0353 mmol) manufactured by NOF Corp., potassium carbonate (97.0 mg, 0.705 mmol) and acetonitrile (3.5 g) were charged and reacted at 80°C for 4 hours. After filtration, the solvent was distilled off under reduced pressure and the residue was dissolved in dichloromethane. After washing with 10 wt% saline, the organic layer was dried over anhydrous sodium sulfate. After filtration, the solvent was distilled off under reduced pressure and the residue was dissolved in toluene (50 g). Hexane (50 g) was added to perform crystallization, and the mixture was filtered and dried under reduced pressure to obtain the compound of formula (45).

[0150] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 3.38(3H, s, -OC H 3 ), 3.52-4.18(m, -(OC H 2 C H 2 ) n -), 4.62(2H, s, -C H 2 OH), 4.98(1H, d, -C H 2 O-), 5.18(1H, d, -C H 2 O-), 5.95(1H, s, -C H <), 6.87-7.34(7H, m, arom.H ) [ka]

[0151] (Comparative Examples 1-4) In a 50mL three-neck flask equipped with a thermometer, a nitrogen inlet tube, a stirrer and a cooling tube, the compound of formula (45) (300mg, 0.0150mmol), di(N-succinimidyl)carbonate (46.0mg, 0.180mmol), triethylamine (21.0mg, 0.208mmol) and dichloromethane (5g) were charged and reacted at 25℃ for 12 hours. After filtration, the mixture was washed with 5wt% saline, and the organic layer solvent was distilled off under reduced pressure. The residue was dissolved in ethyl acetate (6g), dried over anhydrous sodium sulfate and then filtered. After adding ethyl acetate (44g), hexane (50g) was added to cause crystallization, filtered and dried under reduced pressure to obtain the compound of formula (46).

[0152] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 2.85(4H, s, -COC H 2 C H 2 CO-), 3.38(3H, s, -OC H 3 ), 3.52-4.18(m, -(OC H 2 C H 2 ) n -), 5.00(1H, d, -C H 2 OCH<), 5.18(1H, d, -C H 2 O-), 5.25(2H, s, -CH 2 OCO-), 5.97(1H, s, -C H <), 6.96-7.35(7H, m, arom.H ) [ka]

[0153] Example 7 β-alanine was dissolved in 0.1M sodium phosphate buffer solution (pH 8.5) to prepare a 20mg / mL β-alanine solution. The compound of formula (36) (50mg, 0.0025mmol) was dissolved in 20mg / mL β-alanine solution (1.5mL) and reacted for 6 hours at 25°C under a nitrogen atmosphere. After the reaction, the mixture was diluted with 0.1M sodium phosphate buffer solution (pH 8.5) containing 20% ​​by weight of sodium chloride, and extracted with chloroform (3g). The organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was diluted with ethyl acetate, and hexane was added to perform crystallization. The crystals obtained by filtration were dried under reduced pressure to obtain the compound of formula (47).

[0154] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, tt, -C H 2 -), 1.63(2H, tt, -C H 2 -), 1.69(2H, dd, -C H 2 -), 2.30(3H, s, -C H 3 ), 2.38(2H, t, -C H 2 CO-), 2.53(2H, t, -C H 2 COOH), 3.20(2H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.37-3.43(2H, m, -C H 2 NH-), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.93(1H, d, -C H 2O-), 5.08(1H, dd, -C H 2 OCONH-), 5.14(1H, d, -C H 2 O-), 5.38(1H, dd, -C H 2 OCONH-), 5.95(1H, s, -C H <), 6.77(1H, s, arom.H), 6.96(2H, d, arom.H), 7.03(1H, s, arom.H), 7.48(2H, d, arom.H) [ka]

[0155] Example 8 β-alanine was dissolved in 0.1M sodium phosphate buffer solution (pH 8.5) to prepare a 20mg / mL β-alanine solution. The compound of formula (40) (50mg, 0.0025mmol) was dissolved in 20mg / mL β-alanine solution (1.5mL) and reacted for 6 hours at 25°C under a nitrogen atmosphere. After the reaction, the mixture was diluted with 0.1M sodium phosphate buffer solution (pH 8.5) containing 20% ​​by weight of sodium chloride, and extracted with chloroform (3g). The organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was diluted with ethyl acetate, and hexane was added to perform crystallization. The crystals obtained by filtration were dried under reduced pressure to obtain the compound of formula (48).

[0156] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -C H 2 -), 2.30(3H, s, -C H 3 ), 2.38(2H, t, -COC H 2 -), 2.53(2H, t, -C H2 COOH), 3.08(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.93(1H, d, -C H 2 O-), 5.08(1H, dd, -C H 2 OCONH-), 5,14(1H, d, -C H 2 O-), 5.38(1H, dd, -C H 2 OCONH-), 5.94(1H, s, -C H <), 6.77(1H, s, arom.H), 6.99(1H, d, arom.H), 7.03(1H, s, arom.H), 7.53-7.60(2H, m, arom.H) [ka]

[0157] Example 9 β-alanine was dissolved in 0.1M sodium phosphate buffer solution (pH 8.5) to prepare a 20mg / mL β-alanine solution. The compound of formula (42) (50mg, 0.0025mmol) was dissolved in 20mg / mL β-alanine solution (1.5mL) and reacted for 6 hours at 25°C under a nitrogen atmosphere. After the reaction, the mixture was diluted with 0.1M sodium phosphate buffer solution (pH 8.5) containing 20% ​​by weight of sodium chloride and extracted with chloroform (3g). The organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was diluted with ethyl acetate, and hexane was added to perform crystallization. The crystals obtained by filtration were dried under reduced pressure to obtain the compound of formula (49).

[0158] 1 H-NMR(CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, tt, -C H 2 -), 1.63 - 1.68(4H, m, -C H 2 -), 2.30(3H, s, -C H 3 ), 2.38(2H, t, -C H 2 CO-), 2.53(2H, t, -C H 2 COOH), 2.93(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52 - 4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.97(1H, d, -C H 2 O-), 5.08(1H, dd, -C H 2 OCONH-), 5,17(1H, d, -C H 2 O-), 5.38(1H, dd, -OC H 2 OCONH-), 6.44(1H, s, -C H <), 6.81(1H, s, arom.H), 7.05(1H, s, arom.H), 7.16(1H, d, arom.H), 7.25(1H, dd, arom.H), 7.42(1H, dd, arom.H), 7.76(1H, d, arom.H)

Chem.

[0159] Comparative Example 2 A 20 mg / mL buffer solution of β-alanine was prepared using 0.1 M sodium phosphate buffer solution (pH 8.5). The compound of formula (46) (143 mg, 0.0072 mmol) and a 20 mg / mL buffer solution of β-alanine (3.0 g) were charged into a 50 mL three-neck flask equipped with a thermometer, a nitrogen inlet tube, and a stirrer, and after dissolution, the reaction was carried out at 25°C for 6 hours. After dissolving salt (750 mg), extraction was carried out using chloroform (4.5 g), and the organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was diluted with ethyl acetate (45 g), and hexane (33 g) was added to perform crystallization, filtered, and dried under reduced pressure to obtain the compound of formula (50).

[0160] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 2.47(2H,t,-C H 2 COOH), 3.38(3H, s, -(OCH 2 CH 2 ) n O.C. H 3 ), 3.37-3.43(2H, m, -C H 2 NH-), 3.52-4.21(m, -(OC H 2 C H 2 ) n -), 4.89(1H, d, -C H 2 O-), 5.05(1H, d, -C H 2 O-), 5.14(2H, d d, -C H 2 OCONH-), 6.22(1H, s, -C H <), 6.52-7.57(7H, arom.H ) [ka]

[0161] (Degradability test 1) The compounds (2 mg) of formula (47), formula (48), formula (49), and formula (50) were each dissolved in phosphate buffered saline (2 mL) at pH 7.4, and then allowed to stand in a thermostatic bath at 37°C, and HPLC measurement was performed at any timing. Figure 1 shows the amount of β-alanine adduct at any timing when the amount of β-alanine adduct at 0 hours of standing is taken as 100%. The approximation equation of the exponential approximation curve obtained from the graph, with the y-intercept set at 100, is y=100e -ax (-a is the power exponent.)

number

[0162] (Example 10-1) The compound of formula (51) was obtained in the same manner as in Example 6-1, except that 2-Bromo-4-(Piperazin-1-yl)benzaldehyde (53.8 mg, 0.2 mmol) was used instead of 2-(Piperazin-1-yl)benzaldehyde.

[0163] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2-), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 3.24(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 6.68(1H, dd, arom.H), 7.04(1H, d, arom.H),7.85(1H, m, arom.H), 10.15(1H, s, -CO H )

[0164] [ka]

[0165] (Example 10-2) The compound of formula (52) was obtained in the same manner as in Example 6-2, except that the compound of formula (51) (300 mg, 0.015 mmol) was used instead of the compound of formula (40).

[0166] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COCH 2 -), 3.24(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.67(2H, dd, -C H 2 OH), 5.25(2H,dd,-CH-OC H 2 -CH<), 6.25(1H,s,-C H <), 6.8-7.7 (5H, aromatic H)

[0167] [ka]

[0168] (Example 10-3) The compound of formula (53) was obtained in the same manner as in Example 6-3, except that the compound of formula (52) (200 mg, 0.010 mmol) was used instead of the compound of formula (41).

[0169] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 3.24(4H, ddd, -NC H 2-), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.67(2H, dd, -C H 2 OH), 5.25(2H,dd,-CH-OC H 2 -CH<), 6.25(1H,s,-C H <),6.8-7.7(5H,arom.H),8.29(2H,d,arom.H) Number average molecular weight (Mn): 21,392

[0170] [ka]

[0171] (Example 11-1) The compound of formula (54) was obtained in the same manner as in Example 6-1, except that 2-chloro-4-(piperazin-1-yl)benzaldehyde (44.9 mg, 0.2 mmol) was used instead of 2-(piperazin-1-yl)benzaldehyde.

[0172] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 3.24(4H, ddd, -NCH 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 6.79(2H, m, arom.H),7.85(1H, m, arom.H), 10.24(1H, s, -CO H )

[0173] [ka]

[0174] (Example 11-2) The compound of formula (55) was obtained in the same manner as in Example 6-2, except that the compound of formula (54) (300 mg, 0.015 mmol) was used instead of the compound of formula (40).

[0175] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 3.24(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H2 C H 2 ) n -, -NC H 2 -), 4.67(2H, dd, -C H 2 OH), 5.25(2H,dd,-CH-OC H 2 -CH<), 6.25(1H,s,-C H <), 6.8-7.7 (5H, aromatic H)

[0176] [ka]

[0177] (Example 11-3) The compound of formula (56) was obtained in the same manner as in Example 6-3, except that the compound of formula (55) (200 mg, 0.010 mmol) was used instead of the compound of formula (41).

[0178] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 3.24(4H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H2 -), 4.67(2H, dd, -C H 2 OH), 5.25(2H,dd,-CH-OC H 2 -CH<), 6.25(1H,s,-C H <),6.7-7.8(5H,arom.H),8.29(2H,d,arom.H) Number average molecular weight (Mn): 20,174

[0179] [ka]

[0180] (Example 12-1) The compound of formula (57) was obtained in the same manner as in Example 6-1, except that 2-Chloro-4-(methyl(2-(methylamino)ethyl)amino)benzaldehyde (45.3 mg, 0.2 mmol) was used instead of 2-(Piperazin-1-yl)benzaldehyde.

[0181] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 2.75(3H, s, >NC H 3 ), 2.94(3H, s, >NC H 3 ), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OCH 2 C H 2 ) n -, -N(CH 3 ) C H 2 -C H 2 -N(CH 3 )-), 6.79(2H, m, arom.H),7.85(1H, m, arom.H), 10.33(1H, s, -CO H )

[0182] [ka]

[0183] (Example 12-2) The compound of formula (58) was obtained in the same manner as in Example 6-2, except that the compound of formula (57) (300 mg, 0.015 mmol) was used instead of the compound of formula (40).

[0184] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 2.75(3H, s, >NC H 3 ), 2.94(3H, s, >NC H 3 ), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 CH 2 ) n -, -N(CH 3 ) C H 2 -C H 2 -N(CH 3 )-),4.67(2H, dd, -C H 2 OH), 5.25(2H,dd,-CH-OC H 2 -CH<), 6.25(1H,s,-C H <), 6.8-7.5(5H,arom.H)

[0185] [ka]

[0186] (Example 12-3) The compound of formula (59) was obtained in the same manner as in Example 6-3, except that the compound of formula (58) (200 mg, 0.010 mmol) was used instead of the compound of formula (41).

[0187] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 2.75(3H, s, >NC H 3 ), 2.94(3H, s, >NC H 3 ), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -N(CH 3 ) C H 2 -C H 2 -N(CH 3 )-),4.67(2H, dd, -C H 2 OH), 5.25(2H,dd,-CH-OC H 2 -CH<), 6.25(1H,s,-C H <),6.8-7.5(5H,arom.H),8.3(2H,d,arom.H) Number average molecular weight (Mn): 20,590

[0188] [ka]

[0189] (Example 13-1) The compound of formula (60) was obtained in the same manner as in Example 6-1, except that 3-Fluoro-4-(methyl(2-(methylamino)ethyl)amino)benzaldehyde (42.1 mg, 0.2 mmol) was used instead of 2-(Piperazin-1-yl)benzaldehyde.

[0190] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2-), 2.75(3H, s, >NC H 3 ), 2.94(3H, s, >NC H 3 ), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -N(CH 3 ) C H 2 -C H 2 -N(CH 3 )-), 6.79(2H, m, arom.H),7.85(1H, m, arom.H), 10.21(1H, s, -CO H )

[0191] [ka]

[0192] (Example 13-2) The compound of formula (61) was obtained in the same manner as in Example 6-2, except that the compound of formula (60) (300 mg, 0.015 mmol) was used instead of the compound of formula (40).

[0193] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 2.75(3H, s, >NC H3 ), 2.94(3H, s, >NC H 3 ), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -N(CH 3 ) C H 2 -C H 2 -N(CH 3 )-),4.67(2H, dd, -C H 2 OH), 5.25(2H,dd,-CH-OC H 2 -CH<), 6.25(1H,s,-C H <), 6.5-7.7 (5H, aromatic H)

[0194] [ka]

[0195] (Example 13-3) The compound of formula (62) was obtained in the same manner as in Example 6-3, except that the compound of formula (61) (200 mg, 0.010 mmol) was used instead of the compound of formula (41).

[0196] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2-), 2.75(3H, s, >NC H 3 ), 2.94(3H, s, >NC H 3 ), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -N(CH 3 ) C H 2 -C H 2 -N(CH 3 )-),4.67(2H, dd, -C H 2 OH), 5.25(2H,dd,-CH-OC H 2 -CH<), 6.25(1H,s,-C H <),6.5-7.7(5H,arom.H),8.29(2H,d,arom.H) Number average molecular weight (Mn): 21,029

[0197] [ka]

[0198] Example 14 The compound of formula (63) was obtained in the same manner as in Example 9, except that the compound of formula (53) (50 mg, 0.0025 mmol) was used instead of the compound of formula (42).

[0199] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, tt, -C H 2 -), 1.63(2H, tt, -C H 2-), 1.69(2H, dd, -C H 2 -), 2.30(3H, s, -C H 3 ), 2.38(2H, t, -C H 2 CO-), 2.53(2H, t, -C H 2 COOH), 3.20(2H, ddd, -NC H 2 -), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.37-3.43(2H, m, -C H 2 NH-), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.93(1H, d, -C H 2 O-), 5.08(1H, dd, -C H 2 OCONH-), 5.14(1H, d, -C H 2 O-), 5.38(1H, dd, -C H 2 OCONH-), 5.95(1H, s, -C H <), 6.6-7.5(5H, arom.H)

[0200] [ka]

[0201] Example 15 The compound of formula (64) was obtained in the same manner as in Example 9, except that the compound of formula (56) (50 mg, 0.0025 mmol) was used instead of the compound of formula (42).

[0202] 1 H-NMR (CDCl 3 , internal standard TMS); δ (ppm): 1.42 (2H, tt, -C H 2 -), 1.63 (2H, tt, -C H 2 -), 1.69 (2H, dd, -C H 2 -), 2.30 (3H, s, -C H 3 ), 2.38 (2H, t, -C H 2 CO-), 2.53 (2H, t, -C H 2 COOH), 3.20 (2H, ddd, -NC H 2 -), 3.38 (3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.37 - 3.43 (2H, m, -C H 2 NH-), 3.52 - 4.23 (m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.93 (1H, d, -C H 2 O-), 5.08 (1H, dd, -C H 2 OCONH-), 5.14 (1H, d, -C H 2 O-), 5.38 (1H, dd, -C H 2 OCONH-), 5.95 (1H, s, -C H <), 6.6 - 7.5 (5H, arom.H)

[0203]

Chemical formula

[0204] Example 16 Compound (65) was obtained in the same manner as in Example 9, except that compound (59) (50 mg, 0.0025 mmol) was used instead of compound (42).

[0205] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 2.75(3H, s, >NC H 3 ), 2.94(3H, s, >NC H 3 ), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -N(CH 3 ) C H 2 -C H 2 -N(CH 3 )-),4.67(2H, dd, -C H 2 OH), 5.25(2H,dd,-CH-OC H 2 -CH<), 6.25(1H,s,-C H <), 6.6-7.8 (5H, aromatic H)

[0206] [ka]

[0207] (Example 17) Compound (66) was obtained in the same manner as in Example 9, except that compound (62) (50 mg, 0.0025 mmol) was used instead of compound (42).

[0208] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, dd, -C H 2 -), 1.63-1.68(4H, m, -CH 2 C H 2 C H 2 -), 2.38(2H, t, -COC H 2 -), 2.75(3H, s, >NC H 3 ), 2.94(3H, s, >NC H 3 ), 3.38(3H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -N(CH 3 ) C H 2 -C H 2 -N(CH 3 )-),4.67(2H, dd, -C H 2 OH), 5.25(2H,dd,-CH-OC H 2 -CH<), 6.25(1H,s,-C H <), 6.6-7.5(5H,arom.H)

[0209] [ka]

[0210] (Example 18-1) The compound of formula (67) was obtained in the same manner as in Example 4-1, except that SUNBRIGHT GL2-400GC2 (500 mg, 0.0125 mmol) manufactured by NOF Corporation was used instead of SUNBRIGHT ME-200HS manufactured by NOF Corporation.

[0211] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, tt, -C H 2 -), 1.63(2H, tt, -C H 2 -), 1.69(2H, dd, -C H 2 -), 2.38(2H, t, -C H 2 CO-), 3.38(6H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 6.92(2H, d, arom.H), 7.78(2H, d, arom.H), 9.85(1H, s, -CO H )

[0212] [ka]

[0213] (Example 18-2) The compound of formula (68) was obtained in the same manner as in Example 4-2, except that the compound of formula (67) (300 mg, 0.0075 mmol) was used instead of the compound of formula (34).

[0214] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H,tt,-C H 2 -), 1.63(2H,tt,-C H 2 -), 1.69(2H,dd,-C H 2 -), 2.30(3H, s, -C H 3 ), 2.38(2H,t, -C H 2 CO-), 3.20(2H, ddd, -NC H 2 -), 3.38(6H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.67(2H, dd, -OC H 2 OH), 4.93(1H, d, -C H 2 O-), 5,14(1H, d, -C H 2 O-), 5.95(1H, s, -C H <), 6.3-7.5(6H, arom.H)

[0215] [ka]

[0216] (Example 18-3) The compound of formula (69) was obtained in the same manner as in Example 4-3, except that the compound of formula (68) (200 mg, 0.005 mmol) was used instead of the compound of formula (35).

[0217] 1 H-NMR (CDCl 3 , internal standard TMS); δ(ppm): 1.42(2H, tt, -C H 2 -), 1.63(2H, tt, -C H 2 -), 1.69(2H, dd, -C H 2 -), 2.38(2H, t, -C H 2 CO-), 2.20(2H, ddd, -NC H 2 -), 2.30(3H, s, -C H 3 ), 3.38(6H, s, -(OCH 2 CH 2 )nOC H 3 ), 3.52-4.23(m, -(OC H 2 C H 2 ) n -, -NC H 2 -), 4.93(1H, d, -C H 2 O-), 5.14(1H, d, -C H 2 O-), 5.34(2H, dd, -OC H 2 OCO-), 5.95(1H, s, -C H <), 6.3-7.9(10H, arom.H) Number average molecular weight (Mn): 39,217

[0218] [ka]

[0219] (Degradability test 2) The compounds (2 mg) of formula (63), formula (64), formula (65), and formula (66) were each dissolved in phosphate buffered saline (2 mL) of pH 7.4, and then allowed to stand in a thermostatic bath at 37°C, and HPLC measurement was performed at any timing. Figure 2 shows the amount of β-alanine adduct at any timing when the amount of β-alanine adduct at 0 hours of standing is taken as 100%. The approximation equation of the exponential approximation curve obtained from the graph, with the y-intercept set to 100, is y=100e -ax The half-lives (t1 / 2) calculated from the formula (Math 1) based on (-a is a power exponent) were 19.9 hours for the compound of formula (63), 19.6 hours for the compound of formula (64), 2.7 hours for the compound of formula (65), and 21.6 hours for the compound of formula (66). That is, the compounds of the present invention represented by formulas (63), (64), (65), and (66) were shown to have the intended half-life under physiological conditions, compared with the half-life of 231 days (as shown in Degradation Test 1) of the compound exemplified in Patent Document 3 represented by formula (50), which is a prior invention.

[0220] (Example 19) IL2 (NKMAX) represented by SEQ ID NO:1 was dissolved in 20 mM bicarbonate buffer (pH 9.0) to a final concentration of 0.43 mg / mL, and compound (36) was dissolved in a final concentration of 8.5 mg / mL, and the reaction was carried out at 4° C. for 3 hours. Then, a 20 mg / mL glycine solution was added and stirred at 25° C. for 1 hour. After stirring, the solution was exchanged with 1 mM Tris-HCl (pH 9.0) by ultrafiltration, and anion exchange chromatography was carried out under the following conditions. The target fraction was collected and concentrated to obtain (RLS)PEG(20k)-IL2.

[0221] HPLC equipment: Nexera (Shimadzu Corporation) Column: Asahipack ES-502N 7C (7.5x100mm, 9um, Showa Denko) Flow rate: 1mL / min Column temperature: 15℃ Injection volume: 50μL Detector: PDA (measurement wavelength: 280nm) Mobile phase A: 1mM Tris-HCl buffer (pH9.0) Mobile phase B: 1mM Tris-HCl buffer (pH9.0) containing 0.1M NaCl Gradient program (A / B): 100 / 0 (0min) → 100 / 0 (20min) → 75 / 25 (25min) → 0 / 100 (30min) → 0 / 100 (35min)

[0222] Comparative Example 3 IL2 (manufactured by NKMAX) represented by SEQ ID NO:1 was dissolved in 20 mM bicarbonate buffer (pH 9.0) to a final concentration of 0.43 mg / mL, and SUNBRIGHT MENP-20T (manufactured by NOF Corp.) was dissolved in 20 mM bicarbonate buffer (pH 9.0) to a final concentration of 6.5 mg / mL, and the mixture was reacted at 25° C. for 3 hours. Then, a 20 mg / mL glycine solution was added and stirred at 25° C. for 1 hour. After stirring, the solution was exchanged with 1 mM Tris-HCl (pH 9.0) by ultrafiltration, and anion exchange chromatography was performed in the same manner as in Example 18, and the target fraction was collected and concentrated to obtain (NRL)PEG(20k)-IL2.

[0223] (Cell proliferation test) Viable cell concentration was 1.89 x 10 5 90μL of CTLL-2 (RCB0637, Riken BRC) cells / mL was seeded into each well of a 96-well plate (Corning) and cultured for 4 hours. Then, 10μL of 0-20ng / mL (RLS)PEG(20k)-IL2 in BSA-containing PBS solution or 0-20ng / mL (NRL)PEG(20k)-IL2 in BSA-containing PBS solution was added to each well. After addition, the plate was cultured for 45 hours. Then, 100μL of Cell Titor Glo reagent (Promega) was added to each well, and the amount of luminescence by luciferase was measured.

[0224] The maximum luminescence obtained with (RLS)PEG(20k)-IL2 or (NRL)PEG(20k)-IL2 was taken as 100% cell proliferation rate, and the cell proliferation rate at each concentration was calculated for (RLS)PEG(20k)-IL2 and (NRL)PEG(20k)-IL2. Figure 4 shows the proliferation rate at each concentration of (RLS)PEG(20k)-IL2 and (NRL)PEG(20k)-IL2. The regression curve equation obtained from the graph is

number

Claims

1. A compound represented by the following formula (1), formula (2), formula (3) or formula (4): Acetal-type releasable polyoxyethylene derivative containing a .alpha. group. 【Chemistry 1】 【Chemistry 2】 【Chemistry 3】 【Chemistry 4】 (In formulas (1), (2), (3) and (4), B １ is a hydrogen atom or -C(R １ ) (R ２ ) O.C.(O)E １ and E １ is a leaving group, P １ is a polyoxyethylene derivative moiety, w is an integer from 1 to 8; R １ , R ２ , R ３ , R ４ , R ５ and R １２ Each independently represents a carbon atom having 1 to 10 carbon atoms. is a hydrogen group or a hydrogen atom, R ６ represents an optionally substituted hydrocarbon group having 1 to 24 carbon atoms or a hydrogen atom, R ７ , R ８ , R ９ , R １０ and R １１ are each independently an electron-withdrawing substituent, is a donor substituent or a hydrogen atom, R １３ represents an optionally substituted hydrocarbon group having 1 to 10 carbon atoms or a hydrogen atom, R １４ represents an optionally substituted hydrocarbon group having 1 to 10 carbon atoms, R １３ is R １４ and m is 0 or 1.

2. m is 0, R １ and R ２ is a hydrogen atom, R ３ , R ４ , R ５ and R １２ Gaso 2. The amino acid of claim 1, wherein each of the amino groups is independently a hydrogen atom or a methyl group. Acetal-type releasable polyoxyethylene derivatives containing a group.

3. m is 0, R １ and R ２ is a hydrogen atom, R ３ , R ４ , R ５ and R １２ Gaso each independently represents a hydrogen atom or a methyl group; L １ R in １３ and R １４ and are combined the law of nature, R １３ is a hydrocarbon group represented by the following formula (5), R １４ is expressed by the following formula (6): The amino group-containing acetal according to claim 1 or 2, characterized in that the hydrocarbon group is A type releasable polyoxyethylene derivative. 【Chemistry 5】 (In formula (5), R １５ and R １６ each independently represents a carbon atom having 1 to 4 carbon atoms which may be substituted is a hydrogen group or a hydrogen atom, q1 is an integer from 1 to 4, (*1) represents the point of attachment to N, (*2) is R １４ represents the point of attachment to 【Chemistry 6】 (In formula (6), R １７ and R １８ each independently represents a carbon atom having 1 to 4 carbon atoms which may be substituted is a hydrogen group or a hydrogen atom, R １９ is CR ２０ or a nitrogen atom, R ２０ represents an optionally substituted hydrocarbon group having 1 to 4 carbon atoms or a hydrogen atom, q2 is an integer from 0 to 3, Z １ is R １９ Carbon or nitrogen atom of P １ A single bond or divalent spacer connecting - and (*3) represents the point of attachment to N, (*4) is R １３ represents the point of attachment to (*5) is P １ represents the point of attachment to

4. m is 0, R １ and R ２ is a hydrogen atom, R ３ , R ４ , R ５ and R １２ are each independently a hydrogen atom or a methyl group, R １３ and R １４ is bonded, R １５ and R １６ is a hydrogen atom, q1 is 2, R １７ and R １８ is a hydrogen atom, R １９ is a methine group or a nitrogen atom, The acetal-type alkylene compound containing an amino group according to claim 3, characterized in that q2 is 2. -subable polyoxyethylene derivatives.

5. The acetal type releasable polyoxyethylene having an amino group according to claim 1, 2, 3 or 4. A method for producing a diethylene derivative, comprising the steps of: A polyoxyethylene derivative and a benzaldehyde derivative or a phenyl ketone derivative By coupling, a coupling represented by the following formula (7) or the following formula (8) is obtained. a coupling step to obtain the coupling product; After the coupling step, a compound represented by the following formula (7) or (8) is reacted under acidic conditions. The coupling product is then treated with a hydroxymethyl group at the 2-position and a hydroxymethyl group at the 4-position or 6-position. Substituent (-CH=CB １ ) C(R １ ) (R ２ )-OH(B １ , m, R １ , R ２ As mentioned above, The acetal structure is obtained by reacting the phenol having the aryl group with the phenol having the aryl group. a polymerization step; and After the acetalization step, a leaving group structure (—OC) is added to the terminal of the substituent at the 4-position or 6-position. (O) E １ ) (E １ is a leaving group) The present invention relates to an acetal type releasable polyoxyethylene having an amino group. Method for producing phenyl derivatives. 【Chemistry 7】 【Chemistry 8】 (In formula (7) and formula (8), P １ , w, R ６ , R ７ , R ８ , R ９ , R １０ , R １１ , R １３ and R １４ is as mentioned above.)

6. Between the coupling step and the acetalization step, a compound represented by the formula (7) is added under acidic conditions. ) or reacting the coupling product of formula (8) with a monohydric alcohol. The dialkyl acetal structure represented by the following formula (9) or (10) is obtained by The method of claim 5, further comprising a dialkylation step for obtaining an acetone having an amino group. A method for producing a tar-type releasable polyoxyethylene derivative. 【Chemistry 9】 【Chemistry 10】 (In formulas (9) and (10), R ２１ and R ２２ are each independently a hydrocarbon group having 1 to 10 carbon atoms, P １ , w, R ６ , R ７ , R ８ , R ９ , R １０ , R １１ , R １３ and R １４ As mentioned above It is as follows.)

7. Between the acetalization step and the leaving group structure introduction step, a step of deprotecting the amino group protected by a protecting group in After the deprotection step, a functional group capable of reacting with a biofunctional molecule is introduced into the deprotected amino group. The method for producing the acetal-type arylamine compound according to claim 5, further comprising the step of: A method for producing a soluble polyoxyethylene derivative.

8. Between the acetalization step and the leaving group structure introduction step, a step of deprotecting the amino group protected by a protecting group in the aryl structure; and After the deprotection step, a functional group capable of reacting with a biofunctional molecule is introduced into the deprotected amino group. The method for producing the acetal-type arylamine compound according to claim 6, further comprising the step of: A method for producing a soluble polyoxyethylene derivative.

9. Represented by the following formula (11), formula (12), formula (13) or formula (14), under physiological conditions The acetal type releasable polyoxyethylene having an amino group is cleaved by Len combination. 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】 (In formulas (11), (12), (13) and (14), B ２ is a hydrogen atom or -C(R １ ) (R ２ )OC(O)NHD １ and D １ The amino group in the biofunctional molecule is converted into an amino group that constitutes a carbamate bond. is a residue excluding the group, P ２ The polyoxyethylene derivative portion or the polyoxyethylene derivative portion and a biological function It is a combination of sex molecules, w is an integer from 1 to 8; R １ , R ２ , R ３ , R ４ , R ５ and R １２ are each independently carbon atoms having 1 to 10 is a hydrogen atom or a hydride group, R ６ represents an optionally substituted hydrocarbon group having 1 to 24 carbon atoms or a hydrogen atom, R ７ , R ８ , R ９ , R １０ and R １１ are each independently an electron-withdrawing substituent, is a child-donating substituent or a hydrogen atom, R １３ represents an optionally substituted hydrocarbon group having 1 to 10 carbon atoms or a hydrogen atom, R １４ represents an optionally substituted hydrocarbon group having 1 to 10 carbon atoms, R １３ is R １４ and m is 0 or 1.

10. The acetal-type releasable polyoxyethylene conjugate containing an amino group according to claim 9. A method for producing the acetal-type releasable polyoxyethylene ether of claim 1 or 2, The ethylene derivative and the biofunctional molecule are mixed in a neutral or A coupling step in which the reaction is carried out in a basic buffer solution, and a neutral buffer solution after the coupling step or a purification step under basic conditions. Method for producing releasable polyoxyethylene conjugates.

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