Crosslinked alginate structure

Abstract

By carrying out a crosslinking reaction using alginic acid derivatives represented by the following formula (I) and the following formula (II), new crosslinked alginic acid, crosslinked alginic acid structures, etc. can be provided. Thus, new crosslinked alginic acid, crosslinked alginic acid structures, etc. can be provided.

Description

Technical Field

[0001] The present invention relates to crosslinked alginic acid, alginic acid derivatives for forming the crosslinked alginic acid, and the like. Background Art

[0002] Alginic acid is a bioabsorbable polysaccharide extracted from brown algae such as Lessonia, Macrocystis, Laminaria, Ascophyllum, Darbilla, Ecklonia cava, Arame, and Kombu, and is a linear polymer formed by polymerization of two types of uronic acids, D-mannuronic acid (M) and L-glucuronic acid (G). More specifically, it is a block copolymer obtained by arbitrarily bonding a homopolymer component of D-mannuronic acid (MM component), a homopolymer component of L-glucuronic acid (GG component), and a component (M / G component) obtained by randomly arranging D-mannuronic acid and L-glucuronic acid. Alginic acid is used in a wide range of fields such as food, medicine, cosmetics, fibers, and papermaking.

[0003] Alginic acid alkali metal salts, which are monovalent salts of alginic acid (e.g., sodium alginate, etc.), are water-soluble, but alginic acid alkaline earth metal salts, which are divalent salts (e.g., calcium alginate, etc.), gelate (insolubilize) by metal ion crosslinking. That is, by adding an aqueous solution containing a divalent metal ion (e.g., calcium ion, barium ion, etc.) to an aqueous solution of alginic acid, an alginic acid gel (ion-crosslinked alginic acid) having a three-dimensional network structure formed by ionic crosslinking can be obtained. Attempts have been made to modify or shape alginic acid alkali metal salts suitable for various uses using this property (Patent Documents 1 to 3). On the other hand, it is known that the aforementioned alginic acid gel, for example, by the presence of a chelating agent such as ethylenediaminetetraacetic acid (EDTA), captures divalent metal ions forming ionic crosslinks in the gel, and thus the gel structure is easily destroyed and restored to alginic acid.

[0004] Polysaccharide derivatives having a maleimide group and / or a thiol group as reactive groups for forming chemical crosslinks are known (Patent Documents 4 to 10, Non-Patent Documents 1 to 2).

[0005] Prior Art Documents

[0006] Patent Documents

[0007] [Patent Document 1] Japanese Patent Application Laid-Open No. 2010-209130

[0008] [Patent Document 2] Japanese Patent Application Laid-Open No. 2007-99902

[0009] [Patent Document 3] International Publication No. 2004 / 099259

[0010] [Patent Document 4] Japanese Patent Application Laid-Open No. 2003-516519

[0011] [Patent Document 5] Japanese Patent Application Laid-Open No. 2015-502957

[0012] [Patent Document 6] FR2967678

[0013] [Patent Document 7] International Publication No. 2014 / 058359 [Patent Document 8] Japanese Patent Application Laid-Open No. 2010-512433 [Patent Document 9] International Publication No. 2018 / 151186 [Patent Document 10] International Publication No. 2019 / 189330

[0014] Non-Patent Document

[0015] [Non-Patent Document 1] ACS Nano (2012), 6(6), 4796-4805.

[0016] [Non-Patent Document 2] Biomacromolecules (2019), 20(6), 2350-2359. Summary of the Invention

[0017] Problems to be Solved by the Invention

[0018] Under the above circumstances, there is a need for crosslinked alginic acid, an alginic acid derivative for forming the crosslinked alginic acid, a crosslinked alginic acid structure, and methods for producing them.

[0019] Means for Solving the Problems

[0020] The present inventors repeatedly conducted in-depth studies to solve the above problems, and as a result, found that by using an alginic acid derivative into which a specified crosslinking group has been introduced (the alginic acid derivatives represented by the following formulas (I) and (II)), crosslinked alginic acid and a crosslinked alginic acid structure (beads / pigmented beads) are formed, and as a result, the beads have high stability and a gel having substance permeability, etc., thus completing the present invention. That is, the present invention is as follows.

[0021] Here, there are provided an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II) shown in the following modes, crosslinked alginic acid obtained by performing a Michael addition reaction using them, and a crosslinked alginic acid structure obtained by performing a Michael addition reaction on a gel obtained by dropping the aforementioned alginic acid derivative into a solution containing a divalent metal ion, and methods for producing the aforementioned alginic acid derivative, crosslinked alginic acid, and crosslinked alginic acid structure.

[0022] That is, the exemplified modes are as described in the following [1] to

[27] .

[0023] [1] Crosslinked alginic acid, which is obtained by carrying out a crosslinking reaction using an alginic acid derivative represented by the following formula (I) and an alginic acid derivative represented by the following formula (II),

[0024] [Chemical formula 1]

[0025]

[0026] [In formula (I), -L 1 -, -NHCO-, and (ALG) are the same as defined in the following first mode],

[0027] [Chemical formula 2]

[0028]

[0029] [In formula (II), -L 2 -, -NHCO-, and (ALG) are the same as defined in the following first mode].

[0030] [2] An alginic acid derivative represented by the following formula (I):

[0031] [Chemical formula 3]

[0032]

[0033] [In formula (I), -L 1 -, -NHCO-, and (ALG) are the same as defined in the following first or second mode].

[0034] [3] The alginic acid derivative represented by formula (I) as described in the foregoing [1] or [2], wherein the introduction rate of the reactive group represented by the following formula (BR-1):

[0035] [Chemical formula 4]

[0036]

[0037] [In formula (BR-1), -L 1 - is the same as defined in the following third mode] is 0.5% to 30% or 1% to 30%.

[0038] [4] The alginic acid derivative represented by formula (I) as described in the foregoing [1] or [2], wherein the weight-average molecular weight of the alginic acid derivative measured by gel filtration chromatography is 100,000 Da to 3,000,000 Da.

[0039] [5] An alginic acid derivative represented by the following formula (II-P):

[0040] [Chemical formula 5]

[0041]

[0042] [In formula (II-P), P 1 -, -L 2 -, -NHCO-, and (ALG) are the same as defined in the following Mode 5].

[0043] [6-1] The alginic acid derivative represented by formula (II) described in the above [1], wherein the following formula (BR-2):

[0044] [Chemical Formula 6]

[0045]

[0046] [In formula (BR-2), -L 2 - is the same as defined in the following Mode 6-1], and the introduction rate of the reactive group is 1.0% to 30%.

[0047] [6-2] The alginic acid derivative represented by formula (II-P) described in the above [5], wherein the following formula (BR-2-P):

[0048] [Chemical Formula 7]

[0049]

[0050] [In formula (BR-2-P), P 1 -, -L 2 - is the same as defined in the following Mode 6-2], and the introduction rate of the reactive group is 1.0% to 30%.

[0051] [7-1] The alginic acid derivative represented by formula (II) described in the above [1], wherein the weight-average molecular weight of the alginic acid derivative measured by gel filtration chromatography is 100,000 Da to 3,000,000 Da.

[0052] [7-2] The alginic acid derivative represented by formula (II-P) described in the above [5], wherein the weight-average molecular weight of the alginic acid derivative measured by gel filtration chromatography is 100,000 Da to 3,000,000 Da.

[0053] [8] The crosslinked alginic acid described in the above [1], wherein the chemical crosslinking has the structure of the following formula (LK-1):

[0054] [Chemical Formula 8]

[0055]

[0056] [In formula (LK-1), the -CONH- and -NHCO- at both ends, -L 1 -, and -L 2 - are the same as defined in the following Mode 8].

[0057] [8-1] Crosslinked alginic acid represented by the following formula (CAL-1):

[0058] [Chemical formula 9]

[0059]

[0060] [In formula (CAL-1), (ALG), -CONH-, -NHCO-, -L 1 -, and -L 2 - have the same definitions as those in [1] above.]

[0061] [8-2] Crosslinked alginic acid represented by the following formula (CAL-1) obtained by the crosslinking reaction described in [1] above:

[0062] [Chemical formula 10]

[0063]

[0064] [In formula (CAL-1), (ALG), -CONH-, -NHCO-, -L 1 -, and -L 2 - have the same definitions as those in [1] above.]

[0065] [9] A method for preparing the crosslinked alginic acid described in [1] above, which comprises adding a solution of an alginic acid derivative represented by formula (I) to a solution of an alginic acid derivative represented by formula (II) to carry out a crosslinking reaction.

[0066]

[10] A method for preparing the crosslinked alginic acid described in [1] above, which comprises adding a solution of an alginic acid derivative represented by formula (II) to a solution of an alginic acid derivative represented by formula (I) to carry out a crosslinking reaction.

[0067]

[11] A method for preparing the crosslinked alginic acid described in [1] above, wherein the chemical crosslinking formed by a Michael addition reaction using an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II) has the structure of the following formula (LK-1):

[0068] [Chemical formula 11]

[0069]

[0070] [In formula (LK-1), the -CONH- and -NHCO- at both ends, -L 1 -, and -L 2 - have the same definitions as those in Mode 11 below.]

[0071]

[12] A crosslinked alginic acid structure, which is obtained by subjecting a gel obtained by dropping a solution of an alginic acid derivative represented by formula (I) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of an alginic acid derivative represented by formula (II). As the crosslinking, it includes ionic crosslinking partially formed by the divalent metal ion and chemical crosslinking formed by a Michael addition reaction.

[0072]

[13] A crosslinked alginic acid structure, which is obtained by subjecting a gel obtained by dropping a solution of an alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of an alginic acid derivative represented by formula (I). As the crosslinking, it includes ionic crosslinking partially formed by the divalent metal ion and chemical crosslinking formed by a Michael addition reaction.

[0073]

[14] A crosslinked alginic acid structure, which is obtained by dropping a solution of a composition containing an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion. As the crosslinking, it includes ionic crosslinking partially formed by the divalent metal ion and chemical crosslinking formed by a Michael addition reaction.

[0074]

[15] The crosslinked alginic acid structure according to any one of the above

[12] to

[14] , wherein the chemical crosslinking formed by the Michael addition reaction using the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) has the structure of the following formula (LK-1):

[0075] [Chemical formula 12]

[0076]

[0077] [In formula (LK-1), the -CONH- and -NHCO- at both ends, -L 1 -, and -L 2 - have the same definitions as those in the 15th aspect described below].

[0078]

[16] The crosslinked alginic acid structure according to any one of the above

[12] to

[15] , which is a fibrous structure, fiber, bead, gel or nearly spherical gel.

[0079]

[17] A medical material, which contains the crosslinked alginic acid structure according to any one of the above

[12] to

[16] .

[0080]

[18] The medical material according to the above

[17] , which is a fibrous structure, fiber, bead, gel or nearly spherical gel.

[0081] 〔19〕A composition comprising an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II).

[0082] 〔19-1〕A composition comprising an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II-P).

[0083] 〔19-2〕A composition comprising an alginic acid derivative selected from an alginic acid derivative represented by formula (I), an alginic acid derivative represented by formula (II), or an alginic acid derivative represented by formula (II-P).

[0084] 〔20〕A method for preparing a crosslinked alginic acid structure, comprising dropping a solution of a composition containing an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion.

[0085] 〔21〕A method for preparing a crosslinked alginic acid structure, comprising: subjecting a gel obtained by dropping a solution of an alginic acid derivative represented by formula (I) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of an alginic acid derivative represented by formula (II), thereby obtaining a crosslinked alginic acid structure containing an ionic crosslink partially formed by a divalent metal ion and a chemical crosslink formed by a Michael addition reaction as crosslinks.

[0086] 〔22〕A method for preparing a crosslinked alginic acid structure, comprising: subjecting a gel obtained by dropping a solution of an alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of an alginic acid derivative represented by formula (I), thereby obtaining a crosslinked alginic acid structure containing an ionic crosslink partially formed by a divalent metal ion and a chemical crosslink formed by a Michael addition reaction as crosslinks.

[0087] 〔23〕The method for preparing a crosslinked alginic acid structure according to any one of the foregoing 〔20〕 to 〔22〕, wherein the chemical crosslink formed by a Michael addition reaction using the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) has the structure of the following formula (LK-1):

[0088] [Chemical formula 13]

[0089]

[0090] [In formula (LK-1), the -CONH- and -NHCO- at both ends, -L 1 -, and -L 2 - are the same as those defined in the following Mode 23].

[0091]

[24] A crosslinked alginic acid structure, which is obtained by subjecting an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II) to ionic crosslinking using a divalent metal ion and chemical crosslinking using a Michael addition reaction, and which has the property of retaining a content.

[0092]

[25] The crosslinked alginic acid according to the above [1] or [8], or the crosslinked alginic acid structure according to any one of the above

[12] to

[16] and

[24] , which has biocompatibility.

[0093] [26a] The alginic acid derivative represented by formula (I) according to the above [2], and the alginic acid derivative represented by formula (II-P) according to the above [5], which have biocompatibility.

[0094]

[27] An amino compound represented by the following formula (AM-2) or a pharmaceutically acceptable salt thereof:

[0095] [Chemical formula 14]

[0096]

[0097] [In formula (AM-2), P 1 , -L 2 - is as defined in the following Mode 27].

[0098] Advantages of the Invention

[0099] The present invention provides a crosslinked alginic acid having a chemical crosslink formed, a crosslinked alginic acid structure, an alginic acid derivative that can be used in the formation of the crosslinked alginic acid and the crosslinked alginic acid structure, and an intermediate (amino compound) for preparing the derivative, etc.

[0100] Preferably, the crosslinked alginic acid and the alginic acid derivative as a raw material are expected to be safe for living organisms.

[0101] In addition, the crosslinking reaction of the alginic acid derivative of the present invention is completed under mild conditions by a Michael addition reaction, and thus it can be used safely and easily.

[0102] The crosslinked alginic acid in some modes is chemically crosslinked by a Michael addition reaction. The crosslinked alginic acid of the present invention can be used in combination with chemical crosslinking and further other crosslinking methods, such as crosslinking using a divalent metal ion using calcium ion, and adjusting the reaction conditions, whereby its stability is improved compared with non-crosslinked alginic acid (for example, monovalent sodium salt of alginic acid) or non-chemically crosslinked alginic acid (for example, crosslinked alginic acid obtained by crosslinking with calcium ion).

[0103] In addition, in some modes, the gel physical properties of the crosslinked product can be adjusted, and the substance permeability can also be adjusted.

[0104] The present invention has at least one or more of these effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0105] Figure 1 A diagram showing the evaluation of the gel stability of the crosslinked alginic acid structure.

[0106] Figure 2 A diagram showing the evaluation of the gel stability of the crosslinked alginic acid structure under EDTA.

[0107] Figure 3 A diagram showing the evaluation of the gel stability of the crosslinked alginic acid structure.

[0108] Figure 4 A diagram showing the evaluation of the gel stability of the crosslinked alginic acid structure under EDTA.

[0109] Figure 5 A diagram showing the evaluation of the gel transmittance of the crosslinked alginic acid structure.

[0110] Figure 6 A diagram showing the evaluation of the gel transmittance of the crosslinked alginic acid structure.

[0111] Figure 7 A diagram showing the evaluation of the biocompatibility of the gel of the crosslinked alginic acid derivative.

[0112] Figure 8 A diagram showing the evaluation of the gel stability of the crosslinked alginic acid structure.

[0113] Figure 9 A diagram showing the evaluation of the gel stability of the crosslinked alginic acid structure under EDTA.

[0114] Figure 10 A diagram showing the evaluation of the gel transmittance of the crosslinked alginic acid structure.

[0115] Figure 11 A diagram showing the evaluation of the biocompatibility of the gel of the crosslinked alginic acid derivative. DETAILED DESCRIPTION OF THE INVENTION

[0116] [Specific embodiments]

[0117] More specifically, the following embodiments [1] to

[24] may be included.

[0118] [1] The first embodiment is as follows. Crosslinked alginic acid is obtained by carrying out a crosslinking reaction using the alginic acid derivative represented by the following formula (I) and the alginic acid derivative represented by the following formula (II).

[0119] [The alginic acid derivative represented by formula (I)] ​​​​​​​​​​​

[0120] An alginic acid derivative represented by the following formula (I):

[0121] [Chemical formula 15]

[0122]

[0123] (In formula (I), (ALG) represents alginic acid; -NHCO- represents an amide bond obtained from any carboxyl group of alginic acid; -L 1 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0124] [Chemical formula 16]

[0125]

[0126] (The hydrogen atom of the methylene group (-CH 2 -) in formulas (L1-1) to (L1-4) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (in the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, R a and R b are each independently a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6substituted by a plurality of groups (e.g., 1 to 10 or 1 to 5) in the alkyl group;

[0127] When two hydrogen atoms of the same methylene group (-CH 2 -) in the formulas (L1-1) to (L1-4) are substituted with C 1～6 alkyl groups, these alkyl groups may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0128] The -NH- group in the formulas (L1-3) and (L1-4) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0129] n is an integer from 1 to 18;

[0130] m is an integer from 1 to 9;

[0131] j is an integer from 0 to 9)).

[0132] [Alginic acid derivative represented by formula (II)]

[0133] An alginic acid derivative represented by the following formula (II):

[0134] [Chemical formula 17]

[0135]

[0136] (In formula (II), (ALG) represents alginic acid; -NHCO- represents an amide bond obtained through an arbitrary carboxyl group of alginic acid; -L 2 - is a linking group selected from the following partial structural formulas [excluding the outside of the dotted lines at both ends]:

[0137] [Chemical formula 18]

[0138]

[0139] (The hydrogen atom of the methylene group (-CH 2 -) in the formulas (L2-1) to (L2-6) may be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (Ra R b N)C(=O)-C 1～6 alkyl (the -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl in R a and R b each independently represents a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl, or heteroaryl C 1～6 alkyl (e.g., 1 to 10 or 1 to 5) groups are substituted;

[0140] When two hydrogen atoms of the same methylene (-CH 2 -) in formulas (L2-1) to (L2-6) are substituted with C 1～6 alkyl, the alkyls may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0141] The -NH- group in formulas (L2-3) to (L2-6) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0142] m2 is an integer from 1 to 9;

[0143] n2 is an integer from 1 to 18;

[0144] m3 is an integer from 1 to 10;

[0145] n3 is an integer from 1 to 10;

[0146] j2 is an integer from 0 to 9)).

[0147] In this specification, in the structural formula of the linking group (-L 1 -, -L 2 -), among the symbols used to represent integers, when the same symbol is used multiple times, they may be the same integer or different integers respectively.

[0148] In this specification, in Formula (I), Formula (II), Formula (II-P) and Formula (CAL-1), the -CO- of the amide bond (-NH-CO- or -CO-NH-) to which (ALG) is bonded in the formula is the carbonyl group of the carboxyl group derived from alginic acid.

[0149] Hereinafter, each group in Formula (I) and Formula (II) in the aforementioned Mode [1] will be specifically described. In the description of the compound, for example, "C" 1～6 " means that the number of carbon atoms is 1 to 6, and unless otherwise specified, it represents the total number of carbon atoms of a straight-chain, branched-chain or cyclic group. For a group containing a chain-like group and a cyclic group, it means "the total number of carbon atoms of the chain and the ring".

[0150] In addition, unless otherwise specified, each group in the lower modes of Mode [1] also has the same definition as each group in Mode [1].

[0151] In this specification, unless otherwise specified, as "C" 6～10 "aryl", groups such as phenyl, 1-naphthyl, 2-naphthyl, indanyl, indenyl or 1,2,3,4-tetrahydronaphthyl can be cited, for example.

[0152] In this specification, unless otherwise specified, as "heterocyclic group", "heteroaryl" and "non-aromatic heterocyclic group" can be cited, for example.

[0153] In this specification, unless otherwise specified, "heteroaryl" means a monocyclic, polycyclic or fused-ring (wherein, when polycyclic or fused-ring, it can be partially hydrogenated) 5- to 14-membered, preferably 5- to 8-membered, more preferably 5- to 7-membered heteroaryl ring containing 1 to 5, preferably 1 to 3 heteroatoms selected from a nitrogen atom, a sulfur atom, and an oxygen atom.

[0154] In this specification, unless otherwise specified, as the aforementioned "heteroaryl", "monocyclic heteroaryl", "fused-ring heteroaryl", "partially hydrogenated fused-ring heteroaryl" and the like can be cited, for example.

[0155] In this specification, unless otherwise specified, as the aforementioned "monocyclic heteroaryl", it can be the monocyclic form of the aforementioned heteroaryl ring, and the number of ring members is 5 to 8, more preferably 5 to 6 ("5- to 6-membered heteroaryl").

[0156] In this specification, unless otherwise specified, "5- to 6-membered heteroaryl" means a 5- to 6-membered heteroaryl ring containing 1 to 4 heteroatoms selected from a nitrogen atom, a sulfur atom and an oxygen atom, and "5- to 6-membered heteroaryl", unless otherwise specified, means a monovalent group formed by removing any hydrogen atom from the heteroaryl ring.

[0157] In this specification, unless otherwise defined, examples of the aforementioned "5- to 6-membered heteroaryl" include groups such as pyrrolyl, furyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, furazanyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, tetrazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 2H-1,2,3-thiadiazinyl, 4H-1,2,4-thiadiazinyl, 6H-1,3,4-thiadiazinyl, pyridazin-3(2H)-one, pyrimidin-2(1H)-one, pyrazin-2(1H)-one, or pyridin-2(1H)-one.

[0158] In this specification, unless otherwise defined, "5-membered heteroaryl" refers to a 5-membered heteroaryl ring containing 1 to 4 heteroatoms selected from nitrogen, sulfur, and oxygen atoms, and "5-membered heteroaryl", unless otherwise defined, refers to a monovalent group formed by removing any hydrogen atom from the heteroaryl ring. Examples include groups such as pyrrolyl, furyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, furazanyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, or tetrazolyl.

[0159] In this specification, unless otherwise defined, "6-membered heteroaryl" refers to a 6-membered heteroaryl ring containing 1 to 4 heteroatoms selected from nitrogen, sulfur, and oxygen atoms, and "6-membered heteroaryl", unless otherwise defined, refers to a monovalent group formed by removing any hydrogen atom from the heteroaryl ring. Examples include groups such as Pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 2H-1,2,3-thiadiazinyl, 4H-1,2,4-thiadiazinyl, 6H-1,3,4-thiadiazinyl, pyridazin-3(2H)-one, pyrimidin-2(1H)-one, pyrazin-2(1H)-one, or pyridin-2(1H)-one.

[0160] In this specification, unless otherwise defined, "5- to 6-membered heteroaryl C 1～6 alkyl" means that the aforementioned "5- to 6-membered heteroaryl" is substituted by the aforementioned "C 1～6Groups obtained by substituting "alkyl", for example, pyrrolylmethyl, furanylmethyl, thienylmethyl, imidazolylmethyl, pyrazolylmethyl, oxazolylmethyl, isoxazolylmethyl, thiazolylmethyl, isothiazolylmethyl, 1,2,3-triazolylmethyl, 1,2,4-triazolylmethyl, 1,2,3-oxadiazolylmethyl, 1,2,4-oxadiazolylmethyl, 1,3,4-oxadiazolylmethyl, furazanylmethyl, 1,2,3-thiadiazolylmethyl, 1,2,4-thiadiazolylmethyl, 1,3,4-thiadiazolylmethyl, tetrazolylmethyl, pyridylmethyl, pyridazinylmethyl, pyrimidinylmethyl, pyrazinylmethyl, 1,2,3-triazinylmethyl, 1,2,4-triazinylmethyl, 1,3,5-triazinylmethyl, 2H-1,2,3-thiadiazinylmethyl, 4H-1,2,4-thiadiazinylmethyl or 6H-1,3,4-thiadiazinylmethyl and the like.

[0161] "Partially hydrogenated fused heterocyclic aryl" means a monovalent group formed by removing any hydrogen atom from a partially hydrogenated fused ring formed by the condensation of "heterocyclic group" and "aryl" or "heterocyclic group" and "heteroaryl". The any hydrogen atom can be either a hydrogen atom of any ring part of "heterocyclic group", "aryl" and "heteroaryl" in the fused ring or a hydrogen atom of the hydrogenated ring part. For example, if it is a partially hydrogenated tetrahydroquinolinyl group of quinoline, 5,6,7,8-tetrahydroquinolinyl group or 1,2,3,4-tetrahydroquinolinyl group and the like can be cited. These groups depend on the position of removing any hydrogen atom. For example, if it is 5,6,7,8-tetrahydroquinolinyl group, -2-yl, -3-yl, -4-yl, -5-yl, -6-yl, -7-yl, -8-yl and the like can be exemplified. If it is 1,2,3,4-tetrahydroquinolinyl group, for example, -1-yl, -2-yl, -3-yl, -4-yl, -5-yl, -6-yl, -7-yl, -8-yl and the like can be exemplified.

[0162] As the "partially hydrogenated fused polycyclic heteroaryl", those having 8 to 12 ring members are preferred. That is, as the "partially hydrogenated 8- to 12-membered fused polycyclic heteroaryl", examples thereof include indolyl, 2,3-dihydrobenzofuranyl, 4,5,6,7-tetrahydrobenzofuranyl, 2,3-dihydrobenz[d]oxazolyl, 2,3-dihydrobenz[d]thiazolyl, 4,5,6,7-tetrahydrobenz[d]oxazolyl, 4,5,6,7-tetrahydrobenz[d]thiazolyl, 4,5,6,7-tetrahydro-1H-benz[d]imidazolyl, benz[d][1,3]dioxazolyl, 2,3-dihydrobenz[b][1,4]dioxinyl, 2,3-dihydrobenz[b][1,4]oxathiinyl, 3,4-dihydro-2H-benz[b][1,4]oxazinyl, 3,4-dihydro-2H-benz[b][1,4]thiazinyl, 1,2,3,4-tetrahydroquinoxalinyl, 3,4-dihydro-2H-benz[b][1,4]dioxepinyl, 2,3,4,5-tetrahydrobenz[b][1,4]dioxocinyl, N-acetyl-2,3-dihydrobenz[d]oxazolyl, N-acetyl-3,4-dihydro-2H-benz[b][1,4]oxazinyl, N-acetyl-2,3,4,5-tetrahydrobenz[b][1,4]oxazepinyl, N-acetyl-3,4,5,6-tetrahydro-2H-benz[b][1,4]oxazolinyl, N-methanesulfonyl-2,3-dihydrobenz[d]oxazolyl, N-methanesulfonyl-3,4-dihydro-2H-benz[b][1,4]oxazinyl, N-methanesulfonyl-2,3,4,5-tetrahydrobenz[b][1,4]oxazepinyl, N-methanesulfonyl-3,4,5,6-tetrahydro-2H-benz[b][1,4]oxazolinyl, and the like.

[0163] In the present specification, unless otherwise defined, the "non-aromatic heterocyclic group" means a "3- to 14-membered saturated or unsaturated non-aromatic heterocyclic group".

[0164] In the present specification, unless otherwise defined, the "3- to 14-membered saturated or unsaturated non-aromatic heterocyclic group" means a monovalent group formed by removing any hydrogen atom from a 3- to 14-membered saturated or unsaturated heterocyclic ring containing 1 to 4 heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom.

[0165] In this specification, unless otherwise defined, examples of the "non-aromatic heterocyclic group" include groups such as aziridinyl, azetidinyl, oxolanyl, thiopyranyl, oxetanyl, thietanyl, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, thiolanyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, piperidinyl, dihydropyranyl, tetrahydropyranyl (2-tetrahydro-2H-pyranyl, 3-tetrahydro-2H-pyranyl, 4-tetrahydro-2H-pyranyl (4-tetrahydro-2H-pyran-4-yl)), tetrahydrothiopyranyl, piperazinyl, dioxolanyl, oxazolidinyl, isoxazolinyl, 1,3-oxazolidinyl, isoxazolidinyl, thiazolinyl, isothiazolinyl, 1,3-thiazolinyl, isothiazolinyl, oxadiazolinyl, 1,3,4-oxadiazolidinyl, morpholinyl, thiomorpholinyl, quinuclidinyl, azepanyl, diazepinyl or oxepanyl.

[0166] In this specification, unless otherwise defined, examples of the "halogen atom" include a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

[0167] In this specification, unless otherwise defined, "halogenated C" 1～6 In "alkyl", etc., "halogenated" means having several, preferably 1 to 5, of the aforementioned "halogen atoms" as substituents.

[0168] In this specification, unless otherwise defined, examples of "C" 1～6 alkyl include groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl or hexyl.

[0169] In this specification, unless otherwise defined, "halogenated C" 1～6 alkyl means a group obtained by arbitrarily substituting several, preferably 1 to 5, halogen atoms for the aforementioned "C" 1～6 alkyl, and examples thereof include fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 1,1,2,2-tetrafluoroethyl or pentafluoroethyl.

[0170] In this specification, unless otherwise defined, "C" 1～6 alkoxy represents an alkoxy group obtained by bonding the aforementioned "C" 1～6 alkyl to an oxygen atom, and examples thereof include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy or hexyloxy.

[0171] In this specification, unless otherwise defined, "-NR" a R b group means that two hydrogen atoms on the nitrogen atom of "amino" are replaced by -R a 、 -Rb Substituted group.

[0172] In this specification, unless otherwise defined, R a and R b each independently represents a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl, and examples of such groups include an amino group, N-methylamino group, N-ethylamino group, N,N-dimethylamino group, N-acetylamino group, N-methanesulfonylamino group, or N-acetyl-N-methylamino group.

[0173] In this specification, unless otherwise defined, "C 2～7 alkanoyl" refers to "C 1～6 alkyl" bonded to a carbonyl group to form "C 1～6 alkylcarbonyl", and examples of such groups include an acetyl group, propionyl group, butyryl group, isobutyryl group, valeryl group, isovaleryl group, pivaloyl group, hexanoyl group, heptanoyl group, cyclopropylcarbonyl group, cyclobutylcarbonyl group, cyclopentylcarbonyl group, cyclohexylcarbonyl group, cyclopropylmethylcarbonyl group, or 2-methylcyclopropylcarbonyl group.

[0174] In this specification, unless otherwise defined, "C 1～6 alkylsulfonyl" refers to a group obtained by substituting the "sulfonyl group: -SO 2 -" for the aforementioned "C 1～6 alkyl", and examples of such groups include a methylsulfonyl group, ethylsulfonyl group, propylsulfonyl group, or isopropylsulfonyl group.

[0175] In this specification, unless otherwise defined, "cyclic ether" refers to an ether having a structure in which a carbon atom of a cyclic hydrocarbon (e.g., a cyclic hydrocarbon (C 3～8 cycloalkyl ring) having 3 to 8 carbon atoms in a monocyclic or polycyclic saturated hydrocarbon ring group such as cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, or cyclooctane) is replaced by an oxygen atom, and examples of such cyclic ethers include propylene oxide, oxetane, tetrahydrofuran, tetrahydropyran, 1,3-dioxolane, 1,3-dioxane, 1,4-dioxane, 1,3-dioxepane, 1,4-dioxepane, 1,4-dioxane, or 1,5-dioxane.

[0176] In this specification, unless otherwise defined, "3-N-(C 2～7 alkanoyl)oxazolidine ring" refers to a ring in which a hydrogen atom of the NH group of an oxazolidine ring is replaced by the aforementioned "C 2～7 alkanoyl", and examples of such rings include a 3-N-acetyl-oxazolidine ring, 3-N-ethylcarbonyl-oxazolidine ring, etc.

[0177] In this specification, unless otherwise defined, "4-N-(C2～7 The "alkanoyl)morpholine ring" refers to a ring in which the hydrogen atom of the NH group of the morpholine ring is substituted with the aforementioned "C 2～7 alkanoyl", and examples of such rings include a 4-N-acetyl-morpholine ring, a 4-N-ethylcarbonyl-morpholine ring, etc.

[0178] In this specification, unless otherwise defined, "4-N-(C 2～7 alkanoyl)-1,4-oxazepane ring" refers to a ring in which the hydrogen atom of the NH group of the 1,4-oxazepane ring is substituted with the aforementioned "C 2～7 alkanoyl", and examples of such rings include a 4-N-acetyl-1,4-oxazepane ring, a 4-N-ethylcarbonyl-1,4-oxazepane ring, etc.

[0179] In this specification, unless otherwise defined, "3-N-(C 1～6 alkylsulfonyl)oxazolidine ring" refers to a ring in which the hydrogen atom of the NH group of the oxazolidine ring is substituted with the aforementioned "C 1～6 alkylsulfonyl", and examples of such rings include a 3-N-methanesulfonyl-oxazolidine ring, a 3-N-ethylsulfonyl-oxazolidine ring, etc.

[0180] In this specification, unless otherwise defined, "4-N-(C 1～6 alkylsulfonyl)morpholine ring" refers to a ring in which the hydrogen atom of the NH group of the morpholine ring is substituted with the aforementioned "C 1～6 alkylsulfonyl", and examples of such rings include a 4-N-methanesulfonyl-morpholine ring, a 4-N-ethylsulfonyl-morpholine ring, etc.

[0181] In this specification, unless otherwise defined, "4-N-(C 2～7 alkanoyl)-1,4-oxazepane ring" refers to a ring in which the hydrogen atom of the NH group of the 1,4-oxazepane ring is substituted with the aforementioned "C 1～6 alkylsulfonyl", and examples of such rings include a 4-N-methanesulfonyl-1,4-oxazepane ring, a 4-N-ethylsulfonyl-1,4-oxazepane ring, etc.

[0182] In this specification, unless otherwise defined, "hydroxy C 1～6 alkyl" refers to a group in which any hydrogen atom of the aforementioned "C 1～6 alkyl" is optionally substituted with 1 to 5 hydroxy groups, and examples of such groups include a hydroxymethyl group, a 2-hydroxyethyl group, a 3-hydroxypropyl group, or a 2,2-dimethyl-2-hydroxyethyl group (=2-hydroxy-2-methylpropyl), etc.

[0183] In this specification, unless otherwise defined, "mercapto C 1～6 alkyl" refers to a group in which the aforementioned "C 1～6The group in which "alkyl" is optionally substituted by several, preferably 1 to 5, mercapto groups (-SH groups) includes, for example, groups such as mercaptomethyl, 2-mercaptoethyl, or 3-mercaptopropyl.

[0184] In this specification, unless otherwise defined, "C 1～6 alkylthio C 1～6 alkyl" means that the hydrogen atom of the mercapto group (-SH group) of the aforementioned "mercapto C 1～6 alkyl" is substituted by "C 1～6 alkyl", and examples of such groups include methylthiomethyl, methylthioethyl, ethylthiomethyl, or ethylthioethyl.

[0185] In this specification, unless otherwise defined, "-COO(C 1～6 alkyl)" means that the hydrogen atom of the aforementioned "carboxyl group" is substituted by the aforementioned C 1～6 alkyl, and examples of such groups include carboxymethyl, carboxyethyl, carboxypropyl, etc.

[0186] In this specification, unless otherwise defined, "(R a R b N)-C 1～6 alkyl" means that any hydrogen atom of the aforementioned "C 1～6 alkyl" is substituted by "-NR a R b group" (in this specification, R a and R b each independently represents a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl). Examples of such groups include aminomethyl, aminoethyl, aminopropyl, aminobutyl, N-methylaminomethyl, N-acetylaminomethyl, or N-methanesulfonylaminomethyl.

[0187] In this specification, unless otherwise defined, "(R a R b N)C(=O)-C 1～6 alkyl" means that any hydrogen atom of the aforementioned "C 1～6 alkyl" is substituted by "(R a R b N)C(=O)-group" (in this specification, R a and R b each independently represents a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6Groups in the alkylsulfonyl group.) Examples of the optionally substituted group include groups such as aminocarbonylmethyl, aminocarbonylethyl, N-methylaminocarbonylmethyl, N-acetylaminocarbonylethyl, or N-methanesulfonylaminocarbonylethyl.

[0188] In the present specification, unless otherwise defined, "guanidyl C 1～6 alkyl" means a group in which any hydrogen atom of "C 1～6 alkyl" is substituted with "guanidyl (-NH-C(=NH)-NH 2 )". Examples of such groups include guanidylmethyl, guanidylethyl, or guanidylpropyl.

[0189] In the present specification, unless otherwise defined, "C 7～16 aralkyl" means a group in which any hydrogen atom of the aforementioned "C 1～6 alkyl" is substituted with the aforementioned "C 6～10 aryl". Examples of such groups include benzyl, phenethyl, diphenylmethyl, triphenylmethyl, biphenylmethyl, naphthylmethyl, indanylmethyl, or 1,2,3,4-tetrahydronaphthalen-1-ylmethyl.

[0190] In the present specification, unless otherwise defined, "hydroxy C 6～10 aryl C 1～6 alkyl" means a group in which the hydrogen atom of the "C 7～16 aryl" in the aforementioned "C 6～10 aralkyl" is optionally substituted with several, preferably 1 to 5, hydroxy groups. Examples of such groups include 2-hydroxybenzyl, 3-hydroxybenzyl, or 4-hydroxybenzyl.

[0191] In the present specification, unless otherwise defined, "heteroaryl C 1～6 alkyl" means a group in which any hydrogen atom of the aforementioned "heteroaryl" is substituted with the aforementioned "C 1～6 alkyl". Examples of such groups include 2-pyridylmethyl, 4-imidazolylmethyl, or 3-indolylmethyl.

[0192] In the present specification, unless otherwise defined, "non-aromatic heterocycle" means a "3- to 14-membered saturated or unsaturated non-aromatic heterocycle".

[0193] In the present specification, unless otherwise defined, "3- to 14-membered saturated or unsaturated non-aromatic heterocycle" means a 3- to 14-membered saturated or unsaturated heterocycle containing 1 to 4 heteroatoms selected from an oxygen atom, a sulfur atom, and a nitrogen atom.

[0194] In this specification, unless otherwise defined, examples of the "non-aromatic heterocycle" include rings such as aziridine, azetidine, pyrrolidine, pyrazolidine, oxazolidine, thiazolidine, isoxazolidine, isothiazolidine, imidazolidine, piperidine, piperazine, morpholine, thiomorpholine, oxazepane, diazepane, thiazepane, oxadiazole, diazomethane, thiazole, or oxazine.

[0195] In this specification, unless otherwise defined, "C 3～8 cycloalkyl ring" refers to a cyclic saturated hydrocarbon ring having 3 to 8 carbon atoms (including monocyclic or polycyclic), and examples of the ring include cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, or cyclooctane.

[0196] [1-1] In the formula (I) of the foregoing mode [1], -L 1 - is preferably a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0197] [Chemical 19]

[0198]

[0199] (In the formulas (L1-1) to (L1-3), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K, or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 (In the foregoing -NR a R b group, (R a R b N)-C 1～6 alkyl, or (R a R b N)C(=O)-C 1～6 alkyl, R a and R b are each independently selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C1～6 groups in alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 substituted by a plurality of groups in alkyl (for example, 1 to 10 or 1 to 5);

[0200] Two hydrogen atoms of the same methylene (-CH 2 -) in the formulas (L1-1) to (L1-3) are substituted with C 1～6 When the alkyl is an alkyl, the alkyls may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0201] The -NH- group in the formula (L1-3) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0202] n is an integer from 1 to 18;

[0203] m is an integer from 1 to 9;

[0204] j is an integer from 0 to 9);

[0205] More preferably -L 1 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0206] [Chemical formula 20]

[0207]

[0208] (The hydrogen atom of the methylene (-CH 2 -) in the formulas (L1-1) to (L1-3) may be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (the aforementioned -NR a R b group, (Ra R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 R in alkyl a and R b each independently is a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl, and is substituted with a plurality of groups (for example, 1 to 5 groups) therein;

[0209] n is an integer from 1 to 9;

[0210] m is an integer from 1 to 5;

[0211] j is an integer from 0 to 5);

[0212] More preferably, -L 1 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0213] [Chemical formula 21]

[0214]

[0215] (In formulas (L1-1) to (L1-3), the hydrogen atom of methylene (-CH 2 -) can be substituted with a plurality of groups (for example, 1 to 3 groups) selected from a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group or C 7～16 aralkyl);

[0216] n is an integer from 1 to 3;

[0217] m is an integer from 1 to 3;

[0218] j is an integer from 0 to 2);

[0219] Particularly preferably, -L 1 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0220] [Chemical formula 22]

[0221]

[0222] Most preferred - L 1 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0223] [Chemical formula 23]

[0224]

[0225] [1 - 2] In formula (II) of the foregoing method [1], - L 2 - is preferably a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0226] [Chemical formula 24]

[0227]

[0228] (In formulas (L2 - 3), (L2 - 5) and (L2 - 6), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (in the foregoing -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, R a and R b are each independently a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16Arylalkyl, hydroxy C 6～10 Aryl C 1～6 Alkyl or heteroaryl C 1～6 Substituted with multiple groups in the alkyl (e.g., 1 to 10 or 1 to 5);

[0229] In formula (L2-3), formula (L2-5) and formula (L2-6), two hydrogen atoms of the same methylene group (-CH 2 -) are substituted with C 1～6 When the alkyl is alkyl, the alkyls may be bonded to each other to form C 3～8 Cycloalkyl ring;

[0230] In formula (L2-3), formula (L2-5) and formula (L2-6), the -NH- group may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0231] n2 is an integer from 1 to 18;

[0232] m3 is an integer from 1 to 10;

[0233] n3 is an integer from 1 to 10;

[0234] j2 is an integer from 0 to 9);

[0235] More preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0236] [Chemical formula 25]

[0237]

[0238] (In formula (L2-3), formula (L2-5) and formula (L2-6), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (the aforementioned -NR a Rb group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 R in alkyl a and R b each independently is a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl (for example, 1 to 10 or 1 to 5) groups are substituted;

[0239] n2 is an integer from 1 to 9;

[0240] m3 is an integer from 1 to 6;

[0241] n3 is an integer from 1 to 6;

[0242] j2 is an integer from 0 to 6);

[0243] More preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0244] [Chemical formula 26]

[0245]

[0246] (In formula (L2-5-1) and formula (L2-6-1), the R A groups each independently are a group selected from a hydrogen atom, a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group or C 7～16 aralkyl);

[0247] n2 is an integer from 1 to 5;

[0248] m3 is an integer from 1 to 3;

[0249] n3 is an integer from 1 to 4;

[0250] j2 is an integer from 0 to 3);

[0251] Most preferably -L2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0252] [Chemical formula 27]

[0253]

[0254] Most preferably -L 2 - is the following partial structural formula [in each formula, the outside of the dotted lines at both ends is not included]:

[0255] [Chemical formula 28]

[0256]

[0257] [1 - 2a] In formula (II) of the aforementioned method [1], -L 2 - is preferably a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0258] [Chemical formula 29]

[0259]

[0260] (In formula (L2 - 3), formula (L2 - 4), formula (L2 - 5) and formula (L2 - 6), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (in the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, R a and R b each independently is selected from a hydrogen atom, C1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl group), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl, or heteroaryl C 1～6 alkyl groups (e.g., 1 to 10 or 1 to 5) are substituted;

[0261] In formula (L2-3), formula (L2-4), formula (L2-5), and formula (L2-6), two hydrogen atoms of the same methylene group (-CH 2 -) are substituted with C 1～6 alkyl, and the alkyls may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0262] In formula (L2-3), formula (L2-4), formula (L2-5), and formula (L2-6), the -NH- group may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0263] n2 is an integer from 1 to 18;

[0264] m3 is an integer from 1 to 10;

[0265] n3 is an integer from 1 to 10;

[0266] j2 is an integer from 0 to 9);

[0267] More preferably, -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0268] [Chemical formula 30]

[0269]

[0270] (In formula (L2-4) and formula (L2-6), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K, or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R bN)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 R in alkyl a and R b each independently is a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl, or heteroaryl C 1～6 alkyl (e.g., 1 to 10 or 1 to 5) groups are substituted;

[0271] n2 is an integer from 1 to 9;

[0272] m3 is an integer from 1 to 6;

[0273] n3 is an integer from 1 to 6;

[0274] j2 is an integer from 0 to 6);

[0275] More preferably, -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0276] [Chemical Formula 31]

[0277]

[0278] (In formula (L2-4-1) and formula (L2-6-1), R A groups each independently is a group selected from a hydrogen atom, a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K, or 1 / 2Ca), -COO(C 1～6 alkyl) group, or C 7～16 aralkyl);

[0279] n2 is an integer from 1 to 5;

[0280] m3 is an integer from 1 to 3;

[0281] n3 is an integer from 1 to 4;

[0282] j2 is an integer from 0 to 3);

[0283] Particularly preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0284] [Chemical formula 32]

[0285]

[0286] Most preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0287] [Chemical formula 33]

[0288]

[0289] [1-2b] In formula (II) of the aforementioned method [1], -L 2 - is preferably a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0290] [Chemical formula 34]

[0291]

[0292] (In formula (L2-3), formula (L2-4), formula (L2-5) and formula (L2-6), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (the aforementioned -NR a R b group, (R a R b N)-C 1～6alkyl or (R a R b N)C(=O)-C 1～6 R in alkyl a and R b each independently is a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl, or heteroaryl C 1～6 alkyl, and is substituted with a plurality of groups (for example, 1 to 10 or 1 to 5);

[0293] When two hydrogen atoms of the same methylene group (-CH 2 -) in Formula (L2-3), Formula (L2-4), Formula (L2-5), and Formula (L2-6) are substituted with C 1～6 alkyl, the alkyls may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0294] The -NH- group in Formula (L2-3), Formula (L2-4), Formula (L2-5), and Formula (L2-6) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0295] n2 is an integer from 1 to 18;

[0296] m3 is an integer from 1 to 10;

[0297] n3 is an integer from 1 to 10;

[0298] j2 is an integer from 0 to 9);

[0299] More preferably, -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0300] [Chemical Formula 35]

[0301]

[0302] (The hydrogen atom of the methylene group (-CH 2 -) in Formula (L2-3), Formula (L2-4), Formula (L2-5), and Formula (L2-6) may be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6An alkyl group, -COOH group, -COOM group (where M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (in the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, R a and R b each independently represents a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl), a guanidino C 1～6 alkyl, C 7～16 aralkyl, a hydroxy C 6～10 aryl C 1～6 alkyl or a heteroaryl C 1～6 alkyl (for example, 1 to 10 or 1 to 5) groups are substituted;

[0303] In formula (L2-3), formula (L2-4), formula (L2-5) and formula (L2-6), the -NH- group can form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0304] n2 is an integer from 1 to 9;

[0305] m3 is an integer from 1 to 6;

[0306] n3 is an integer from 1 to 6;

[0307] j2 is an integer from 0 to 6);

[0308] More preferably, -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0309] [Chemical formula 36]

[0310]

[0311] (In formula (L2-3-1), formula (L2-4-1), formula (L2-5-1) and formula (L2-6-1), RA Each of the groups is independently a group selected from a hydrogen atom, a halogen atom, a hydroxyl group, a C 1～6 alkyl group, a hydroxy C 1～6 alkyl group, a -COOH group, a -COOM group (where M is Li, Na, K or 1 / 2Ca), a -COO(C 1～6 alkyl) group or a C 7～16 aralkyl group;

[0312] In formula (L2-3-1), formula (L2-4-1), formula (L2-5-1) and formula (L2-6-1), the -NH- group may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0313] n2 is an integer of 1 to 5;

[0314] m3 is an integer of 1 to 3;

[0315] n3 is an integer of 1 to 4;

[0316] j2 is an integer of 0 to 3);

[0317] Particularly preferably, it is a linking group selected from the following

[0318] [Chemical formula 37]

[0319]

[0320] (In formula (L2-4-1), formula (L2-6-1) and formula (L2-6-1-a), each R A group is independently a group selected from a hydrogen atom, a halogen atom, a hydroxyl group, a C 1～6 alkyl group, a hydroxy C 1～6 alkyl group, a -COOH group, a -COOM group (where M is Li, Na, K or 1 / 2Ca), a -COO(C 1～6 alkyl) group or a C 7～16 aralkyl group;

[0321] n2 is an integer of 1 to 5;

[0322] m3 is an integer of 1 to 3;

[0323] n3 is an integer of 1 to 4;

[0324] j2 is an integer of 0 to 3;

[0325] k is an integer of 1 to 4);

[0326] Most preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0327] [Chemical formula 38]

[0328]

[0329] Further most preferably - L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0330] [Chemical formula 39]

[0331]

[0332] [2] The second mode is as follows. An alginic acid derivative represented by the following formula (I):

[0333] [Chemical formula 40]

[0334]

[0335] (In formula (I), (ALG) represents alginic acid; -NHCO- represents an amide bond obtained through an arbitrary carboxyl group of alginic acid; -L 1 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0336] [Chemical formula 41]

[0337]

[0338] (In formulas (L1-1) to (L1-4), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C1～6 R in the alkyl a and R b are each independently a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl, or heteroaryl C 1～6 alkyl, and are substituted with a plurality of groups (for example, 1 to 10 or 1 to 5);

[0339] When two hydrogen atoms of the same methylene (-CH 2 -) in formulas (L1-1) to (L1-4) are substituted with C 1～6 alkyl, the alkyls may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0340] The -NH- group in formulas (L1-3) and (L1-4) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0341] n is an integer from 1 to 18;

[0342] m is an integer from 1 to 9;

[0343] j is an integer from 0 to 9)).

[0344] [2-1] In formula (I) of the aforementioned method [2], -L 1 - is preferably a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0345] [Chemical formula 42]

[0346]

[0347] (The hydrogen atom of the methylene (-CH 2 -) in formulas (L1-1) to (L1-3) may be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R bN)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 R in alkyl a and R b each independently is a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl by multiple groups (for example, 1 to 10 or 1 to 5);

[0348] When two hydrogen atoms of the same methylene (-CH 2 -) in Formula (L1-1) to Formula (L1-3) are substituted with C 1～6 alkyl, the alkyls may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0349] The -NH- group in Formula (L1-3) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0350] n is an integer from 1 to 18;

[0351] m is an integer from 1 to 9;

[0352] j is an integer from 0 to 9);

[0353] More preferably, -L 1 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0354] [Chemical formula 43]

[0355]

[0356] (The hydrogen atom of the methylene (-CH 2 -) in Formula (L1-1) to Formula (L1-3) may be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6Alkyl, mercaptan C 1～6 Alkyl, C 1～6 Alkylthio C 1～6 Alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 Alkyl) group, -NR a R b Group, (R a R b N)-C 1～6 Alkyl, (R a R b N)C(=O)-C 1～6 Alkyl (the aforementioned -NR a R b Group, (R a R b N)-C 1～6 Alkyl or (R a R b N)C(=O)-C 1～6 In alkyl, R a And R b Each independently is a group selected from a hydrogen atom, C 1～6 Alkyl, C 2～7 Alkanoyl or C 1～6 Alkylsulfonyl), guanidyl C 1～6 Alkyl, C 7～16 Aralkyl, hydroxy C 6～10 Aryl C 1～6 Alkyl or heteroaryl C 1～6 Alkyl, and is substituted by a plurality of groups (for example, 1 to 5) among them;

[0357] n is an integer from 1 to 9;

[0358] m is an integer from 1 to 5;

[0359] j is an integer from 0 to 5);

[0360] More preferably -L 1 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0361] [Chemical formula 44]

[0362]

[0363] (In formula (L1-1) to formula (L1-3), the hydrogen atom of methylene (-CH 2 -) can be selected from a halogen atom, a hydroxy group, C 1～6 Alkyl, hydroxy C 1～6An alkyl group, -COOH group, -COOM group (where M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group or multiple groups (e.g., 1 to 3) of an aralkyl group are substituted; 7～16

[0364] n is an integer from 1 to 3;

[0365] m is an integer from 1 to 3;

[0366] j is an integer from 0 to 2);

[0367] Particularly preferably, -L 1 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0368] [Chemical formula 45]

[0369]

[0370] Most preferably, -L 1 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0371] [Chemical formula 46]

[0372]

[0373] [3] The third mode is as follows. The alginic acid derivative described in the above [2], wherein, in formula (BR-1):

[0374] [Chemical formula 47]

[0375]

[0376] (In formula (BR-1), -L 1 - is the same as defined in the above mode [1] or [2]) The introduction rate of the group shown is 1% to 30%.

[0377] [4] The fourth mode is as follows. The alginic acid derivative described in the above [2], wherein the weight-average molecular weight of the alginic acid derivative shown in formula (I) measured by gel filtration chromatography is 100,000 Da to 3,000,000 Da.

[0378] [5] The fifth mode is as follows. The alginic acid derivative shown in the following formula (II-P):

[0379] [Chemical formula 48]

[0380]

[0381] ​(In formula (II-P), (ALG) represents alginic acid; -NHCO- represents an amide bond formed from any carboxyl group of alginic acid; P 1 is a hydrogen atom or a protecting group for a mercapto group (-SH group); -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0382] [Chemical formula 49]

[0383]

[0384] (The hydrogen atom of the methylene group (-CH 2 -) in formulas (L2-1) to (L2-6) can be substituted by a group selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (in the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, R a and R b each independently is a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl by multiple groups (for example, 1 to 10 or 1 to 5) substitution;

[0385] In formulas (L2-1) to (L2-6), two hydrogen atoms of the same methylene group (-CH 2 -) are substituted by C1～6 When the alkyl group is an alkyl group, the alkyl groups can be bonded to each other to form a C 3～8 cycloalkyl ring;

[0386] The -NH- group in the formula (L2-3) to the formula (L2-6) can form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0387] m2 is an integer from 1 to 9;

[0388] n2 is an integer from 1 to 18;

[0389] m3 is an integer from 1 to 10;

[0390] n3 is an integer from 1 to 10;

[0391] j2 is an integer from 0 to 9)).

[0392] [5-1] In the formula (II-P) of the foregoing method [5], preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0393] [Chemical formula 50]

[0394]

[0395] (In the formula (L2-3), the formula (L2-5) and the formula (L2-6), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (the foregoing -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, the R ina and R b each independently represents a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl, and is substituted with a plurality of groups (for example, 1 to 10 or 1 to 5);

[0396] When two hydrogen atoms of the same methylene group (-CH 2 -) in formula (L2-3), formula (L2-5) and formula (L2-6) are substituted with C 1～6 alkyl, the alkyls may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0397] The -NH- group in formula (L2-3), formula (L2-5) and formula (L2-6) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0398] n2 is an integer from 1 to 18;

[0399] m3 is an integer from 1 to 10;

[0400] n3 is an integer from 1 to 10;

[0401] j2 is an integer from 0 to 9);

[0402] More preferably, -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0403] [Chemical formula 51]

[0404]

[0405] (The hydrogen atom of the methylene group (-CH 2 -) in formula (L2-3), formula (L2-5) and formula (L2-6) may be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R bGroup, (R a R b N)-C 1～6 alkyl group, (R a R b N)C(=O)-C 1～6 alkyl group (the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl group or (R a R b N)C(=O)-C 1～6 in the alkyl group, R a and R b each independently represents a group selected from a hydrogen atom, C 1～6 alkyl group, C 2～7 alkanoyl group or C 1～6 alkylsulfonyl group), guanidino C 1～6 alkyl group, C 7～16 aralkyl group, hydroxy C 6～10 aryl C 1～6 alkyl group or heteroaryl C 1～6 alkyl group (for example, 1 to 10 or 1 to 5) are substituted;

[0406] n2 is an integer from 1 to 9;

[0407] m3 is an integer from 1 to 6;

[0408] n3 is an integer from 1 to 6;

[0409] j2 is an integer from 0 to 6);

[0410] More preferably, -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0411] [Chemical Formula 52]

[0412]

[0413] (In formula (L2-5-1) and formula (L2-6-1), the R A groups each independently represent a group selected from a hydrogen atom, a halogen atom, a hydroxyl group, C 1～6 alkyl group, hydroxy C 1～6 alkyl group, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group or C 7～16 aralkyl group);

[0414] n2 is an integer from 1 to 5;

[0415] m3 is an integer from 1 to 3;

[0416] n3 is an integer from 1 to 4;

[0417] j2 is an integer from 0 to 3);

[0418] Particularly preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0419] [Chemical formula 53]

[0420]

[0421] Most preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0422] [Chemical formula 54]

[0423]

[0424] [5-1a] In formula (II-P) of the aforementioned method [5], -L 2 - is preferably a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0425] [Chemical formula 55]

[0426]

[0427] (In formula (L2-3), formula (L2-4), formula (L2-5) and formula (L2-6), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (the aforementioned -NR a R b group, (R a Rb N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 R in the alkyl a and R b each independently is a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl (for example, 1 to 10 or 1 to 5) of the plurality of groups are substituted;

[0428] When two hydrogen atoms of the same methylene (-CH 2 -) in the formula (L2-3), formula (L2-4), formula (L2-5) and formula (L2-6) are substituted with C 1～6 alkyl, the alkyls may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0429] The -NH- group in the formula (L2-3), formula (L2-4), formula (L2-5) and formula (L2-6) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0430] n2 is an integer of 1 to 18;

[0431] m3 is an integer of 1 to 10;

[0432] n3 is an integer of 1 to 10;

[0433] j2 is an integer of 0 to 9);

[0434] More preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0435] [Chemical formula 56]

[0436]

[0437] (The hydrogen atom of the methylene (-CH 2 -) in the formula (L2-4) and formula (L2-6) may be selected from an oxo group (=O), a halogen atom, a hydroxy group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C1～6 Alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (in the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, R a and R b each independently represents a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl are each substituted by a plurality of groups (for example, 1 to 10 or 1 to 5);

[0438] n2 is an integer from 1 to 9;

[0439] m3 is an integer from 1 to 6;

[0440] n3 is an integer from 1 to 6;

[0441] j2 is an integer from 0 to 6);

[0442] More preferably, -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0443] [Chemical formula 57]

[0444]

[0445] (In formula (L2-4-1) and formula (L2-6-1), R A groups each independently represent a group selected from a hydrogen atom, a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6An alkyl group, a -COOH group, a -COOM group (where M is Li, Na, K or 1 / 2Ca), a -COO(C 1～6 alkyl) group or a C 7～16 arylalkyl group;

[0446] n2 is an integer from 1 to 5;

[0447] m3 is an integer from 1 to 3;

[0448] n3 is an integer from 1 to 4;

[0449] j2 is an integer from 0 to 3);

[0450] Particularly preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0451] [Chemical formula 58]

[0452]

[0453] Most preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0454] [Chemical formula 59]

[0455]

[0456] [5-1b] In the formula (II-P) of the foregoing method [5], -L 2 - is preferably the following partial structural formula [in each formula, the outside of the dotted lines at both ends is not included]:

[0457] [Chemical formula 60]

[0458]

[0459] (In the methylene (-CH 2 -) in the formula (L2-3), formula (L2-4), formula (L2-5) and formula (L2-6), the hydrogen atom can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, a C 1～6 alkyl group, a hydroxy C 1～6 alkyl group, a mercapto C 1～6 alkyl group, a C 1～6 alkylthio C 1～6 alkyl group, a -COOH group, a -COOM group (where M is Li, Na, K or 1 / 2Ca), a -COO(C 1～6 alkyl) group, a -NR a R b group, (R a R bN)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 R in the alkyl a and R b each independently is a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl, or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl, or heteroaryl C 1～6 alkyl (e.g., 1 to 10 or 1 to 5) groups are substituted;

[0460] When two hydrogen atoms of the same methylene (-CH 2 -) in formula (L2-3), formula (L2-4), formula (L2-5), and formula (L2-6) are substituted with C 1～6 alkyl, the alkyls may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0461] The -NH- group in formula (L2-3), formula (L2-4), formula (L2-5), and formula (L2-6) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0462] n2 is an integer from 1 to 18;

[0463] m3 is an integer from 1 to 10;

[0464] n3 is an integer from 1 to 10;

[0465] j2 is an integer from 0 to 9);

[0466] More preferably, -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0467] [Chemical formula 61]

[0468]

[0469] (In formulas (L2-3), (L2-4), (L2-5) and (L2-6), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (in the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, R a and R b are each independently selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl (for example, 1 to 10 or 1 to 5) groups are substituted;

[0470] In formulas (L2-3), (L2-4), (L2-5) and (L2-6), the -NH- group can form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0471] n2 is an integer from 1 to 9;

[0472] m3 is an integer from 1 to 6;

[0473] n3 is an integer from 1 to 6;

[0474] j2 is an integer from 0 to 6);

[0475] More preferably -L 2- A linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0476] [Chemical formula 62]

[0477]

[0478] (In formula (L2-3-1), formula (L2-4-1), formula (L2-5-1) and formula (L2-6-1), R A groups are each independently selected from a hydrogen atom, a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxyl C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group or C 7～16 aralkyl groups;

[0479] The -NH- group in formula (L2-3-1), formula (L2-4-1), formula (L2-5-1) and formula (L2-6-1) can form a non-aromatic heterocycle together with the substituents bonded to the adjacent carbon atoms;

[0480] n2 is an integer from 1 to 5;

[0481] m3 is an integer from 1 to 3;

[0482] n3 is an integer from 1 to 4;

[0483] j2 is an integer from 0 to 3);

[0484] Particularly preferred is a linking group selected from the following,

[0485] [Chemical formula 63]

[0486]

[0487] (In formula (L2-4-1), formula (L2-6-1) and formula (L2-6-1-a), R A groups are each independently selected from a hydrogen atom, a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxyl C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group or C 7～16 aralkyl groups;

[0488] n2 is an integer from 1 to 5;

[0489] m3 is an integer from 1 to 3;

[0490] n3 is an integer from 1 to 4;

[0491] j2 is an integer from 0 to 3;

[0492] k is an integer from 1 to 4);

[0493] Most preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0494] [Chemical formula 64]

[0495]

[0496] Even more preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0497] [Chemical formula 65]

[0498]

[0499] [5-2] In the formula (II-P) of the above-mentioned method [5], preferably P 1 is a hydrogen atom, an acetyl group or a benzoyl group.

[0500] [5-2a] In the formula (II-P) of the above-mentioned method [5], P 1 is preferably a hydrogen atom, an acetyl group or a benzoyl group, and more preferably a hydrogen atom or a benzoyl group.

[0501] [6-1] The 6-1 method is as follows. The alginic acid derivative represented by the formula (II) described in the above-mentioned method [1], wherein the following formula (BR-2):

[0502] [Chemical formula 66]

[0503]

[0504] (In the formula (BR-2), -L 2 - has the same definition as in the above-mentioned method [1] or [5]) and the introduction rate of the reactive group is 1% to 30%.

[0505] [6-2] The 6-2 method is as follows. The alginic acid derivative represented by the formula (II-P) described in the above-mentioned [5], wherein the following formula (BR-2-P):

[0506] [Chemical formula 67]

[0507]

[0508] (In the formula (BR-2-P), P 1 , -L 2- The introduction rate of the reactive group shown in the same definition as in the aforementioned method [1] or [5] is 1% to 30%.

[0509] [7-1] The 7-1 method is as follows. The alginic acid derivative represented by formula (II) described in the aforementioned method [1], wherein the weight-average molecular weight of the alginic acid derivative measured by gel filtration chromatography is 100,000 Da to 3,000,000 Da.

[0510] [7-2] The 7-2 method is as follows. The alginic acid derivative represented by formula (II-P) described in the aforementioned method [5], wherein the weight-average molecular weight of the alginic acid derivative measured by gel filtration chromatography is 100,000 Da to 3,000,000 Da.

[0511] [8] The 8th method is as follows. The crosslinked alginic acid described in the aforementioned [1], in the crosslinked alginic acid of the aforementioned method [1], the chemical crosslinking is a structure of the following formula (LK-1):

[0512] [Chemical formula 68]

[0513]

[0514] [In formula (LK-1), the -CONH- and -NHCO- at both ends represent amide bonds obtained through any carboxyl group of alginic acid; -L 1 -, and -L 2 - is the same as the definition in the aforementioned method [1].

[0515] [8-1] The 8-1 method is as follows. In the aforementioned method [8], preferably, more preferably, further preferably, particularly preferably, or most preferably, the linking group -L 1 - is the same as the definition in the aforementioned method [2-1].

[0516] [8-2] The 8-2 method is as follows. In the aforementioned method [8], preferably, more preferably, further preferably, particularly preferably, or most preferably, the linking group -L 2 - is the same as the definition in the aforementioned method [5-1].

[0517] [8-3] The 8-3 method is as follows. In the aforementioned method [8], preferably, more preferably, further preferably, particularly preferably, or most preferably, the linking group -L 2 - is the same as the definition in the aforementioned method [5-1a].

[0518] [8-4] The 8-4 method is as follows. In the aforementioned method [8], preferably, more preferably, further preferably, particularly preferably, most preferably, or further most preferably, the linking group -L 2 - is the same as the definition in the aforementioned method [5-1b].

[0519] [8a] The 8a mode is as described below. Crosslinked alginic acid shown in the following formula (CAL-1):

[0520] [Chemical Formula 69]

[0521]

[0522] [In formula (CAL-1), -CONH- and -NHCO- represent amide bonds obtained through any carboxyl group of alginic acid; (ALG), -L 1 -, and -L 2 - have the same definitions as in the aforementioned mode [1]; the two (ALG) in formula (CAL-1) are alginic acids derived from the alginic acid derivative of formula (I) or the alginic acid derivative of formula (II).

[0523] [8a-1] The 8a-1 mode is as described below. Crosslinked alginic acid shown in the following formula (CAL-1) obtained by using the crosslinking reaction described in the aforementioned mode [1]:

[0524] [Chemical Formula 70]

[0525]

[0526] [In formula (CAL-1), -CONH- and -NHCO- represent amide bonds obtained through any carboxyl group of alginic acid; (ALG), -L 1 -, and -L 2 - have the same definitions as in the aforementioned mode [1]; the two (ALG) in formula (CAL-1) are alginic acids derived from the alginic acid derivative of formula (I) or the alginic acid derivative of formula (II).

[0527] [8a-2] The 8a-2 mode is as described below. In the aforementioned mode [8a] or [8a-1], the preferred, more preferred, further preferred, particularly preferred, or most preferred linking group -L 1 - has the same definition as in the aforementioned mode [2-1].

[0528] [8a-3] The 8a-3 mode is as described below. In the aforementioned mode [8a] or [8a-1], the preferred, more preferred, further preferred, particularly preferred, or most preferred linking group -L 2 - has the same definition as in the aforementioned mode [5-1].

[0529] [8a-4] The 8a-4 mode is as described below. In the aforementioned mode [8a] or [8a-1], the preferred, more preferred, further preferred, particularly preferred, or most preferred linking group -L 2 - has the same definition as in the aforementioned mode [5-1a].

[0530] [8a-5] The 8a-5 mode is as described below. In the foregoing mode [8a] or [8a-1], preferably, more preferably, further preferably, particularly preferably, most preferably or further most preferably, the linking group -L 2 - is the same as defined in the foregoing mode [5-1b].

[0531] [9] The 9th mode is as described below. A method for preparing crosslinked alginic acid as described in the foregoing mode [1], which comprises carrying out a crosslinking reaction of a solution of an alginic acid derivative represented by formula (I) in a solution of an alginic acid derivative represented by formula (II).

[0532]

[10] The 10th mode is as described below. A method for preparing crosslinked alginic acid as described in the foregoing mode [1], which comprises carrying out a crosslinking reaction of a solution of an alginic acid derivative represented by formula (II) in a solution of an alginic acid derivative represented by formula (I).

[0533]

[11] The 11th mode is as described below. A method for preparing crosslinked alginic acid as described in the foregoing [1], wherein the chemical crosslinking formed by carrying out a Michael addition reaction using an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II) has the structure of the following formula (LK-1):

[0534] [Chemical formula 71]

[0535]

[0536] [In formula (LK-1), the -CONH- and -NHCO- at both ends represent amide bonds obtained through any carboxyl group of alginic acid; -L 1 -, and -L 2 - are the same as defined in the foregoing mode [1].]

[0537] [11-1] The 11-1 mode is as described below. In the foregoing mode

[11] , preferably, more preferably, further preferably, particularly preferably or most preferably, the linking group -L 1 - is the same as defined in the foregoing mode [2-1].

[0538] [11-2] The 11-2 mode is as described below. In the foregoing mode

[11] , preferably, more preferably, further preferably, particularly preferably or most preferably, the linking group -L 2 - is the same as defined in the foregoing mode [5-1].

[0539] [11-3] The 11-3 mode is as described below. In the foregoing mode

[11] , preferably, more preferably, further preferably, particularly preferably or most preferably, the linking group -L 2 - is the same as defined in the foregoing mode [5-1a].

[0540] [11-4] The 11-4 mode is as described below. In the aforementioned mode

[11] , the linking group -L that is preferably, more preferably, further preferably, particularly preferably, most preferably, or further most preferably 2 - is the same as defined in the aforementioned mode [5-1b].

[0541]

[12] The 12th mode is as described below. The crosslinked alginic acid structure is obtained by subjecting the gel obtained by dropping a solution of the alginic acid derivative represented by formula (I) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of the alginic acid derivative represented by formula (II). As the crosslinking, it includes ionic crosslinking partially formed by the divalent metal ion and chemical crosslinking formed by a Michael addition reaction.

[0542]

[13] The 13th mode is as described below. The crosslinked alginic acid structure is obtained by subjecting the gel obtained by dropping a solution of the alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of the alginic acid derivative represented by formula (I). As the crosslinking, it includes ionic crosslinking partially formed by the divalent metal ion and chemical crosslinking formed by a Michael addition reaction.

[0543]

[14] The 14th mode is as described below. The crosslinked alginic acid structure is obtained by dropping a solution of a composition containing the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion. As the crosslinking, it includes ionic crosslinking partially formed by the divalent metal ion and chemical crosslinking formed by a Michael addition reaction.

[0544]

[15] The 15th mode is as described below. In any one of the aforementioned

[12] to

[14] , the crosslinked alginic acid structure, wherein the chemical crosslinking formed by the Michael addition reaction using the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) has the structure of the following formula (LK-1):

[0545] [Chemical formula 72]

[0546]

[0547] [In formula (LK-1), the -CONH- and -NHCO- at both ends represent amide bonds formed through any carboxyl group of alginic acid; -L 1 -, and -L 2 - is the same as defined in the aforementioned mode [1].]

[0548] [15-1] The 15-1 mode is as described below. In the aforementioned mode

[15] , the linking group -L that is preferably, more preferably, further preferably, particularly preferably, or most preferably 1- The same as the definition in the foregoing method [2-1].

[0549] [15-2] The 15-2 method is as follows. In the foregoing method

[15] , the preferred, more preferred, further preferred, particularly preferred or most preferred linking group -L 2 - The same as the definition in the foregoing method [5-1].

[0550] [15-3] The 15-3 method is as follows. In the foregoing method

[15] , the preferred, more preferred, further preferred, particularly preferred or most preferred linking group -L 2 - The same as the definition in the foregoing method [5-1a].

[0551] [15-4] The 15-4 method is as follows. In the foregoing method

[15] , the preferred, more preferred, further preferred, particularly preferred, most preferred or further most preferred linking group -L 2 - The same as the definition in the foregoing method [5-1b].

[0552]

[16] The 16th method is as follows. The crosslinked alginic acid structure described in any one of the foregoing methods

[12] to

[15] is a fibrous structure, fiber, bead, gel or nearly spherical gel.

[0553]

[17] The 17th method is as follows. A medical material comprising the crosslinked alginic acid structure described in any one of the foregoing methods

[12] to

[16] .

[0554]

[18] The 18th method is as follows. The medical material described in the foregoing method

[17] is a fibrous structure, fiber, bead, gel or nearly spherical gel.

[0555]

[19] The 19th method is as follows. A composition containing an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II).

[0556] [19-1] The 19-1 method is as follows. A composition containing an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II-P).

[0557] [19-2] The 19-2 method is as follows. A composition containing an alginic acid derivative selected from an alginic acid derivative represented by formula (I), an alginic acid derivative represented by formula (II) or an alginic acid derivative represented by formula (II-P).

[0558]

[20] The 20th method is as follows. A method for preparing a crosslinked alginic acid structure, which includes dropping a solution of a composition containing an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion.

[0559]

[21] The 21st mode is as described below. A method for preparing a crosslinked alginic acid structure, comprising: subjecting a gel obtained by dropping a solution of an alginic acid derivative represented by formula (I) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of an alginic acid derivative represented by formula (II), thereby obtaining a crosslinked alginic acid structure containing an ionic crosslink partially formed by a divalent metal ion and a chemical crosslink formed by a Michael addition reaction as crosslinks.

[0560]

[22] The 22nd mode is as described below. A method for preparing a crosslinked alginic acid structure, comprising: subjecting a gel obtained by dropping a solution of an alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of an alginic acid derivative represented by formula (I), thereby obtaining a crosslinked alginic acid structure containing an ionic crosslink partially formed by a divalent metal ion and a chemical crosslink formed by a Michael addition reaction as crosslinks.

[0561]

[23] The 23rd mode is as described below. The method for preparing a crosslinked alginic acid structure according to any one of the foregoing modes

[20] to

[22] , wherein the chemical crosslink formed by a Michael addition reaction using the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) has the structure of the following formula (LK-1):

[0562] [Chemical formula 73]

[0563]

[0564] [In formula (LK-1), the -CONH- and -NHCO- at both ends represent amide bonds formed through any carboxyl group of alginic acid; -L 1 -, and -L 2 - are the same as defined in the foregoing mode [1]].

[0565] [23-1] The 23-1st mode is as described below. In the foregoing mode

[23] , the preferred, more preferred, further preferred, particularly preferred, or most preferred linking group -L 1 - is the same as defined in the foregoing mode [2-1].

[0566] [23-2] The 23-2nd mode is as described below. In the foregoing mode

[23] , the preferred, more preferred, further preferred, particularly preferred, or most preferred linking group -L 2 - is the same as defined in the foregoing mode [5-1].

[0567] [23-3] The 23-3rd mode is as described below. In the foregoing mode

[23] , the preferred, more preferred, further preferred, particularly preferred, or most preferred linking group -L 2- The same as the definition in the foregoing method [5-1a].

[0568] [23-4] The 23-4 method is as described below. In the foregoing method

[23] , the linking group -L that is preferably, more preferably, further preferably, particularly preferably, most preferably or further most preferably 2 - The same as the definition in the foregoing method [5-1b].

[0569]

[24] The 24th method is as described below. The crosslinked alginic acid structure is obtained by subjecting the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) to ionic crosslinking using a divalent metal ion and chemical crosslinking using a Michael addition reaction, and has the property of retaining the content.

[0570]

[25] The 25th method is as described below. The crosslinked alginic acid described in the foregoing method [1] or [8] or the crosslinked alginic acid structure described in any one of the foregoing methods

[12] to

[16] and

[24] has biocompatibility.

[0571] [26a] The 26a method is as described below. The alginic acid derivative represented by formula (I) described in the foregoing method [2] and the alginic acid derivative represented by formula (II-P) described in the foregoing method [5] have biocompatibility.

[0572]

[27] The 27th method is as described below. The amino compound represented by the following formula (AM-2) or a pharmaceutically acceptable salt thereof:

[0573] [Chemical formula 74]

[0574]

[0575] (In formula (AM-2), P 1 , -L 2 - is the same as the definition in the foregoing method [5]).

[0576] [27-1] In formula (AM-2) of the foregoing method

[27] , -L 2 - is preferably a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0577] [Chemical formula 75]

[0578]

[0579] (In formulas (L2-3), (L2-4), (L2-5) and (L2-6), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6Alkyl, mercapto C 1～6 Alkyl, C 1～6 Alkylthio C 1～6 Alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -NR a R b group, (R a R b N)-C 1～6 Alkyl, (R a R b N)C(=O)-C 1～6 Alkyl (the aforementioned -NR a R b group, (R a R b N)-C 1～6 Alkyl or (R a R b N)C(=O)-C 1～6 R in alkyl a and R b each independently is a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl are substituted by a plurality of groups (for example, 1 to 10 or 1 to 5);

[0580] When two hydrogen atoms of the same methylene (-CH 2 -) in formula (L2-3), formula (L2-4), formula (L2-5) and formula (L2-6) are substituted with C 1～6 alkyl, the alkyls may be bonded to each other to form a C 3～8 cycloalkyl ring;

[0581] The -NH- group in formula (L2-3), formula (L2-4), formula (L2-5) and formula (L2-6) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0582] n2 is an integer from 1 to 18;

[0583] m3 is an integer from 1 to 10;

[0584] n3 is an integer from 1 to 10;

[0585] j2 is an integer from 0 to 9);

[0586] More preferably -L 2- A linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0587] [Chemical formula 76]

[0588]

[0589] (In formula (L2-3), formula (L2-4), formula (L2-5) and formula (L2-6), the hydrogen atom of the methylene group (-CH 2 -) can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (in the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, R a and R b are each independently selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl groups), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl by multiple groups (for example, 1 to 10 or 1 to 5); the -NH- group in formula (L2-3), formula (L2-4), formula (L2-5) and formula (L2-6) can form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0590] n2 is an integer from 1 to 9;

[0591] m3 is an integer from 1 to 6;

[0592] n3 is an integer from 1 to 6;

[0593] j2 is an integer from 0 to 6);

[0594] Further preferably -L 2 - is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0595] [Chemical formula 77]

[0596]

[0597] (In formula (L2-3-1), formula (L2-4-1), formula (L2-5-1) and formula (L2-6-1), R A groups are each independently selected from a hydrogen atom, a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxyl C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca) or C 7～16 a group in aralkyl;

[0598] The -NH- group in formula (L2-3-1), formula (L2-4-1), formula (L2-5-1) and formula (L2-6-1) can form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom;

[0599] n2 is an integer from 1 to 5;

[0600] m3 is an integer from 1 to 3;

[0601] n3 is an integer from 1 to 4;

[0602] j2 is an integer from 0 to 3);

[0603] Particularly preferably a linking group selected from the following:

[0604] [Chemical formula 78]

[0605]

[0606] (In formula (L2-4-1), formula (L2-6-1) and formula (L2-6-1-a), R A groups are each independently selected from a hydrogen atom, a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxyl C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca) or C 7～16 a group in aralkyl;

[0607] n2 is an integer from 1 to 5;

[0608] m3 is an integer from 1 to 3;

[0609] n3 is an integer from 1 to 4;

[0610] j2 is an integer from 0 to 3;

[0611] k is an integer from 1 to 4).

[0612] [27-2] In formula (AM-2) of the foregoing method

[27] , P 1 is preferably an acetyl group or a benzoyl group, more preferably a benzoyl group.

[0613]

[28] The 28th method is as described below. An amino compound represented by the following formula (AM-2) or a pharmaceutically acceptable salt thereof:

[0614] [Chemical formula 79]

[0615]

[0616] [In formula (AM-2), -L 2 - is a linking group selected from the following partial structural formulas (in each formula, the outside of the dotted lines at both ends is not included):

[0617] [Chemical formula 80]

[0618]

[0619] (In formula (L2-a), R B is a group selected from a hydrogen atom, a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxyl C 1～6 alkyl or C 7～16 aralkyl);

[0620] P 1 is a benzoyl group or an acetyl group (wherein, when -L 2 - is formula (L2-b), P 1 is a benzoyl group).

[0621] [28-1] In formula (AM-2) of the foregoing method

[28] , -L 2 - is preferably a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0622] [Chemical formula 81]

[0623]

[0624] (In formula (L2-a), R B is a hydrogen atom, C 1～6 alkyl or C 7～16 aralkyl);

[0625] More preferably, it is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0626] [Chemical formula 82]

[0627]

[0628] (In formula (L2-a-1), R B is a hydrogen atom, C 1～6 alkyl or C 7～16 aralkyl);

[0629] More preferably, it is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0630] [Chemical formula 83]

[0631]

[0632] (In formula (L2-a-1), R B is a hydrogen atom, C 1～6 alkyl or C 7～16 aralkyl);

[0633] Particularly preferably, it is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0634] [Chemical formula 84]

[0635]

[0636] (In formula (L2-a-2), R B is a hydrogen atom, C 1～6 alkyl or C 7～16 aralkyl);

[0637] Particularly preferably, it is a linking group selected from the following partial structural formulas [in each formula, the outside of the dotted lines at both ends is not included]:

[0638] [Chemical formula 85]

[0639]

[0640] [28-2] In formula (AM-2) of the foregoing mode

[28] , P 1 is preferably benzoyl.

[0641]

[29] The 29th mode is as follows. An amino compound selected from the following formulas or a pharmaceutically acceptable salt thereof:

[0642] [Chemical formula 86]

[0643]

[0644] (In each formula, Bz is benzoyl).

[0645] [29-1] Among the amino compounds or pharmaceutically acceptable salts thereof of the aforementioned method

[29] , the amino compound is preferably an amino compound selected from the following formulas:

[0646] [Chemical formula 87]

[0647]

[0648] (In each formula, Bz is benzoyl).

[0649] 1. Alginate

[0650] In this specification, when it is described as alginate, it means at least one alginate selected from alginate, alginate ester, and their salts (for example, sodium alginate) (sometimes referred to as "alginate-like substances"). The alginate used can be derived from nature or can be a synthetic product, and preferably it is derived from nature. The preferably used alginate-like substances are biologically absorbable polysaccharides extracted from brown algae such as Macrocystis, Macrocystis pyrifera, Laminaria japonica, Ascophyllum nodosum, Cladosiphon okamuranus, Scytosiphon lomentaria, Ecklonia kurome, and Undaria pinnatifida, and are polymers in which two uronic acids, D-mannuronic acid (M) and L-glucuronic acid (G), are polymerized into a linear chain. More specifically, it is a block copolymer obtained by arbitrarily bonding the homopolymer component of D-mannuronic acid (MM component), the homopolymer component of L-glucuronic acid (GG component), and the component (M / G component) obtained by randomly arranging D-mannuronic acid and L-glucuronic acid.

[0651] In this specification, in the molecular weights of alginate, alginate derivatives, crosslinked alginate, and crosslinked alginate, as a unit, it is sometimes denoted as Da (Dalton).

[0652] In this specification, for alginate, alginate is denoted as (ALG), and any one carboxyl group of alginate is denoted as -COOH, and sometimes alginate is denoted as (ALG)-COOH.

[0653] In some embodiments, the alginate is sodium alginate. Commercially available sodium alginate can be used. For example, alginate having the physical property values shown in Table 1 can be used. Here, in Examples 10 to 12 described later, the sodium alginate used is the sodium alginate of A-2 shown in Table 1 (manufacturer: Morita Pharmaceutical Co., Ltd.). The viscosities, weight-average molecular weights, and M / G ratios of 1 w / w% aqueous solutions of each sodium alginate are shown in Table 1. In addition, in Examples 1 to 9 described later, the sodium alginate used is ALG-2 (seller: Kimika Co., Ltd.) and has the weight-average molecular weight described in <Measurement of molecular weight> described later.

[0654] [Table 1]

[0655]

[0656] The physical property values of the aforementioned sodium alginates A-1, A-2, A-3, B-1, B-2, and B-3 were measured using the following various methods. The measurement method is not limited to this method, and the physical property values may be different from the above depending on the measurement method.

[0657] [Measurement of Viscosity of Sodium Alginate]

[0658] According to the viscosity measurement method in the Japanese Pharmacopoeia (16th Edition), the measurement was carried out using the rotational viscometer method (cone-plate type rotational viscometer). The specific measurement conditions are as follows. The sample solution was prepared using MilliQ water. The measuring instrument used was a cone-plate type rotational viscometer (viscosity viscoelasticity measuring device Rheostress RS600 (Thermo Haake GmbH), sensor: 35 / 1). The rotation speed was 1 rpm when measuring the 1 w / w% sodium alginate solution. The reading time was set to 2 minutes of measurement, and the average value from 1 minute to 2 minutes from the start was taken. The average value of three measurements was used as the measured value. The measurement temperature was set to 20 °C.

[0659] [Measurement of Weight-Average Molecular Weight of Sodium Alginate]

[0660] It was measured using two measurement methods: (1) gel permeation chromatography (GPC) and (2) GPC-MALS. The measurement conditions are as follows.

[0661] [Pretreatment Method]

[0662] After adding the eluent to the sample and dissolving it, the solution obtained by filtering through a 0.45 μm membrane filter was used as the measurement solution.

[0663] (1) Gel Permeation Chromatography (GPC) Measurement

[0664] [Measurement Conditions (Relative Molecular Weight Distribution Measurement)]

[0665] Column: TSKgel GMPW-XL × 2 + G2500PW-XL (7.8 mm I.D. × 300 mm × 3 columns)

[0666] Eluent: 200 mM sodium nitrate aqueous solution

[0667] Flow rate: 1.0 mL / min

[0668] Concentration: 0.05%

[0669] Detector: RI detector

[0670] Column temperature: 40 °C

[0671] Injection volume: 200 μL

[0672] Molecular weight standards: Standard amylopectin, glucose.

[0673] (2) GPC-MALS measurement

[0674] [Measurement of refractive index increment (dn / dc) (measurement conditions)]

[0675] Differential refractive index meter: Optilab T-rEX

[0676] Measurement wavelength: 658 nm

[0677] Measurement temperature: 40 °C

[0678] Solvent: 200 mM aqueous sodium nitrate solution

[0679] Sample concentration: 0.5 - 2.5 mg / mL (5 concentrations).

[0680] [Measurement conditions (absolute molecular weight distribution measurement)]

[0681] Column: TSKgel GMPW-XL × 2 + G2500PW-XL (7.8 mm I.D. × 300 mm × 3 columns)

[0682] Eluent: 200 mM aqueous sodium nitrate solution

[0683] Flow rate: 1.0 mL / min

[0684] Concentration: 0.05%

[0685] Detector: RI detector, light scattering detector (MALS)

[0686] Column temperature: 40 °C

[0687] Injection volume: 200 μL.

[0688] In this specification, in the molecular weights of alginic acid, alginic acid derivatives, crosslinked alginic acid, and crosslinked alginic acid, the unit is sometimes denoted as Da (Dalton).

[0689] The composition ratio (M / G ratio) of D-mannuronic acid and L-guluronic acid in alginic acids varies mainly depending on the species of organisms such as seaweeds. In addition, it is affected by the growth location and season of the organisms, covering a wide range from a high G type with an M / G ratio of about 0.2 to a high M type with an M / G ratio of about 5. The gelation ability of alginic acids and the properties of the gels formed are affected by the M / G ratio. Generally, it is known that the gel strength increases when the G ratio is high. In addition, the M / G ratio also affects the hardness, brittleness, water absorbency, softness, etc. of the gels. The M / G ratio of the alginic acids and / or their salts used is usually 0.2 - 4.0, more preferably 0.4 - 3.0, and further preferably 0.5 - 3.0.

[0690] In this specification, the numerical range expressed using "~" indicates a range that includes the numerical values described before and after "~" as the respective minimum and maximum values.

[0691] In this specification, the "alginic acid ester" and "alginic acid salt" used are not particularly limited. In order to react with a crosslinking agent, it is necessary not to have a functional group that hinders the crosslinking reaction. As the alginic acid ester, propylene glycol alginate etc. are preferably selected.

[0692] In this specification, examples of the alginic acid salt include a monovalent salt of alginic acid and a divalent salt of alginic acid. As the monovalent salt of alginic acid, sodium alginate, potassium alginate, ammonium alginate etc. are preferably selected, more preferably sodium alginate or potassium alginate, and particularly preferably sodium alginate. As the divalent salt of alginic acid, calcium alginate, magnesium alginate, barium alginate, strontium alginate etc. are preferably selected.

[0693] Alginic acid is a high molecular polysaccharide, and it is difficult to accurately determine the molecular weight. Generally, the range of the weight average molecular weight is 1,000 to 10 million, preferably 10,000 to 8 million, and more preferably 20,000 to 3 million. In the measurement of the molecular weight of a high molecular substance derived from a natural product, it is known that there are differences in values depending on the measurement method.

[0694] For example, the weight average molecular weight measured by gel permeation chromatography (GPC) or gel filtration chromatography (they are also collectively referred to as size exclusion chromatography) is preferably 100,000 or more, more preferably 500,000 or more, and preferably 5 million or less, more preferably 3 million or less. The preferred range is 100,000 to 5 million, and more preferably 150,000 to 3 million.

[0695] In addition, for example, according to the GPC-MALS method, the absolute weight average molecular weight can be measured. The weight average molecular weight (absolute molecular weight) measured by the GPC-MALS method is preferably 10,000 or more, more preferably 50,000 or more, further preferably 60,000 or more, and preferably 1 million or less, more preferably 800,000 or less, further preferably 700,000 or less, and particularly preferably 500,000 or less. The preferred range is 10,000 to 1 million, more preferably 50,000 to 800,000, further preferably 60,000 to 700,000, and particularly preferably 60,000 to 500,000.

[0696] Usually, when calculating the molecular weight of a high molecular polysaccharide using the above method, a measurement error of 10% to 20% occurs. For example, if it is 400,000, the value fluctuates in the range of about 320,000 to 480,000. If it is 500,000, the value fluctuates in the range of about 400,000 to 600,000. If it is 1 million, the value fluctuates in the range of about 800,000 to 1.2 million.

[0697] The molecular weight of alginic acid substances can be measured according to conventional methods.

[0698] Representative conditions when gel filtration chromatography is used in molecular weight determination are as described in the examples of this specification below. For the column, for example, a Superose6 Increase 10 / 300GL column (GE Healthcare) can be used. As the developing solvent, for example, a 10 mmol / L phosphate buffer (pH 7.4) containing 0.15 mol / L NaCl can be used. As the molecular weight standards, blue dextran, thyroglobulin, ferritin, aldolase, conalbumin, ovalbumin, ribonuclease A, and aprotinin can be used.

[0699] The viscosity of the alginic acid used in this specification is not particularly limited. When the viscosity is measured in the form of an aqueous solution of 1 w / w% alginic acid-based substances, it is preferably 10 mPa·s to 1000 mPa·s, more preferably 50 mPa·s to 800 mPa·s.

[0700] The viscosity of the aqueous solution of alginic acid can be measured according to conventional methods. For example, a coaxial double-cylinder type rotational viscometer, a single-cylinder type rotational viscometer (Brookfield type viscometer), a cone-plate type rotational viscometer (cone-plate viscometer), etc. of the rotational viscometer method can be used for measurement. It is preferably measured according to the viscosity measurement method of the Japanese Pharmacopoeia (16th edition). More preferably, a cone-plate viscometer is used.

[0701] Alginic acid-based substances are obtained by extracting from brown algae. Initially, they have a large molecular weight and high viscosity, but through processes such as drying and purification based on heat, the molecular weight becomes smaller and the viscosity becomes lower. Alginic acid-based substances with different molecular weights can be prepared by methods such as controlling conditions such as the temperature in the preparation step, selecting brown algae as the raw material, and fractionating the molecular weight in the preparation step. Further, by mixing with other types of alginic acid-based substances having different molecular weights or viscosities, alginic acid-based substances with the target molecular weight can also be formed.

[0702] The alginic acid used in this specification is, in some cases, alginic acid that has not been treated for low endotoxin, or in other cases, alginic acid that has been treated for low endotoxin. Low endotoxin means that the endotoxin level is low enough not to substantially cause inflammation or fever. More preferably, alginic acid-based substances that have been treated for low endotoxin are desired.

[0703] The low endotoxin treatment can be carried out by known methods or methods based thereon. For example, it can be carried out by the method of Suga et al. for purifying sodium hyaluronate (for example, refer to Japanese Patent Laid-Open No. 9-324001, etc.), the method of Yoshida et al. for purifying β1,3-glucan (for example, refer to Japanese Patent Laid-Open No. 8-269102, etc.), the method of William et al. for purifying biopolymer salts such as sodium alginate and gellan gum (for example, refer to Japanese Patent Application Laid-Open No. 2002-530440, etc.), the method of James et al. for purifying polysaccharides (for example, refer to International Publication No. 93 / 13136 pamphlet, etc.), the method of Louis et al. (for example, refer to the specification of U.S. Patent No. 5589591, etc.), the method of Hermann Frank et al. for purifying alginate (for example, refer to Appl Microbiol Biotechnol (1994) 40: 638-643, etc.) or methods based on them. The low endotoxin treatment is not limited to them, and can be carried out by known methods such as washing, filtration based on filters (endotoxin-removing filters, charged filters, etc.), ultrafiltration, purification using columns (endotoxin adsorption affinity columns, gel filtration columns, columns based on ion exchange resins, etc.), adsorption in hydrophobic substances, resins or activated carbon, organic solvent treatment (extraction based on organic solvents, precipitation / sedimentation caused by adding organic solvents, etc.), surfactant treatment (for example, refer to Japanese Patent Laid-Open No. 2005-036036, etc.), or by appropriately combining them. Known methods such as centrifugation can be appropriately combined in these treatment steps. It is desirable to appropriately select according to the type of alginic acid.

[0704] The endotoxin level can be confirmed by known methods. For example, it can be measured by the method using Limulus reagent (LAL), the method using Endospecy (registered trademark) ES-24S Set (Seikagaku Corporation), etc.

[0705] There is no particular limitation on the treatment method of the endotoxin used. As a result, when performing endotoxin measurement using Limulus reagent (LAL), the endotoxin content of alginic acid is preferably 500 endotoxin units (EU) / g or less, more preferably 100 EU / g or less, particularly preferably 50 EU / g or less, and especially preferably 30 EU / g or less. The low endotoxin-treated sodium alginate can be obtained through commercially available products such as Sea Matrix (registered trademark) (Morita Pharmaceutical Co., Ltd.), PRONOVA TM UP LVG (FMC BioPolymer), etc.

[0706] 2. Alginic acid derivatives

[0707] In this specification, an alginic acid derivative can be provided. In this specification, as the alginic acid derivative, a reactive group in a Michael addition reaction or a reactive group complementary to the reactive group is introduced through an amide bond and a divalent linking group on any one or more carboxyl groups of alginic acid.

[0708] More specifically, they are the alginic acid derivative represented by the following formula (I) and the alginic acid derivative represented by the following formula (II):

[0709] [Chemical Formula 88]

[0710]

[0711] [In formula (I), the definitions of (ALG), -CONH-, and -L 1 - are the same as those defined in the foregoing method [1] or [2]],

[0712] [Chemical Formula 89]

[0713]

[0714] [In formula (II), the definitions of (ALG), -CONH-, -L 2 - are the same as those defined in the foregoing method [1] or [5]].

[0715] The reactive group of the alginic acid derivative represented by formula (I) has an acrylic acid residue, and the reactive group of the alginic acid derivative represented by formula (II) (the reactive group complementary to the reactive group of the alginic acid derivative represented by formula (I)) is a thiol residue. These two reactive groups, the acrylic acid residue and the thiol residue, can easily form a covalent bond through a Michael addition reaction.

[0716] Examples of the acrylic acid residue include residues that can form an adduct through a Michael addition reaction with a thiol residue. Specifically, examples include an acryloyl group, an acrylic acid group, a maleoyl group, a maleimide group, a fumaric group, etc. Preferably, it is an acryloyl group or a maleimide group, and more preferably a maleimide group.

[0717] Examples of the thiol residue include residues that can form an adduct through a Michael addition reaction with an acrylic acid residue. Specifically, examples include a benzylthiol group, a benzenethiol group, an alkylthiol group (for example, a thiol group such as a methanethiol residue, an ethanethiol residue, a cysteine residue, etc. substituted with a C 1～6 alkyl residue), etc. Preferably, it is a benzylthiol group or an alkylthiol group, and more preferably an alkylthiol group.

[0718] The aforementioned divalent linking group (-L 1 - or -L 2-) Any linear group can be used in such a way that the reactive group or the complementary reactive group is kept at a certain distance from alginic acid as long as it does not hinder the reaction between the reactive group and the complementary reactive group. As the divalent linking group, for example, a linear alkylene group (-(CH 2 ) n -) (where n = 1 to 30) can be mentioned. In the alkylene group, -CH 2 - can be substituted by a plurality of (for example, 1 to 10 or 1 to 5) -C(=O)-, -CONH-, -O-, -NH-, -S-, benzene ring, heterocyclic ring (5- to 6-membered aromatic heterocyclic ring or 5- to 6-membered non-aromatic heterocyclic ring such as pyridine ring, piperidine ring, piperazine ring, etc.); the hydrogen atom of -CH 2 - can be selected from an oxo group (=O), a halogen atom, a hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group (M is Li, Na, K or 1 / 2Ca), -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl (in the aforementioned -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, R a and R b are each independently selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl), guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl) and are substituted by a plurality of groups (for example, 1 to 10 or 1 to 5); when two hydrogen atoms of the same -CH 2 - are substituted by C 1～6 alkyl, the alkyls can be bonded to each other to form C 3～8cycloalkyl ring), but not limited to them.

[0719] Specifically, regarding -L 1 -, it is a divalent linking group described in the aforementioned method [1] or [2]. Regarding -L 2 -, a divalent linking group described in the aforementioned method [1] or [5] can be cited.

[0720] In this specification, for the linking group -L 1 - (in formulae (L1-1) to (L1-4)), when there is a chiral carbon in the formula, it means that each of its optical isomers is also included.

[0721] For example, when -L 1 - in formula (I) is the following formula (L1-3-1-a) (where the two outer sides of the dotted line are not included):

[0722] [Chemical formula 90]

[0723]

[0724] It means a linking group shown by the following formula (L1-3-1-aS) in which the stereoconfiguration of the carbon substituted by benzyl is the S-form and the following formula (L1-3-1-aR) in which the stereoconfiguration of the carbon substituted by benzyl is the R-form (in any formula, the two outer sides of the dotted line are not included):

[0725] [Chemical formula 91]

[0726]

[0727] In addition, similarly, for the linking group -L 2 - (in formulae (L2-1) to (L2-6)) in formula (II) or formula (II-P), when there is a chiral carbon in the formula, it means that each of its optical isomers is also included.

[0728] For example, when -L 2 - in formula (II) or formula (II-P) is the following formula (L2-6-1-a) (where the two outer sides of the dotted line are not included):

[0729] [Chemical formula 92]

[0730]

[0731] It means a linking group shown by the following formula (L2-6-1-aS) in which the stereoconfiguration of the carbon substituted by benzyl is the S-form and the following formula (L2-6-1-aR) in which the stereoconfiguration of the carbon substituted by benzyl is the R-form (in any formula, the two outer sides of the dotted line are not included):

[0732] [Chemical formula 93]

[0733]

[0734] In formula (I) in the present specification, when there is a chiral carbon in the linking group -L 1 - (when it is an optically active substance), in the step of synthesizing the amine derivative (AM-1) corresponding to formula (I), each optically active substance can be separated from its racemate by a usual optical resolution means (separation method), and in the step of synthesizing the amine derivative (AM-1) corresponding to formula (I), one of the optical isomers can be selectively synthesized by using asymmetric synthesis, and thus each optically active substance can be synthesized. By using the obtained optically active amine derivatives, an alginic acid derivative of formula (I) having a chiral carbon (optically active) can be synthesized. Further, in formula (II) or formula (II-P) in the present specification, when there is a chiral carbon in the linking group -L 2 -, in the step of synthesizing the amine derivative (AM-2) corresponding to formula (II) or formula (II-P) respectively, in the same manner as the above-described method, each optically active substance and an alginic acid derivative of formula (II) or formula (II-P) having a chiral carbon (optically active) can be synthesized.

[0735] As the above-described separation method, for example, optical resolution methods such as fractional recrystallization method, diastereomer method, and chiral column method can be mentioned. Each resolution method will be described in detail below.

[0736] Fractional recrystallization method: A method in which a racemate is ionically bonded to an optical resolving agent to obtain crystalline diastereomers, and then the crystalline diastereomers are separated by fractional recrystallization method, and an optically pure compound is obtained through a step of removing the optical resolving agent as needed. Examples of the optical resolving agent include (+)-mandelic acid, (-)-mandelic acid, (+)-tartaric acid, (-)-tartaric acid, (+)-1-phenylethylamine, (-)-1-phenylethylamine, cinchonine, (-)-cinchonidine, and strychnine.

[0737] Diastereomer method: A reaction in which a mixture of racemates is covalently bonded to an optical resolving agent to obtain a mixture of diastereomers, and then the mixture is separated into optically pure diastereomers by usual separation means (for example, fractional recrystallization, silica gel column chromatography, and HPLC, etc.), and then an optically pure optical isomer is obtained through a step of removing the optical resolving agent based on a chemical reaction (hydrolysis reaction, etc.).

[0738] For example, when the compound or intermediate compound in this specification has a hydroxyl group or an amino group (primary or secondary), diastereomers of an ester form or an amide form can be obtained respectively through the condensation reaction of the compound with an optically active organic acid (for example, α-methoxy-α-(trifluoromethyl)phenylacetic acid, (-)-menthyloxyacetic acid, etc.). And when the compound in this specification has a carboxyl group, diastereomers of an amide form or an ester form can be obtained respectively through the condensation reaction of the compound with an optically active amine or an optically active alcohol. The diastereomers obtained through the condensation reaction are separated, and each diastereomer is subjected to an acid- or base-based hydrolysis reaction, thereby converting into an optically pure optical isomer of the original compound.

[0739] Chiral column method: A method of directly performing optical resolution by subjecting a racemate or its salt to chromatography based on a chiral column (column for separating optical isomers).

[0740] For example, in the case of high performance liquid chromatography (HPLC), a mixture of optical isomers is added to a chiral column (for example, CHIRAL series manufactured by Daicel Corporation, etc.), and developed using an elution solvent (water, various buffers (for example, phosphate buffer), and organic solvents (for example, ethanol, methanol, isopropanol, acetonitrile, trifluoroacetic acid, and diethylamine, etc.), etc., either individual solvents or their mixed solvents), whereby the optical isomers can be separated. And, for example, in the case of gas chromatography, optical isomers can be separated using a chiral column (for example, CP-Chirasil-DeX CB (manufactured by GL Sciences Inc., etc.)). And, for example, in the case of supercritical fluid chromatography (SFC), a mixture of optical isomers is added to a chiral column (for example, CHIRAL series manufactured by Daicel Corporation, etc.), and carbon dioxide and an appropriate organic solvent (for example, methanol, ethanol, isopropanol, trifluoroacetic acid, and diethylamine, etc.) are used as the elution solvent, whereby the optical isomers can be separated.

[0741] Asymmetric synthesis for selectively synthesizing one of the aforementioned optical isomers includes (1) an asymmetric synthesis reaction in which a racemic compound reacts enantioselectively to introduce an optically active substance, (2) a method of selectively synthesizing diastereomers from naturally occurring optically active compounds (sugars, amino acids, etc.), etc.

[0742] The alginic acid derivatives represented by formula (I) and formula (II) as alginic acid derivatives in this specification can be prepared, for example, by the method of the following formula (for details, refer to the general preparation method described later).

[0743] [Chemical formula 94]

[0744]

[0745] The weight-average molecular weight of the alginic acid derivative represented by formula (I), formula (II) or formula (II-P) in this specification is 100,000 Da to 3,000,000 Da, preferably 300,000 Da to 2,500,000 Da, and more preferably 500,000 Da to 2,000,000 Da. The molecular weight of the two alginic acid derivatives can be determined by the method described below.

[0746] In this specification, for the following formula (BR-1) in the alginic acid derivative represented by formula (I):

[0747] [Chemical formula 95]

[0748] The group does not need to be bonded to all the carboxyl groups of the alginic acid constituent unit. And, for the following formula (BR-2) in the alginic acid derivative represented by formula (II):

[0749] [Chemical formula 96]

[0750] The group does not need to be bonded to all the carboxyl groups of the alginic acid constituent unit. And, for the following formula (BR-2-P) in the alginic acid derivative represented by formula (II-P):

[0751] [Chemical formula 97]

[0752] The group does not need to be bonded to all the carboxyl groups of the alginic acid constituent unit.

[0753] In this specification, when the group of formula (BR-1) in the alginic acid derivative of formula (I) is referred to as a reactive group, the group of formula (BR-2) in the alginic acid derivative of formula (II) becomes a complementary reactive group. And, conversely, when the group of formula (BR-2) in the alginic acid derivative of formula (II) is referred to as a reactive group, the group of formula (BR-1) in the alginic acid derivative of formula (I) becomes a complementary reactive group.

[0754] In this specification, the introduction rate of the reactive group or the complementary reactive group is, for example, 0.1% to 30% or 1% to 30% respectively, preferably 2% to 20%, and more preferably 3% to 10%.

[0755] The introduction rate of the aforementioned reactive group or complementary reactive group is a value expressed as a percentage of the number of uronic acid monosaccharide units in which each reactive group is introduced among the uronic acid monosaccharide units that are the repeating units of alginic acid. In this specification, unless otherwise specified, the % used for the introduction rate of the reactive group or complementary reactive group in the alginic acid derivative (formula (I) or formula (II)) refers to mol%. The introduction rate of each reactive group or complementary reactive group can be determined by the method described in the examples below.

[0756] The protecting group P for the thiol group in the alginic acid derivative represented by formula (II-P) 1 A protecting group that can be easily protected and deprotected can be appropriately selected. For example, a protecting group known from the literature described in "Protective Groups in Organic Synthesis 5th Edition, 2014, John Wiley & Sons, Greene, etc." can be appropriately selected. More specifically, as the protecting group P 1 , examples include C 2～6 alkanoyl groups such as acetyl group and ethylcarbonyl group; C 6～10 arylcarbonyl groups such as benzoyl group and naphthylcarbonyl group; trityl (triphenylmethyl), diphenylmethyl; N-C 1～6 alkyl-carbamoyl groups such as methylaminocarbonyl group and ethylaminocarbonyl group, but are not limited to them.

[0757] In the present specification, the maleimide group in the group of formula (BR-1) in the alginic acid derivative of formula (I) and the thiol (HS-) group in the alginic acid derivative of formula (II) can form a covalent bond through a Michael addition reaction, thereby forming a crosslink.

[0758] 3. Method for synthesizing alginic acid derivative

[0759] In the present specification, the alginic acid derivative represented by formula (I) can be prepared by a condensation reaction of an amine derivative represented by formula (AM-1) (where -L 1 - is the same as defined in the foregoing method [1] or [2]) or a salt thereof with any carboxyl group of an alginic acid. And, the alginic acid derivative represented by formula (II) can be obtained by a condensation reaction of an amine derivative represented by formula (AM-2) (where -L 2 - and P 1 are the same as defined in the foregoing method [1] or [5]) or a salt thereof with any carboxyl group of an alginic acid to obtain the alginic acid derivative represented by formula (II-P), and then by deprotection of the protecting group P 1 group.

[0760] [Chemical formula 98]

[0761]

[0762] Specifically, an aqueous solution of alginic acid at 0.5% to 1% by weight and an amine derivative represented by the formula (AM-1) or (AM-2) or a salt thereof are used. According to the methods known from the literature, for example, the methods described in "Experimental Chemistry Course, 5th Edition, 16, Synthesis of Organic Compounds IV, Carboxylic Acids and Derivatives, Acid Amides and Acid Imides, pages 118 - 154, Amino Acids · Peptides, pages 258 - 283, 2007, Maruzen", etc., in the presence of a condensing agent such as 1,3 - dicyclohexylcarbodiimide (DCC), 1 - ethyl - 3 - (3 - dimethylaminopropyl)carbodiimide hydrochloride (WSC·HCl), benzotriazol - 1 - yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP reagent), bis(2 - oxo - 3 - oxazolidinyl)phosphinic chloride (BOP - Cl), 2 - chloro - 1,3 - dimethylimidazolinium hexafluorophosphate (CIP), 4 - (4,6 - dimethoxy - 1,3,5 - triazin - 2 - yl) - 4 - methylmorpholinium chloride (DMT - MM), etc., in water, an ether - based solvent such as 1,4 - dioxane, an alcohol - based solvent such as methanol or ethanol, a polar solvent such as N,N - dimethylformamide or a mixed solvent thereof (wherein the mixed solvent is a mixed solvent in which alginic acid does not precipitate), in the presence or absence of an inorganic base such as sodium bicarbonate or sodium carbonate or an organic base such as triethylamine or pyridine, the reaction is carried out at a temperature of 0°C to 50°C, whereby an alginic acid derivative represented by the formula (I) or (II - P) can be prepared. Further, by deprotecting the protecting group P 1 of the formula (II - P), an alginic acid derivative represented by the formula (II) can be prepared.

[0763] When the P 1 group of the formula (II - P) is an acyl group such as an acetyl group or a benzoyl group or a protecting group of the benzoyl type, the deprotection can be carried out by the following method. To an aqueous solution (for example, 0.5% to 1% by weight) of the alginic acid derivative of the formula (II - P), the following formula (BR - 2 - P) is introduced, 2～6 in an excessive amount, an inorganic base such as sodium hydroxide or potassium hydroxide is added, and hydrolysis is carried out at a reaction temperature of 0°C to 30°C, whereby a salt of the alginic acid derivative of the formula (II) can be prepared. Then, by neutralizing the reaction solution, it can be directly used as a solution of the alginic acid derivative of the formula (II) for the cross - linking reaction.

[0764] [Chemical Formula 99]

[0765]

[0766]

[0767] ​In the method for preparing the alginic acid derivative of the aforementioned formula (I) or the alginic acid derivative of formula (II-P), the introduction rate of the amine derivative of formula (AM-I) or formula (AM-II) can be adjusted by appropriately selecting reaction conditions such as the following (i) to (v) and combining them in consideration of the properties of the amine, etc. For (i) increasing or decreasing the equivalent amount of the condensing agent, (ii) raising or lowering the reaction temperature, (iii) extending or shortening the reaction time, (iv) adjusting the concentration of the alginic acid as the reaction substrate, (v) increasing the solubility of the amino derivative of formula (AM-1) or formula (AM-2), an organic solvent mixed with water is added, etc.

[0768] The following shows the method for preparing the amine derivative represented by formula (AM-1) or formula (AM-2).

[0769] [Preparation method A] <Synthesis method of the amine derivative of formula (AM-1) and its salt>

[0770] [Chemical formula 100]

[0771]

[0772] [Preparation method A] <Step 1>

[0773] Using the amine represented by formula (III) [the compound of formula (III) is a commercially available compound or a compound that can be prepared from commercially available compounds by a preparation method known from the literature. In the formula, P 2 is a protecting group for the amino group and can be appropriately selected] and maleic acid, according to a method known from the literature, such as the method described in "Experimental Chemistry Course 5th Edition 16, Synthesis of Organic Compounds IV, Carboxylic Acids and Derivatives, Acid Amides and Acid Imides, pages 146 - 154, 2007, Maruzen", etc., in a solvent selected from ether solvents such as tetrahydrofuran and 1,4-dioxane, halogen solvents such as dichloromethane and 1,2-dichloroethane, polar solvents such as N,N-dimethylformamide, etc., in the presence of a condensing agent selected from 1,3-dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (WSC·HCl), benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP reagent), bis(2-oxo-3-oxazolidinyl)phosphinic chloride (BOP-Cl), 2-chloro-1,3-dimethylimidazolinium hexafluorophosphate (CIP), 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM), etc., in the presence or absence of a base selected from inorganic bases such as sodium bicarbonate, sodium carbonate, potassium carbonate, etc. or organic bases such as triethylamine, N,N-diisopropylethylamine, pyridine, etc., and reacting at a temperature between 0 °C and 50 °C, whereby the compound of formula (IV) can be prepared.

[0774] Furthermore, the compound of formula (IV) can be prepared by reacting maleic anhydride with the amine represented by formula (III) in a solvent such as an alcohol-based solvent like methanol or ethanol, in the presence or absence of a base such as triethylamine, N,N-diisopropylethylamine, or pyridine.

[0775] [Preparation Method A] <Step 2>

[0776] The compound of formula (VI) can be prepared by reacting the compound represented by formula (IV) with a base such as sodium acetate in a solvent selected from ether-based solvents such as tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, hydrocarbon-based solvents such as toluene, benzene, xylene, halogen-based solvents such as 1,2-dichloroethane, dichloromethane, chloroform, or acetic anhydride, at a temperature between 40 °C and the reflux temperature of the solvent.

[0777] Also, the compound of formula (VI) can be prepared by reacting the crude compound of formula (IV) obtained in [Preparation Method A] <Step 1> with a base such as sodium acetate in acetic anhydride.

[0778] Also, the compound of formula (VI) can be prepared by reacting the compound or crude compound of formula (IV) obtained in [Preparation Method A] <Step 1> with an appropriate condensing agent to derivatize it into an active ester form and then performing a cyclization reaction.

[0779] [Preparation Method A] <Step 3>

[0780] The compound of formula (VI) can be prepared by reacting the compound represented by formula (V) [the compound of formula (V) is a commercially available compound or a compound that can be prepared from commercially available compounds using preparation methods known from the literature.] with maleimide in a solvent selected from ether-based solvents such as tetrahydrofuran, 1,4-dioxane, hydrocarbon-based solvents such as toluene, benzene, xylene, etc., in the presence of a phosphine reagent such as triphenylphosphine and Mitsunobu reagents such as diethyl azodicarboxylate and diisopropyl azodicarboxylate, at a temperature between -78 °C and the reflux temperature of the solvent.

[0781] [Preparation Method A] <Step 4>

[0782] The compound represented by formula (VI) is deprotected according to the method for deprotecting the protecting group of the amino group described in the literature, such as "Protective Groups in Organic Synthesis 5th Edition, 2014, John Wiley & Sons, Greene, etc.", according to the type of the protecting group P 2 to obtain the amine derivative of formula (AM-1).

[0783] The amine derivative of formula (AM-1) can be obtained in the form of a salt as needed. For example, salts such as hydrochloride and trifluoroacetate can be cited.

[0784] P in [Preparation Method A] 2 represents a protecting group for the amine. For example, the protecting groups described in "Protective Groups in Organic Synthesis 5th Edition, 2014, John Wiley & Sons, Greene, etc." can be appropriately selected. As P 2 , for example, -C(O)O-tertBu group, -C(O)O-Bn group, -C(O)O-CH 2 -CH=CH 2 group, -C(O)CH 3 group, -C(O)CF 3 group, -SO 2 Ph, -SO 2 PhMe group, -SO 2 Ph(NO 2 ) group and other protecting groups, but not limited to them.

[0785] P 2 For example, when it is -C(O)O-tertBu group, deprotection can be carried out by using an acid (hydrogen chloride (it can be a solution of hydrogen chloride in 1,4-dioxane, cyclopentyl methyl ether, ethyl acetate, etc.), trifluoroacetic acid, etc.). More specifically, for example, refer to the methods known from documents such as "Protective Groups in Organic Synthesis 5th Edition, 2014, John Wiley & Sons, Greene, etc.", and select the deprotection method according to the type of protecting group.

[0786] [Preparation Method B] Synthesis method of the amine derivative of formula (AM-2) and its salt (Reaction Formula B)

[0787] [Chemical Formula 101]

[0788]

[0789] [Preparation Method B] <Step 1>

[0790] Use formula (VIII) [The compound of formula (VIII) is a commercially available compound or a compound that can be prepared from commercially available compounds by the preparation methods known from documents. In the formula, P 3P, which is a protecting group for the amino group, can be appropriately selected. The compound shown by [] is used, and according to the methods known from the literature, such as the methods described in "Protective Groups in Organic Synthesis, 3rd Edition, PROTECTION FOR THE THETHIOL GROUP, pages 457 - 486, 1999", etc., the protecting group P is introduced. 1 , whereby the compound of formula (IX) can be prepared. For example, 1 when P is an acetyl group, acetyl chloride can be used; when P is a benzoyl group, benzoyl chloride can be used; when P is a trityl group, trityl chloride can be used; when P is an EtNHCO - group, ethyl isocyanate can be used, and thus the protecting group can be introduced.

[0791] Moreover, by using the compound shown by formula (VIII) and a carboxylic acid derivative such as acetic acid or benzoic acid, a condensation reaction is carried out according to the method of [Preparation Method A] <Step 1>, whereby the compound of formula (IX) can be prepared.

[0792] [Preparation Method B] <Step 2>

[0793] Using the compound shown by formula (XI) [the compound of formula (XI) is a commercially available compound or a compound that can be prepared from commercially available compounds by methods known from the literature. In the formula, P 3 is a protecting group for the amino group and can be appropriately selected. X can be selected from halogen atoms, such as a chlorine atom, a bromine atom, and an iodine atom.] and an acylthio derivative such as thiobenzoic acid, thioacetic acid, or potassium thioacetate, etc., in a solvent selected from acetonitrile, dichloromethane, N,N - dimethylformamide, etc., in the presence or absence of a base such as potassium carbonate, a reaction is carried out, whereby the compound of formula (IX) can be prepared.

[0794] [Preparation Method B] <Step 3>

[0795] The compound shown by formula (IX) is deprotected according to the methods for deprotecting the protecting group of the amino group known from the literature, such as those described in "Protective Groups in Organic Synthesis 5th Edition", 5th Edition, 2014, John Wiley & Sons, Greene, etc., according to the type of the protecting group P 3 , whereby the amine derivative of formula (AM - 2) can be prepared.

[0796] The amine derivative of formula (AM - 2) can be obtained in the form of a salt as needed. For example, salts such as hydrochloride and trifluoroacetate can be mentioned.

[0797] P in [Preparation Method B] 3Represents a protecting group for an amine. For example, a protecting group described in "Protective Groups in Organic Synthesis 5th Edition, 2014, John Wiley & Sons, Greene et al." can be appropriately selected. As P 3 , for example, is a -C(O)O-tertBu group, a -C(O)O-Bn group, a -C(O)O-CH 2 -CH=CH 2 group, a -C(O)CH 3 group, a -C(O)CF 3 group, a -SO 2 Ph, a -SO 2 PhMe group, a -SO 2 Ph(NO 2 ) group and other protecting groups, but is not limited to them.

[0798] P 3 When it is, for example, a -C(O)O-tertBu group, deprotection can be carried out by using an acid (hydrogen chloride (which can be a solution of hydrogen chloride in 1,4-dioxane, cyclopentyl methyl ether, ethyl acetate, etc.), trifluoroacetic acid, etc.). More specifically, for example, a method known from documents such as "Protective Groups in Organic Synthesis 5th Edition, 2014, John Wiley & Sons, Greene et al." can be referred to, and the deprotection method can be selected according to the type of the protecting group.

[0799] In this specification, when the amine derivative (amino compound) represented by formula (AM-1) or formula (AM-2) (including the sub-formulas of each formula) has a carboxyl group substituted thereon, it may sometimes form a pharmaceutically acceptable salt (for example, a base addition salt). Such salts are not particularly limited as long as they are pharmaceutically acceptable salts, and examples thereof include metal salts, ammonium salts, salts with organic bases, and the like. Suitable examples of metal salts include, for example, alkali metal salts such as lithium salt, sodium salt, potassium salt, cesium salt, alkaline earth metal salts such as calcium salt, magnesium salt, barium salt, and aluminum salt. Suitable examples of salts with organic bases include, for example, salts with methylamine, ethylamine, tert-butylamine, tert-octylamine, diethylamine, trimethylamine, triethylamine, cyclohexylamine, dicyclohexylamine, diphenylmethylamine, ethanolamine, diethanolamine, triethanolamine, piperidine, morpholine, pyridine, methylpyridine, lysine, arginine, ornithine, ethylenediamine, N-methylglucosamine, glucosamine, phenylglycine alkyl ester, guanidine, 2,6-dimethylpyridine, ethanolamine, diethanolamine, triethanolamine, N,N'-diphenylmethylethylenediamine, and the like. Among them, pharmaceutically acceptable salts are preferred.

[0800] In this specification, when the amine derivative (amino compound) represented by formula (AM-1) or formula (AM-2) (including the sub-formulas of each formula) has a carboxyl group substituted thereon, it may sometimes form a pharmaceutically acceptable salt (for example, a base addition salt). Such salts are not particularly limited as long as they are pharmaceutically acceptable salts, and examples thereof include metal salts, ammonium salts, salts with organic bases, and the like. Suitable examples of metal salts include, for example, alkali metal salts such as lithium salt, sodium salt, potassium salt, cesium salt, alkaline earth metal salts such as calcium salt, magnesium salt, barium salt, and aluminum salt. Suitable examples of salts with organic bases include, for example, salts with methylamine, ethylamine, tert-butylamine, tert-octylamine, diethylamine, trimethylamine, triethylamine, cyclohexylamine, dicyclohexylamine, diphenylmethylamine, ethanolamine, diethanolamine, triethanolamine, piperidine, morpholine, pyridine, methylpyridine, lysine, arginine, ornithine, ethylenediamine, N-methylglucosamine, glucosamine, phenylglycine alkyl ester, guanidine, 2,6-dimethylpyridine, ethanolamine, diethanolamine, triethanolamine, N,N'-diphenylmethylethylenediamine, and the like.

[0801] According to conventional methods, for example, by mixing the compounds of the present invention and a solution containing an appropriate amount of acid or base, forming the target salt and then filtering or distilling off the mixed solvent respectively, the aforementioned salt can be obtained. As a general discussion on salts, Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Stahl & Wermuth (Wiley-VCH, 2002) has been published, and it is described in detail in this book.

[0802] In this specification, the amine compounds represented by formula (AM-1) or formula (AM-2) (including the sub-formulas of each formula) or their salts can form solvates with solvents such as water, ethanol, and glycerol.

[0803] 4. Michael addition reaction

[0804] The Michael addition reaction refers to a reaction in which an electron-deficient double bond conjugated with an electron-withdrawing group (for example, acryloyl group, cinnamic acid group, maleimide group, etc.) undergoes a 1,4-addition (conjugate addition) reaction with a carbanion, an organometallic compound, an amine, an alcoholate, or a thiolate to form a covalent bond, as shown in the following reaction formula.

[0805] [Chemical formula 102]

[0806]

[0807] (In the aforementioned reaction formula, EWG represents an electron-withdrawing group (for example, COOR, CONHR, etc.); Nu - represents a nucleophilic reagent selected from a carbanion (R-M), R-NH 2 (or R-NH - ), R-OH (or R-O - ), R-SH (or R-S - ), etc.; R represents various substituents such as C 1～6 alkyl; R x or R y represents a hydrogen atom, various substituents such as C 1～6 alkyl; M represents a metal such as Li, Na).

[0808] More specifically, the Michael addition reaction in this specification is a reaction in which a maleimide compound shown in the following reaction formula undergoes a 1,4-addition (conjugate addition) reaction with a thiol compound to form a covalent bond.

[0809] [Chemical formula 103]

[0810]

[0811] (In the aforementioned reaction formula, R represents C1～6 (such as alkyl and other various substituents).

[0812] In some ways, the Michael addition reaction can form covalent bond-based crosslinks between alginic acid molecules without generating undesired by-products, in a short time, easily, and efficiently.

[0813] In the crosslinking method of alginic acid derivatives in a preferred way, almost no undesired by-products are formed in this Michael addition reaction. At this time, in the production of a new type of biocompatible material using alginic acid and the formation of an alginic acid hydrogel, various bioactive molecules can be introduced, and cell substances can be introduced through the alginic acid hydrogel for reconstructive surgery or for gene therapy.

[0814] 5. Crosslinked Alginate

[0815] There are (i) crosslinked alginate obtained by bonding through divalent metal ions, (ii) crosslinked alginate obtained by chemical bonding, or (iii) crosslinked alginate obtained by both divalent metal ion bonding and chemical bonding. Any crosslinked alginate has the property of forming a semi-solid from a gel-like state and sometimes forming a sponge-like morphology.

[0816] The crosslinked alginate obtained by bonding through divalent metal ions reacts at a super-high speed and is reversible. In contrast, the crosslinked alginate obtained by chemical bonding reacts slowly under relatively mild conditions and is irreversible. The physical properties of the crosslinked alginate can be adjusted by methods such as changing the concentration of the aqueous solution containing divalent metal ions (e.g., calcium chloride aqueous solution) used or the introduction rate of the reactive groups introduced into the alginic acid.

[0817] By utilizing the aforementioned crosslinking reaction, various alginate structures can be fabricated. For example, by using an ion crosslinking reaction, a specific structure can be instantaneously formed from an alginic acid solution. For the structural reinforcement of this structure (such as obtaining long-term stability, etc.), a crosslinking reaction based on chemical bonding can be utilized. Moreover, for example, in a crosslinked alginate structure obtained by both divalent metal ion bonding and chemical bonding, the divalent metal ions obtained by ion crosslinking are reversibly released, and a structure with only chemical bonding-based crosslinks remaining can also be fabricated.

[0818] A certain crosslinked alginate can be obtained by mixing the alginic acid derivatives of the aforementioned formula (I) and the aforementioned formula (II) and performing a Michael addition reaction.

[0819] In this specification, the implementation of the crosslinking reaction means: by performing a Michael addition reaction using the alginic acid derivative represented by the aforementioned formula (I) and the alginic acid derivative represented by the aforementioned formula (II), a chemical crosslinking (chemical bonding) is formed between the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II), or by co-existing the alginic acid derivative represented by the aforementioned formula (I) and the alginic acid derivative represented by the aforementioned formula (II) with a divalent metal ion, an ionic crosslinking (ionic bonding) is formed between the respective derivatives of the alginic acid derivative represented by formula (I) and / or the alginic acid derivative represented by formula (II), or both a chemical crosslinking based on the aforementioned Michael addition reaction and an ionic crosslinking based on the divalent metal ion are formed.

[0820] In this specification, it is possible to add a deprotecting agent (for example, when P 1 is not a hydrogen atom) in a mixed solution containing the alginic acid derivative of formula (I) and the alginic acid derivative of formula (II-P) (wherein, in formula (II-P), P 1 is not a hydrogen atom) or in a solution of a composition containing the alginic acid derivative of formula (I) and the alginic acid derivative of formula (II-P) (wherein, in formula (II-P), P 1 is not a hydrogen atom) in order to deprotect the protecting group P of the alginic acid derivative of the aforementioned formula (II-P). 1 When P is an acyl-based protecting group such as an acetyl group or a benzoyl group, an alkali such as an aqueous sodium hydroxide solution can be cited. The deprotecting agent can be appropriately selected according to the protecting group P 1 ), thereby forming a chemical crosslinking (chemical bonding).

[0821] In a certain mode, the crosslinked alginic acid forms a three-dimensional network structure through chemical crosslinking (covalent bond-based crosslinking formed by a maleimide group and a thiol group). Preferred alginic acid derivatives are substances with improved stability of the crosslinked alginic acid after crosslinking.

[0822] Some modes of crosslinked alginic acid are crosslinked alginic acids obtained by amide bonding between any carboxyl group of the first alginic acid and any carboxyl group of the second alginic acid through the following formula (LK-1).:

[0823] [Chemical formula 104]

[0824]

[0825] [In formula (LK-1), the -CONH- and -NHCO- at both ends represent amide bonds obtained through any carboxyl group of alginic acid; -L 1 -, and -L 2 - are the same as the definitions in the aforementioned mode [1].]

[0826] In some embodiments, the mixing ratio of the alginic acid derivative of formula (I) to the alginic acid derivative of formula (II) in the preparation of crosslinked alginic acid, based on the weight ratio of the derivative of formula (I) to the derivative of formula (II), is, for example, 1 to 1.5:1, preferably 1.2 to 1.5:1 or 1 to 1.2:1, and more preferably 1:1.

[0827] In some embodiments, the mixing ratio of the alginic acid derivative of formula (II) to the alginic acid derivative of formula (I) in the preparation of crosslinked alginic acid, based on the weight ratio of the derivative of formula (II) to the derivative of formula (I), is, for example, 1 to 4.0:1, preferably 1.5 to 4.0:1 or 1.2 to 1.5:1 or 1 to 1.2:1, and more preferably 1:1.

[0828] In some embodiments, the mixing ratio of the alginic acid derivative of formula (I) to the alginic acid derivative of formula (II) in the preparation of crosslinked alginic acid is more preferably based on the ratio of the introduction rate (mol%) of the reactive groups of the alginic acid derivative of formula (I) to the alginic acid derivative of formula (II), for example, 1 to 1.5:1, preferably 1.2 to 1.5:1 or 1 to 1.2:1, and more preferably 1:1.

[0829] In some embodiments, the mixing ratio of the alginic acid derivative of formula (II) to the alginic acid derivative of formula (I) in the preparation of crosslinked alginic acid is more preferably based on the ratio of the introduction rate (mol%) of the reactive groups of the alginic acid derivative of formula (II) to the alginic acid derivative of formula (I), for example, 1 to 4.0:1, preferably 1.5 to 4.0:1 or 1.2 to 1.5:1 or 1 to 1.2:1, and more preferably 1:1.

[0830] It should be noted that in the aforementioned mixing ratio, the alginic acid derivative of formula (I) can be respectively replaced by the alginic acid derivative of formula (II), and the alginic acid derivative of formula (II) can be replaced by the derivative of formula (I).

[0831] For crosslinked alginic acid, it is not necessary for all carboxyl groups of the constituent units of alginic acid to have the crosslinking of the above formula (LK-1). The introduction rate (also referred to as the crosslinking rate) of the crosslinking shown by the above formula (LK-1) in crosslinked alginic acid is, for example, in the range of 0.1 to 80%, 0.3 to 60%, 0.5 to 30%, or 1.0 to 10%.

[0832] The concentration of the alginic acid derivative of formula (I) or formula (II) in the Michael addition reaction for obtaining crosslinked alginic acid is usually in the range of 1 to 500 mg / mL, preferably 5 to 100 mg / mL.

[0833] The reaction temperature of the Michael addition reaction is usually 4 to 60 °C of the external temperature, preferably 15 to 40 °C of the external temperature.

[0834] The stirring time for forming crosslinked alginic acid (hydrogel) is, for example, several seconds to 24 hours, several seconds to 12 hours, several seconds to 30 minutes, or several seconds to 10 minutes.

[0835] The reaction solvent or reaction solution used in the Michael addition reaction is not particularly limited, and examples thereof include tap water, pure water (e.g., distilled water, ion-exchanged water, RO water, RO-EDI water, etc.), ultrapure water, cell culture medium, phosphate-buffered saline (PBS), and physiological saline, etc., and ultrapure water is preferred.

[0836] Some forms of crosslinked alginic acid are crosslinked alginic acids that include chemical crosslinking based on covalent bonds formed by Michael addition reaction and ionic crosslinking partially formed by divalent metal ions (e.g., calcium ions, etc.) as crosslinking.

[0837] 6. Composition

[0838] A composition containing the alginic acid derivative represented by the foregoing formula (I) and the alginic acid derivative represented by the foregoing formula (II) can be provided. (ALG), -NHCO-, -L 1 - and -L 2 - are as described in the foregoing manner.

[0839] [Chemical formula 105]

[0840]

[0841] In some forms of the composition, the weight ratio of the alginic acid derivative of formula (I) to the alginic acid derivative of formula (II) (alginic acid derivative of formula (I): alginic acid derivative of formula (II)) is, for example, 1:1 to 1.5, preferably 1:1.2 to 1.5 or 1:1 to 1.2, and more preferably 1:1.

[0842] In some forms of the composition, the weight ratio of the alginic acid derivative of formula (II) to the alginic acid derivative of formula (I) (alginic acid derivative of formula (II): alginic acid derivative of formula (I)) is, for example, 1:1 to 1.5, preferably 1:1.2 to 1.5 or 1:1 to 1.2, and more preferably 1:1.

[0843] In some forms of the composition, the mixing ratio of the alginic acid derivative of formula (I) to the alginic acid derivative of formula (II) is based on the ratio of the introduction rate (mol%) of the group of formula (BR-1) in the alginic acid derivative of formula (I) to the group of formula (BR-2) in the alginic acid derivative of formula (II), and is, for example, 1:1 to 1.5, preferably 1:1.2 to 1.5 or 1:1 to 1.2, and more preferably 1:1.

[0844] In some embodiments of the composition, the mixing ratio of the alginic acid derivative of formula (II) to the alginic acid derivative of formula (I) is based on the ratio of the introduction rate (mol%) of the group of formula (BR-2) in the alginic acid derivative of formula (II) to the group of formula (BR-1) in the alginic acid derivative of formula (I), and is, for example, 1:1 to 1.5, preferably 1:1.2 to 1.5 or 1:1 to 1.2, more preferably 1:1.

[0845] There can be provided a composition comprising the alginic acid derivative represented by the aforementioned formula (I) and the alginic acid derivative represented by the aforementioned formula (II-P). (ALG), -NHCO-, P in each formula 1 , -L 1 -, and -L 2 - are as described in the aforementioned manner.

[0846] [Chemical formula 106]

[0847]

[0848] In some embodiments of the composition, the weight ratio of the alginic acid derivative of formula (I) to the alginic acid derivative of formula (II-P) (alginic acid derivative of formula (I): alginic acid derivative of formula (II-P)) is, for example, 1:1 to 1.5, preferably 1:1.2 to 1.5 or 1:1 to 1.2, more preferably 1:1.

[0849] In some embodiments of the composition, the weight ratio of the alginic acid derivative of formula (II-P) to the alginic acid derivative of formula (I) (alginic acid derivative of formula (II-P): alginic acid derivative of formula (I)) is, for example, 1:1 to 1.5, preferably 1:1.2 to 1.5 or 1:1 to 1.2, more preferably 1:1.

[0850] In some embodiments of the composition, the mixing ratio of the alginic acid derivative of formula (I) to the alginic acid derivative of formula (II-P) is based on the ratio of the introduction rate (mol%) of the group of formula (BR-1) in the alginic acid derivative of formula (I) to the group of formula (BR-2-P) in the alginic acid derivative of formula (II-P), and is, for example, 1:1 to 1.5, preferably 1:1.2 to 1.5 or 1:1 to 1.2, more preferably 1:1.

[0851] In some embodiments of the composition, the mixing ratio of the alginic acid derivative of formula (II-P) to the alginic acid derivative of formula (I) is based on the ratio of the introduction rate (mol%) of the group of formula (BR-2-P) in the alginic acid derivative of formula (II-P) to the group of formula (BR-1) in the alginic acid derivative of formula (I), and is, for example, 1:1 to 1.5, preferably 1:1.2 to 1.5 or 1:1 to 1.2, more preferably 1:1.

[0852] 7. Crosslinked alginic acid structure

[0853] The crosslinked alginic acid structure can be obtained by a method including performing a crosslinking reaction on the aforementioned alginic acid derivative. As specific crosslinked alginic acid structures, for example, fibrous structures, fibers, beads, gels, nearly spherical gels, etc. can be mentioned. It is preferable that the stability of the crosslinked alginic acid structure is improved. In addition, the crosslinked alginic acid structure may have the ability to hold a content inside it (content retention ability). The crosslinked alginic acid structure can be prepared, for example, by the following methods, but is not limited to them.

[0854] [Mixing method (1)]

[0855] By dropping a mixed solution of an alginic acid derivative of formula (I) and an alginic acid derivative of formula (II) or a solution of a composition containing the alginic acid derivative shown in formula (I) and the alginic acid derivative shown in formula (II) into a solution containing a divalent metal ion, a specific structure, namely a crosslinked alginic acid structure, which forms chemical crosslinking (covalent bond-based crosslinking formed by Michael addition reaction) and ionic crosslinking (crosslinking partially formed by divalent metal ions), can be obtained.

[0856] [Mixing method (2)]

[0857] In a mixed solution of an alginic acid derivative of formula (I) and an alginic acid derivative of formula (II-P) (wherein in formula (II-P), P 1 is not a hydrogen atom) or a solution of a composition containing the alginic acid derivative shown in formula (I) and the alginic acid derivative shown in formula (II-P) (wherein in formula (II-P), P 1 is not a hydrogen atom), in order to deprotect the protecting group P 1 of the alginic acid derivative of formula (II-P) above, a deprotecting agent is added (for example, when P 1 is an acyl-based protecting group such as an acetyl group or a benzoyl group, alkalis such as an aqueous sodium hydroxide solution can be mentioned. The deprotecting agent can be appropriately selected according to the protecting group P 1 ) and then dropped into a solution containing a divalent metal ion, whereby a specific structure, namely a crosslinked alginic acid structure, which forms chemical crosslinking (covalent bond-based crosslinking formed by Michael addition reaction) and ionic crosslinking (crosslinking partially formed by divalent metal ions), can be obtained.

[0858] [Coating method (1)]

[0859] A solution of the alginic acid derivative of formula (I) is dropped into a solution containing a divalent metal ion or the like to obtain a specific structure that is partially crosslinked. By adding the structure obtained above, such as a gel, into the solution of the alginic acid derivative of formula (II), and further carrying out a crosslinking reaction (Michael addition reaction) on the surface of the structure, etc., a crosslinked alginic acid structure can be obtained. It should be noted that this method can also be carried out by replacing the alginic acid derivative of formula (I) with the alginic acid derivative of formula (II) and replacing the alginic acid derivative of formula (II) with the alginic acid derivative of formula (I) respectively.

[0860] [Coating method (2)]

[0861] A solution of the alginic acid derivative of formula (I) is dropped into a solution containing a divalent metal ion or the like to obtain a specific structure that is partially crosslinked. By adding the structure obtained above, such as a gel, into the solution of the alginic acid derivative of formula (II-P) (wherein in formula (II-P), P 1 is not a hydrogen atom), and further adding a deprotecting agent in order to deprotect the protecting group P 1 of the alginic acid derivative represented by the above formula (II-P) (for example, when P 1 is an acyl-based protecting group such as an acetyl group or a benzoyl group, an alkali such as an aqueous sodium hydroxide solution can be cited. The deprotecting agent can be appropriately selected according to the protecting group P 1 ), thereby further carrying out a crosslinking reaction (Michael addition reaction) on the surface of the structure, etc., and a crosslinked alginic acid structure can be formed.

[0862] And, a solution of the alginic acid derivative of formula (II-P) (wherein in formula (II-P), P 1 is not a hydrogen atom) is dropped into a solution containing a divalent metal ion or the like to obtain a specific structure that is partially crosslinked. By adding the structure obtained above, such as a gel, into the solution of the alginic acid derivative of formula (I), and further adding a deprotecting agent in order to deprotect the protecting group P 1 of the alginic acid derivative represented by the above formula (II-P) (for example, when P 1 is an acyl-based protecting group such as an acetyl group or a benzoyl group, an alkali such as an aqueous sodium hydroxide solution can be cited. The deprotecting agent can be appropriately selected according to the protecting group P 1 ), thereby further carrying out a crosslinking reaction (Michael addition reaction) on the surface of the structure, etc., and a crosslinked alginic acid structure can be formed.

[0863] The divalent metal ion used in the above method is not particularly limited. For example, calcium ion, magnesium ion, barium ion, strontium ion, zinc ion, etc. can be cited, and calcium ion is preferred.

[0864] The calcium ion-containing solution used in the aforementioned method is not particularly limited. For example, aqueous solutions such as calcium chloride aqueous solution, calcium carbonate aqueous solution, and calcium gluconate aqueous solution can be cited, and a calcium chloride aqueous solution is preferred.

[0865] The calcium ion concentration of the calcium ion-containing solution used in the aforementioned method is not particularly limited. For example, 1 mM to 1 M can be cited, preferably 5 mM to 500 mM, and more preferably 10 mM to 300 mM.

[0866] The solvent or solution used in the aforementioned method is also not particularly limited. For example, tap water, pure water (e.g., distilled water, ion-exchanged water, RO water, RO-EDI water, etc.), ultrapure water, cell culture medium, phosphate-buffered saline (PBS), and physiological saline, etc. can be cited, and ultrapure water is preferred.

[0867] The physical properties of the alginate gel can be adjusted by physical property values such as hardness, elasticity, repulsive force, breaking force, and stress at rupture.

[0868] 8. Biocompatibility of Alginate Derivatives, Crosslinked Alginate, and Crosslinked Alginate Structures In this specification, alginate derivatives, crosslinked alginate, and crosslinked alginate structures have biocompatibility. In this specification, biocompatibility refers to the property of not causing reactions such as the interaction between a biomaterial (here, referring to an alginate derivative represented by formula (I) or formula (II), and a crosslinked alginate or crosslinked alginate structure prepared using these two alginate derivatives) and a living body, the local reaction of the tissue adjacent to the aforementioned biomaterial, or the systemic reaction, etc., and is called having biocompatibility.

[0869] In this specification, regarding the biocompatibility of alginate derivatives, crosslinked alginate, or crosslinked alginate structures, it can be confirmed by the examples of biocompatibility described later.

[0870] 9. Stability of Crosslinked Alginate Structures

[0871] The stability of the crosslinked alginate structure can be confirmed by, for example, measuring the gel stability, and the permeability can be confirmed by measuring the gel permeability, etc.

[0872] [Method for Measuring Gel Stability]

[0873] Add phosphate buffered saline (PBS) to the crosslinked alginate structure gel in a container, and measure the concentration (μg / mL) of alginic acid leaked into the PBS. The value obtained by dividing the measured alginic acid concentration by the total alginic acid concentration obtained by decomposing the crosslinked alginate structure gel and expressed as a percentage is defined as the disintegration rate. Specifically, the gel stability can be determined by the method described in the following examples.

[0874] In this specification, the gel disintegration rate of the crosslinked alginate structure is preferably 0% to 90%, more preferably 0% to 70%, and even more preferably 0% to 50%. The stability of the crosslinked alginate structure means that the lower the concentration of alginic acid leaked into the aqueous solution, that is, the lower the gel disintegration rate, the higher the stability.

[0875] [Method for measuring gel permeability]

[0876] Prepare a crosslinked alginate structure gel encapsulating fluorescein isothiocyanate-dextran, add physiological saline to the aforementioned gel in a container, and measure the concentration of dextran leaked into the physiological saline. The value obtained by dividing the measured concentration of dextran by the total dextran concentration obtained by decomposing the crosslinked alginate structure gel encapsulating fluorescein isothiocyanate-dextran and expressed as a percentage is the gel permeability. Specifically, the gel permeability can be determined by the method described in the following examples.

[0877] For the gel permeability of crosslinked alginic acid after 24 hours of adding physiological saline, for example, when encapsulating dextran with a molecular weight of 2 million, it is preferably 0% to 90%, more preferably 0% to 70%, and even more preferably 0% to 50%. In addition, when encapsulating dextran with a molecular weight of 150,000, for example, if the use purpose of the crosslinked alginate structure gel is the release / generation of proteins and antibodies, it is preferably 1% to 100%, more preferably 10% to 100%, and even more preferably 30% to 100%. In addition, if the use purpose is an immune barrier, it is preferably 0% to 90%, more preferably 0% to 70%, and even more preferably 0% to 50%.

[0878] Regarding the permeability of the crosslinked alginate structure, the lower the permeability, the lower the permeability of the content and substances outside the gel, and the higher the permeability, the higher the permeability of the content and substances outside the gel.

[0879] The permeability of the gel can be adjusted according to the molecular weight, concentration of the alginic acid used, the type of crosslinking group introduced into the alginic acid, the introduction rate, the type and concentration of the divalent metal ions used in gelation, or a combination thereof.

[0880] [Method for preparing a crosslinked alginate structure gel encapsulating a content]

[0881] For example, a crosslinked alginate structure gel encapsulating fluorescein isothiocyanate-dextran as a content can be prepared by the following method.

[0882] (1) Mix a solution of the alginate derivative shown in formula (I) and a fluorescein isothiocyanate-dextran solution.

[0883] (2) Mix a solution of the alginate derivative shown in formula (II) in the mixed solution obtained in (1).

[0884] (When formula (I) in (1) is changed to formula (II), formula (II) in (2) is changed to formula (I))

[0885] (3) A gel obtained by dropping the mixed solution obtained in (2) into a solution containing calcium ions forms chemical crosslinking and ionic crosslinking in the solution, whereby a crosslinked alginate structure gel encapsulating fluorescein isothiocyanate-dextran can be obtained.

[0886] 10. Uses of alginate derivatives and crosslinked alginate structures

[0887] Alginate derivatives can be used instead of conventional alginate in a wide range of fields such as food, medicine, cosmetics, fibers, and papermaking. As preferred uses of alginate derivatives or crosslinked alginate structures, specifically, medical materials such as wound dressing materials, postoperative anti-adhesion materials, drug sustained-release substrates, cell culture substrates, and cell transplantation substrates can be cited. As diseases to be treated based on cell transplantation, diabetes, Parkinson's disease, hemophilia, etc. can be cited.

[0888] As the shape of the crosslinked alginate structure when used as a medical material, fibrous structures, fibers, beads, gels, nearly spherical gels, etc. can be cited, preferably beads, gels, or nearly spherical gels, and more preferably nearly spherical gels.

[0889] It should be noted that all publications cited in this specification, such as prior art documents, published gazettes, patent gazettes, and other patent documents, are incorporated into this specification by reference.

[0890] Moreover, the objects, features, advantages, and concepts of the present invention are obvious to those skilled in the art from the description of this specification. According to the description of this specification, those skilled in the art can easily implement the present invention. The best mode for carrying out the invention and specific examples, etc. represent preferred embodiments of the present invention and are shown for illustration or explanation, and the present invention is not limited to them. It is obvious to those skilled in the art that various modifications can be made within the spirit and scope of the present invention disclosed in this specification based on the description of this specification.

[0891] Examples

[0892] The measurement of nuclear magnetic resonance spectroscopy (NMR) was performed using a JEOL JNM-ECX400 FT-NMR (JEOL, Japan). Liquid chromatography-mass spectrometry (LC-Mass) was measured using the following method. Using a Waters AQUITY UPLC system and a BEH C18 column (2.1 mm × 50 mm, 1.7 μm) (Waters), the mobile phase and gradient conditions of acetonitrile: 0.05% trifluoroacetic acid aqueous solution = 5:95 (0 min) - 95:5 (1.0 min) - 95:5 (1.6 min) - 5:95 (2.0 min) were used.

[0893] 1 In the 1H-NMR data, in the NMR signal pattern, s represents singlet, d represents doublet, t represents triplet, q represents quartet, m represents multiplet, br represents broad range, J represents coupling constant, Hz represents Hertz, CDCl 3 represents deuterated chloroform, DMSO-d 6 represents deuterated dimethyl sulfoxide, D 2 O represents deuterated water, CD 3 OD represents deuterated methanol. 1 In the 1H-NMR data, regarding the signals of hydroxyl group (OH), amino group (NH 2 )), carboxyl group (COOH) protons, etc. that could not be confirmed due to being broad bands, the data was not recorded. In the LC-Mass data, M represents molecular weight, and [M+H]+ represents the molecular ion peak.

[0894] "Room temperature" in the examples generally refers to a temperature of about 0 °C to about 35 °C.

[0895] The "introduction rate" in the examples was determined by performing 1H-NMR measurement in D 2 O, and was recorded as "mol% (NMR integration ratio)" based on the ratio of the proton integration values of the maleimide group or aromatic ring of the reactive substituent to the alginic acid.

[0896] (Example 1)

[0897] Synthesis of alginic acid (AL-EX-1) incorporating 2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethylamino

[0898] [Chemical formula 107]

[0899]

[0900] <Step 1>

[0901] Synthesis of tert-Butyl (2-(2,5-Dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)Carbamate (EX1-IM-1) Maleic anhydride (600 mg) was suspended in ethanol (6.0 mL), and a solution of tert-butyl (2-aminoethyl)carbamate (1.03 g) and triethylamine (0.90 mL) in ethanol (3.0 mL) was added under ice-water cooling. After the reaction mixture was stirred at room temperature for 2 hours, ethanol was distilled off under reduced pressure. The residue was dissolved in acetic anhydride (6.0 mL), sodium acetate (502 mg) was added, and the mixture was stirred at 70 °C for 1.5 hours. Ethyl acetate (25 mL) and water (10 mL) were added, and the layers were separated. The organic layer was washed successively with saturated aqueous sodium bicarbonate (10 mL, 3 times) and saturated brine (10 mL), dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (heptane to 50% ethyl acetate / heptane). The resulting oil was triturated with heptane (20 mL). The solid was collected by filtration, washed with heptane, and dried under reduced pressure to obtain the title compound (1.01 g) as a white solid.

[0902] <Step 2>

[0903] Synthesis of 1-(2-Aminoethyl)-1H-pyrrole-2,5-dione Hydrochloride (EX1-IM-2) To the compound (500 mg) obtained in <Step 1> of (Example 1) was added 4 N hydrochloric acid in ethyl acetate solution (5.0 mL), and the mixture was stirred at room temperature for 1.5 hours. After adding ethyl acetate (5.0 mL), the precipitate was collected by filtration and washed with ethyl acetate. The resulting hygroscopic solid was suspended in ethyl acetate, and ethyl acetate was distilled off under reduced pressure and then dried under reduced pressure to obtain the title compound (328 mg) as a white solid.

[0904] <Step 3>

[0905] Synthesis of Alginate (AL-EX-1) Incorporating 2-(2,5-Dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl Amine To an aqueous solution (20 mL) of sodium alginate (manufactured by Kimika Co., Ltd., ALG-2) prepared to be 1 wt% was added the compound (36 mg) obtained in <Step 2> of (Example 1), 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (84 mg), and 1 M aqueous sodium bicarbonate (252 μL), and the mixture was stirred at 30 °C for 3 hours. After adding sodium chloride (200 mg), ethanol (40 mL) was added, and the mixture was stirred at room temperature for 30 minutes. The resulting precipitate was collected by filtration, washed with ethanol, and dried under reduced pressure to obtain the title compound (183 mg) as a white solid.

[0906] (Example 2)

[0907] (Example 2)

[0908] Synthesis of alginic acid (AL-EX-2) incorporating 2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)ethylamino [Chemical 108]

[0909]

[0910] <Step 1>

[0911] Synthesis of tert-butyl (2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)ethyl)carbamate (EX2-IM-1) 1H-Pyrrole-2,5-dione (0.7 g), tert-butyl (2-(2-hydroxyethoxy)ethyl)carbamate (1.0 g), and triphenylphosphine (1.4 g) were dissolved in tetrahydrofuran (20 mL). Diisopropyl azodicarboxylate (1.9 mol / L - toluene solution, 2.8 mL) was added dropwise under cooling with ice-salt water, and then the mixture was stirred for 30 minutes under ice-water cooling. After stirring for 1 hour at room temperature, ethyl acetate (20 mL) and water (10 mL) were added, and the layers were separated. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (heptane - ethyl acetate) and then dried under reduced pressure to obtain the title compound (0.5 g) as a pale yellow oil.

[0912] <Step 2>

[0913] Synthesis of 1-(2-(2-aminoethoxy)ethyl)-1H-pyrrole-2,5-dione trifluoroacetate (EX2-IM-2) Trifluoroacetic acid (2.3 mL) was added to the compound (0.5 g) obtained in <Step 1> of (Example 2) under ice-water cooling, and the mixture was stirred for 1 hour at room temperature. Diisopropyl ether (11.3 mL) was added, and after stirring for 30 minutes at room temperature, the precipitated solid was filtered off and washed with diisopropyl ether. The resulting hygroscopic solid was suspended in diisopropyl ether, the solvent was distilled off, and then dried under reduced pressure to obtain the title compound (0.3 g) as a pale yellow solid.

[0914] <Step 3>

[0915] Synthesis of alginic acid (AL-EX-2) incorporating 2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)ethylamino Using an aqueous solution (20 mL) of sodium alginate (manufactured by Kimika Co., Ltd., ALG-2) prepared to be 1 wt% and the compound (60 mg) obtained in <Step 2> of (Example 2), the same operation as <Step 3> of (Example 1) was carried out to obtain the title compound (183 mg) as a white solid.

[0916] (Example 3)

[0917] Synthesis of Alginate (AL-EX-3) Incorporating 2-(2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)ethoxy)ethylamino

[0918] [Chemical 109]

[0919]

[0920] <Step 1>

[0921] (Synthesis of tert-butyl (2-(2-(2-aminoethoxy)ethoxy)ethyl)carbamate (EX3-IM-1)) A solution of di-tert-butyl dicarbonate (3.0 g) in dichloromethane (37.5 mL) was added dropwise to a solution of 2,2’-(ethane-1,2-diylbis(oxy))ethane-1-amine (3.2 g) and triethylamine (11.5 mL) in dichloromethane (30.0 mL) over 4.75 hours while cooling with ice-water. The mixture was then stirred at room temperature for 18.5 hours. The reaction mixture was concentrated under reduced pressure. Dichloromethane (30 mL) was added to the residue, and the insoluble material was filtered off. The filtrate was washed successively with water (10 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was dried under reduced pressure to give the title crude compound (2.7 g) as a colorless oil.

[0922] <Step 2>

[0923] (Synthesis of tert-butyl (2-(2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)ethoxy)ethyl)carbamate (EX3-IM-2)) The compound obtained in <Step 1> of (Example 3) (500 mg) and maleic anhydride (217 mg) were suspended in ethanol (5.0 mL) and stirred at room temperature for 30 minutes. Ethanol was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (heptane - ethyl acetate) to give the amide (423 mg). Acetic anhydride (4.2 mL) was added to the resulting colorless oil and sodium acetate (100 mg), and the mixture was stirred at 40 °C for 1 hour, then at 60 °C for 1 hour, at 80 °C for 1.5 hours, and at 100 °C for 2 hours. Ethyl acetate (25 mL) and water (10 mL) were added to the reaction mixture, and the layers were separated. The organic layer was washed successively with saturated aqueous sodium hydrogen carbonate (10 mL), water (10 mL), and saturated brine (5 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (heptane - 80% ethyl acetate / heptane) to give the title compound (275 mg) as a colorless oil.

[0924] <Step 3>

[0925] Synthesis of 1-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-1H-pyrrole-2,5-dione trifluoroacetate (EX3-IM-3): Under ice-water cooling, trifluoroacetic acid (1.9 mL) was added to the compound (275 mg) obtained in <Step 2> of (Example 3), and the mixture was stirred at room temperature for 15 minutes. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (ethyl acetate to 30% methanol / ethyl acetate) to obtain the title compound (231 mg) as a colorless oil.

[0926] <Step 4>

[0927] Synthesis of alginic acid (AL-EX-3) incorporating 2-(2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)ethoxy)ethylamino

[0928] Using an aqueous solution (20 mL) of sodium alginate (manufactured by Kimica Corporation, ALG-2) prepared at 1 wt% and the compound (69 mg) obtained in <Step 3> of (Example 3), the same operation as <Step 3> of (Example 1) was carried out to obtain the title compound (145 mg) as a white solid.

[0929] (Example 4)

[0930] Synthesis of alginic acid (AL-EX-4) incorporating 2-amino-N-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)acetamido

[0931] [Chemical formula 110]

[0932]

[0933] <Step 1>

[0934] (2-((2-(2,5-Dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)amino)-2-oxoethyl)carbamic acid tert-butyl ester (EX4-IM-1) was synthesized. To a mixture of commercially available 1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride [CAS No. 134272-64-3] (92.43 mg) and water (750 μL) was added 1 molar concentration-sodium bicarbonate water (578.5 μL) at room temperature. To this mixture was added a solution of commercially available 2,5-dioxopyrrolidin-1-yl (tert-butoxycarbonyl)glycinate [CAS: 3392-07-2] (150 mg) in tetrahydrofuran (1500 μL) at room temperature, and the mixture was stirred at the same temperature for 30 minutes. After completion of the reaction, ethyl acetate (10 mL) and water (5 mL) were added, and separation was carried out. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (25% ethyl acetate / heptane to 100% ethyl acetate, ethyl acetate to 60% methanol / ethyl acetate) to obtain the title compound (74 mg) as a colorless oily compound.

[0935] <Step 2>

[0936] Synthesis of 2-amino-N-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)acetamide trifluoroacetate (EX4-IM-2). To a mixture of the compound (0.074 g) obtained in <Step 1> of (Example 4) and dichloromethane (0.22 mL) was added trifluoroacetic acid (0.52 mL) with stirring under ice cooling, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was concentrated, and diisopropyl ether (20 mL) was added. Since a colloidal compound was formed, the mixture was concentrated under reduced pressure and dried to obtain the title crude compound (0.097 g) as a pale yellow colloidal compound.

[0937] <Step 3>

[0938] Synthesis of alginic acid (AL-EX-4) incorporating 2-amino-N-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)acetamido. At room temperature, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (68.6 mg) and 1 molar concentration sodium bicarbonate solution (68.6 μL) were added to an aqueous solution (29.7 mL) of sodium alginate (manufactured by Kimika Co., Ltd., ALG-2) prepared at 1 wt%. Subsequently, a mixture of the compound obtained in <Step 2> of Example 4 (21.4 mg), water (1 mL), and ethanol (1 mL) was slowly added at the same temperature, and the mixture was stirred at 40 °C for 4 hours. After adding sodium chloride (300 mg), ethanol (59.3 mL) was added, and the mixture was stirred at room temperature for 30 minutes. The resulting precipitate was collected by filtration, washed with ethanol, and dried under reduced pressure to obtain the title compound (221.3 mg) in the form of a white flocculent compound.

[0939] (Example 5)

[0940] Synthesis of alginic acid (AL-EX-5) incorporating (S)-2-amino-N-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)-3-phenylpropanamido

[0941] [Chemical Formula 111]

[0942]

[0943] <Step 1>

[0944] Synthesis of tert-butyl (S)-(1-((2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)amino)-1-oxo-3-phenylpropan-2-yl)carbamate (EX5-IM-1). To a mixture of commercially available 1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride [CAS: 134272-64-3] (100 mg), commercially available (tert-butoxycarbonyl)-L-phenylalanine [CAS No. 13734-34-4] (150.23 mg), and dichloromethane (1 mL), triethylamine (78.9 μL) was added with stirring under ice-cooling. To this mixture, N,N'-dicyclohexylcarbodiimide (116.8 mg) was added at the same temperature, and the mixture was stirred at room temperature for 30 minutes. After completion of the reaction, the mixture was diluted with ethyl acetate (20 mL), and the suspension was filtered. The crude product was purified by silica gel column chromatography (12% ethyl acetate / heptane to 100% ethyl acetate) to obtain the title compound (108 mg) in the form of a white amorphous solid.

[0945] <Step 2>

[0946] (S)-2-Amino-N-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)-3-phenylpropanamide trifluoroacetate (EX5-IM-2) Synthesis To a mixture of the compound obtained in <Step 1> of Example 5 (0.1 g) and dichloromethane (1.3 mL), trifluoroacetic acid (0.7 mL) was added under stirring with ice cooling, and the mixture was stirred at room temperature for 30 minutes. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and diisopropyl ether (20 mL) was added. The suspension was filtered to obtain the title compound (0.12 g) as a white solid.

[0947] <Step 3>

[0948] Synthesis of Alginate (AL-EX-5) Incorporating (S)-2-Amino-N-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)-3-phenylpropanamido Using an aqueous solution (29.7 mL) of sodium alginate (manufactured by Kimika Co., Ltd., ALG-2) prepared to be 1 wt% and the compound obtained in <Step 2> of Example 5 (27.5 mg), the same operation as <Step 3> of Example 4 was carried out to obtain the title compound (264.8 mg) as a white flocculent compound.

[0949] (Example 6)

[0950] Synthesis of Alginate (AL-EX-6) Incorporating (S)-2-(2-Aminoacetamido)-N-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)-3-phenylpropanamido

[0951] [Chemical Formula 112]

[0952]

[0953] <Step 1>

[0954] Synthesis of (tert-Butoxycarbonyl)glycyl-L-phenylalanine (EX6-IM-1)

[0955] To a mixture of commercially available L-phenylalanine [CAS: 63-91-2] (0.12 g) and water (1 mL), 1 molar concentration - sodium bicarbonate solution (0.73 mL) was added at room temperature. To this mixture, a solution of commercially available 2,5-dioxopyrrolidin-1-yl (tert-butoxycarbonyl) glycinate [CAS: 3392-07-2] (0.2 g) in tetrahydrofuran (4 mL) was added at room temperature, and the mixture was stirred at the same temperature. After 1 hour and 30 minutes, 2,5-dioxopyrrolidin-1-yl (tert-butoxycarbonyl) glycinate (0.02 g) was further added, and the mixture was stirred at room temperature for 30 minutes. After completion of the reaction, ethyl acetate (10 mL) and 1 normal concentration - hydrochloric acid (3 mL) were added, and the layers were separated. The organic layer was washed successively with water (5 mL) and saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (25% ethyl acetate / heptane to 100% ethyl acetate, ethyl acetate to 20% methanol / ethyl acetate) to obtain the title compound (0.21 g) as a white amorphous solid.

[0956] <Step 2>

[0957] (S)-(2-((1-((2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)amino)-1-oxo-3-phenylpropan-2-yl)amino)-2-oxoethyl) carbamate (EX6-IM-2) was synthesized. To a mixture of commercially available 1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride [CAS: 134272-64-3] (114 mg), the compound obtained in <Step 1> of Example 6 (208 mg), and dichloromethane (2080 μL), triethylamine (90 μL) was added with stirring under ice-cooling. To this mixture, N,N'-dicyclohexylcarbodiimide (133.1 mg) was added at the same temperature, and the mixture was stirred at room temperature for 1 hour and 30 minutes. After completion of the reaction, the mixture was diluted with ethyl acetate (20 mL), and the suspension was filtered. The crude product was purified by silica gel column chromatography (25% ethyl acetate / heptane to 100% ethyl acetate, ethyl acetate to 20% methanol / ethyl acetate). The recovered fractions were concentrated under reduced pressure and dissolved in tert-butyl methyl ether (20 mL). This solution was washed successively with saturated sodium bicarbonate solution (5 mL), water (5 mL) twice, and saturated brine (5 mL), and dried over anhydrous sodium sulfate. The organic layer was concentrated under reduced pressure to obtain the title compound (220 mg) as a white amorphous solid.

[0958] <Step 3>

[0959] (S)-2-(2-Aminoacetamido)-N-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)-3-phenylpropanamide trifluoroacetate (EX6-IM-3) was synthesized using the compound (0.22 g) obtained in <Step 2> of (Example 6), and the same operation as <Step 2> of (Example 5) was carried out to obtain the title compound (0.25 g) in the form of a white solid.

[0960] <Step 4>

[0961] Synthesis of alginic acid (AL-EX-6) incorporating (S)-2-(2-aminoacetamido)-N-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)-3-phenylpropanamido Using an aqueous solution (49.4 mL) of sodium alginate (manufactured by Kimika Co., Ltd., ALG-2) prepared to 1 wt% and the compound (52.4 mg) obtained in <Step 3> of (Example 6), the same operation as <Step 3> of (Example 4) was carried out to obtain the title compound (485 mg) in the form of a white flocculent compound.

[0962] (Example 7)

[0963] Synthesis of alginic acid (AL-EX-7) incorporating S-(4-(2-aminoethyl)carbamoyl)benzyl)thioacetate

[0964] [Chemical Formula 113]

[0965]

[0966] <Step 1>

[0967] Synthesis of tert-butyl (2-(4-(chloromethyl)benzamido)ethyl)carbamate (EX7-IM-1) 4-(Chloromethyl)benzoyl chloride (2.0 g) was dissolved in tetrahydrofuran (10.0 mL), and a solution of tert-butyl (2-aminoethyl)carbamate (1.7 g) and diisopropylethylamine (3.7 mL) in tetrahydrofuran (10.0 mL) was added dropwise under ice-water cooling, and the mixture was stirred at room temperature for 1.5 hours. Ethyl acetate (30 mL) and water (10 mL) were added to the reaction solution, and the layers were separated. The organic layer was washed successively with semi-saturated aqueous sodium bicarbonate (10 mL), water (10 mL), and saturated brine (5 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was triturated with tert-butyl methyl ether, and the resulting solid was collected by filtration and washed with tert-butyl methyl ether to obtain the title compound (2.9 g) in the form of a white solid.

[0968] <Step 2>

[0969] Synthesis of S-(4-((2-((tert-Butoxycarbonyl)amino)ethyl)carbamoyl)benzyl)thioacetate (EX7-IM-2) The compound obtained in <Step 1> of (Example 7) (1.20 g) was suspended in acetonitrile (24.0 mL). Potassium thioacetate (0.53 g) was added, and the mixture was stirred at room temperature for 30 minutes. Ethyl acetate (50 mL) and water (20 mL) were added to the reaction solution, and the layers were separated. The organic layer was washed successively with water (20 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was triturated with tert-butyl methyl ether, and the solid was collected by filtration and washed with tert-butyl methyl ether. The obtained solid was dried under reduced pressure at 40 °C to give the title compound (1.27 g) as a white solid.

[0970] <Step 3>

[0971] Synthesis of S-(4-((2-Aminoethyl)carbamoyl)benzyl)thioacetate hydrochloride (EX7-IM-3) To the compound obtained in <Step 2> of (Example 7) (0.60 g) under ice-water cooling was added 4 N hydrochloric acid / 1,4-dioxane (4.2 mL), and the mixture was stirred at room temperature for 30 minutes. 4 N hydrochloric acid / 1,4-dioxane (2.1 mL) was added, and the mixture was further stirred at room temperature for 30 minutes. Diisopropyl ether (12.6 mL) was added to the reaction solution, and the resulting precipitate was collected by filtration, washed with diisopropyl ether, and dried under reduced pressure to give the title compound (0.46 g) as a white solid.

[0972] <Step 4>

[0973] Synthesis of Alginate incorporating S-(4-((2-Aminoethyl)carbamoyl)benzyl)thioacetate group (AL-EX-7) Using an aqueous solution of sodium alginate (manufactured by Kimica Corporation, ALG-2) prepared to be 1 wt% (20 mL) and the compound obtained in <Step 3> of (Example 7) (58 mg), the same operation as <Step 3> of (Example 1) was carried out to give the title compound (189 mg) as a white solid.

[0974] (Example 7.1)

[0975] Preparation of Alginate incorporating 2-(N-(4-(Mercaptomethyl)benzamido))ethylamino (AL-EX-7.1)

[0976] [Chemical formula 114]

[0977]

[0978] Dissolve the compound (160 mg) obtained in <Step 4> of (Example 7) in water (8.0 mL), add 1 N aqueous sodium hydroxide solution (112 μL), stir at 25 °C for 2 hours to prepare a 2 wt% solution of the title compound. When ethanol precipitation treatment was carried out, it formed a gel-like substance, so the solution was directly used for the test. Ethanol treatment was partially carried out, and the disappearance of the acetyl group was confirmed by NMR.

[0979] (Example 8)

[0980] Synthesis of alginic acid (AL-EX-8) incorporating S-(4-(3-((3-aminopropyl)amino)-3-oxopropyl)benzyl)thioacetate

[0981] [Chemical Formula 115]

[0982]

[0983] <Step 1>

[0984] Synthesis of methyl 4-(3-((3-((tert-butoxycarbonyl)amino)propyl)amino)-3-oxopropyl)benzoate (EX8-IM-1) Dissolve 3-(4-(methoxycarbonyl)phenyl)propanoic acid (1.15 g) and tert-butyl (3-aminopropyl)carbamate (0.96 g) in methanol (11.5 mL). Add 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (2.14 g), stir at room temperature for 2 hours and at 40 °C for 1 hour. Add ethyl acetate (20 mL) and water (20 mL) to the reaction solution, separate the layers, and extract the aqueous layer with ethyl acetate (10 mL). Combine the organic layers, wash successively with semi-saturated sodium bicarbonate solution (10 mL), water (10 mL), and saturated brine (5 mL), dry over anhydrous sodium sulfate, and distill off the solvent under reduced pressure. Purify the residue by silica gel column chromatography (10% ethyl acetate / heptane to ethyl acetate) to obtain the title compound (0.76 g) as a colorless oil.

[0985] <Step 2>

[0986] Synthesis of tert-butyl (3-(3-(4-(hydroxymethyl)phenyl)propanamido)propyl)carbamate (EX8-IM-2) Dissolve the compound (560 mg) obtained in <Step 1> of (Example 8) in tetrahydrofuran (11.2 mL). Add lithium aluminum hydride (146 mg) over 5 minutes and stir at room temperature for 1 hour. Under ice-water cooling, add saturated aqueous sodium sulfate solution (50 drops), and stir at the same temperature for 1 hour. Filter off the precipitated insoluble matter and wash with tetrahydrofuran. Concentrate the filtrate under reduced pressure to obtain the title compound (569 mg) as a colorless oil.

[0987] <Step 3>

[0988] Synthesis of 4-(3-((3-((tert-Butoxycarbonyl)amino)propyl)amino)-3-oxopropyl)benzyl 4-methylbenzenesulfonate (EX8-IM-3) Dissolve the compound (400 mg) obtained in <Step 2> of (Example 8) in tetrahydrofuran (8.0 mL). Add p-toluenesulfonyl chloride (272 mg), N,N-dimethyl-4-aminopyridine (15 mg), and triethylamine (0.33 mL), and stir at 70 °C for 6 hours. Add ethyl acetate (25 mL) and water (10 mL) to the reaction solution, separate the layers, and extract the aqueous layer with ethyl acetate (5 mL). Combine the organic layers, wash successively with semi-saturated sodium bicarbonate solution (10 mL), water (10 mL), and saturated brine (5 mL), dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Triturate the residue with tert-butyl methyl ether / heptane, filter the resulting solid, and wash with heptane to obtain the title compound (224 mg) as a light beige solid.

[0989] <Step 4>

[0990] Synthesis of S-(4-(3-((3-((tert-Butoxycarbonyl)amino)propyl)amino)-3-oxopropyl)benzyl)thioacetate (EX8-IM-4) Suspend the compound (224 mg) obtained in <Step 3> of (Example 8) in acetonitrile (4.5 mL). Add potassium thioacetate (87 mg), and stir at room temperature for 30 minutes. Add ethyl acetate (20 mL) and water (10 mL) to the reaction solution, separate the layers. Wash the organic layer successively with water (10 mL) and saturated brine (5 mL), dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography (10% ethyl acetate / heptane to ethyl acetate) to obtain the title compound (189 mg) as a white solid.

[0991] <Step 5>

[0992] Synthesis of S-(4-(3-((3-aminopropyl)amino)-3-oxopropyl)benzyl)thioacetate hydrochloride (EX8-IM-5) Using the compound (189 mg) obtained in <Step 4> of (Example 8), perform the same operations as in <Step 3> of (Example 7) to obtain the title compound (140 mg) as a white solid.

[0993] <Step 6>

[0994] Synthesis of alginic acid (AL-EX-8) incorporating S-(4-(3-((3-aminopropyl)amino)-3-oxopropyl)benzyl)thioacetate group was carried out using an aqueous solution (20 mL) of sodium alginate (manufactured by Kimica Corporation, ALG-2) prepared to be 1 wt% and the compound (67 mg) obtained in <Step 5> of (Example 8), and performing the same operations as in <Step 3> of (Example 1) to obtain the title compound (189 mg) in the form of a white solid.

[0995] (Example 9)

[0996] Synthesis of alginic acid (AL-EX-9) incorporating S-(2-(4-aminobutanamido)ethyl)thiobenzoate group

[0997] [Chemical Formula 116]

[0998]

[0999] <Step 1>

[1000] Synthesis of S-(2-((tert-butoxycarbonyl)amino)ethyl)thiobenzoate (EX9-IM-1)

[1001] Suspend 2-aminoethane-1-thiol hydrochloride (3.0 g) and triethylamine (4.1 mL) in dichloromethane (20 mL). Add a solution of di-tert-butyl dicarbonate (6.3 g) in dichloromethane (10 mL) under ice-water cooling and stir at room temperature for 2 hours. Add triethylamine (4.4 mL) and benzoyl chloride (3.7 mL) under ice-water cooling and stir at room temperature for 1 hour. Add tert-butyl methyl ether (100 mL) and water (50 mL) to the reaction solution, separate the layers, and extract the aqueous layer with tert-butyl methyl ether (50 mL). Combine the organic layers, wash successively with water (50 mL) and saturated brine (20 mL), dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Purify the residue by silica gel column chromatography (heptane to 30% ethyl acetate / heptane) to obtain the title compound (5.2 g) in the form of a colorless oil.

[1002] <Step 2>

[1003] Synthesis of S-(2-aminoethyl)thiobenzoate hydrochloride (EX9-IM-2)

[1004] To the compound (1.0 g) obtained in <Step 1> of (Example 9), 4 N hydrochloric acid / 1,4-dioxane (5.0 mL) was added, and the mixture was stirred at room temperature for 50 minutes. 4 N hydrochloric acid / 1,4-dioxane (1.0 mL) was added, and the mixture was stirred at room temperature for 1 hour. Further, 4 N hydrochloric acid / 1,4-dioxane (1.0 mL) was added, and the mixture was stirred at room temperature for 1.5 hours. Diisopropyl ether (14.0 mL) was added to the reaction solution. The precipitated solid was collected by filtration, washed with diisopropyl ether, and dried under reduced pressure to obtain the title compound (0.76 g) as a white solid.

[1005] <Step 3>

[1006] Synthesis of S-(2-(4-((tert-butoxycarbonyl)amino)butanamido)ethyl) thiobenzoate (EX9-IM-3) 4-((tert-Butoxycarbonyl)amino)butanoic acid (0.50 g) and triethylamine (0.36 mL) were dissolved in tetrahydrofuran (10.0 mL). Isobutyl chloroformate (0.34 mL) was added under ice-water cooling, and the mixture was stirred at the same temperature for 20 minutes. The compound (0.64 g) obtained in <Step 2> of (Example 9) and triethylamine (0.75 mL) were added at the same temperature, and the mixture was stirred at the same temperature for 1.5 hours. Ethyl acetate (20 mL) and water (10 mL) were added to the reaction solution, and the layers were separated. The organic layer was washed successively with semi-saturated aqueous sodium hydrogen carbonate (10 mL), water (10 mL), and saturated brine (5 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (heptane to 80% ethyl acetate / heptane) to obtain the title compound (0.74 g) as a white solid.

[1007] <Step 4>

[1008] Synthesis of S-(2-(4-aminobutanamido)ethyl) thiobenzoate hydrochloride (EX9-IM-4) Using the compound (0.74 g) obtained in <Step 3> of (Example 9), the same operation as in <Step 3> of (Example 7) was carried out to obtain the title compound (0.59 g) as a white solid.

[1009] <Step 5>

[1010] Synthesis of alginic acid incorporating S-(2-(4-aminobutanamido)ethyl) thiobenzoate group (AL-EX-9) Using an aqueous solution (20 mL) of sodium alginate (manufactured by Kimika Co., Ltd., ALG-2) prepared to be 1 wt% and the compound (61 mg) obtained in <Step 4> of (Example 9), the same operation as in <Step 3> of (Example 1) was carried out to obtain the title compound (198 mg) as a white solid.

[1011] (Example 9.1)

[1012] Preparation of alginic acid (EX-ALG-9.1) into which 4-((2-mercaptoethyl)amino)-4-oxobutylamino is introduced

[1013] [Chemical Formula 117]

[1014]

[1015] Dissolve the compound (25 mg) obtained in <Step 5> of (Example 9) in water (2.48 mL), add 1 N - aqueous sodium hydroxide solution (17 μL), and stir at 25 °C for 2 hours to prepare a 1 wt% solution of the title compound.

[1016] (Example 10)

[1017] Synthesis of alginic acid (AL-EX-10) into which S-benzoyl-N-glycyl-L-cysteine methyl ester group is introduced

[1018] [Chemical Formula 118]

[1019]

[1020] <Step 1>

[1021] Synthesis of (tert-butoxycarbonyl)-L-cysteine methyl ester (EX-10-IM-1)

[1022] According to a method known from the literature (Chem. Commun. (2012) 48: 7310 - 7312), to a mixture of commercially available L-cysteine methyl ester hydrochloride [CAS: 18598 - 63 - 5] (1 g) and tetrahydrofuran (7.5 mL), 2.3 M - aqueous sodium bicarbonate solution (7.5 mL) was added under ice-cooling and stirring. Then, di-tert-butyl dicarbonate (1.29 mL) was added at the same temperature, and the mixture was stirred at room temperature for 70 hours. After completion of the reaction, the solvent was distilled off under reduced pressure, and the residue was acidified with 1 N - hydrochloric acid (10 mL). The solution was extracted twice with ethyl acetate (20 mL), and the organic layer was washed successively with water (10 mL) and saturated brine (10 mL), and dried over anhydrous sodium sulfate. After filtering the organic layer, it was concentrated under reduced pressure to obtain a crude product of the title compound (1.308 g).

[1023] <Step 2>

[1024] Synthesis of S-benzoyl-N-(tert-butoxycarbonyl)-L-cysteine methyl ester (EX-10-IM-2)

[1025] According to the method known in the references (Synthesis (2017) 49: 4879-4886), to a mixture of p-benzoyl chloride (0.39 mL) and dichloromethane (4 mL), a mixture of the compound EX10-IM-1 (0.4 g) obtained in <Step 1> of Example 10, triethylamine (0.95 mL) and dichloromethane (4 mL) was added at room temperature, and the mixture was stirred at the same temperature for 1 hour and 30 minutes. After the reaction was completed, water (5 mL) and 1 N hydrochloric acid (5 mL) were added, and the mixture was extracted 3 times with ethyl acetate (20 mL) and washed with saturated brine (5 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (5% ethyl acetate / heptane to 40% ethyl acetate / heptane) to obtain the title compound (0.499 g) as a colorless oil.

[1026] <Step 3>

[1027] Synthesis of S-benzoyl-L-cysteine methyl ester hydrochloride (EX-10-IM-3)

[1028] To a mixture of the compound EX10-IM-2 (0.499 g) obtained in <Step 2> of Example 10 and 1,4-dioxane (3.49 mL), 4 N hydrogen chloride / 1,4-dioxane (3.49 mL) was added with stirring under water cooling, and the mixture was stirred at room temperature for 8 hours. After the reaction was completed, diisopropyl ether (40 mL) was added, and the precipitate was filtered. The recovered solid was dried under reduced pressure to obtain the title compound (0.376 g) as a white solid.

[1029] <Step 4>

[1030] Synthesis of S-benzoyl-N-((tert-butoxycarbonyl)glycyl)-L-cysteine methyl ester (EX-10-IM-4) To a mixture of the compound EX10-IM-3 (100 mg) obtained in <Step 3> of Example 10 and tetrahydrofuran (2000 μL), commercially available 2,5-dioxopyrrolidin-1-yl (tert-butoxycarbonyl)glycinate [CAS: 3392-07-2] (98.7 mg) and 1 M aqueous sodium bicarbonate (362.6 μL) were added with stirring under ice cooling, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was extracted 3 times with ethyl acetate (10 mL) and washed successively with water (5 mL) and saturated brine (5 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (25% ethyl acetate / heptane to 100% ethyl acetate, ethyl acetate to 20% methanol / ethyl acetate) to obtain a fraction containing the title compound (124 mg).

[1031] <Step 5>

[1032] Synthesis of S-benzoyl-N-glycyl-L-cysteine methyl ester hydrochloride (EX-10-IM-5)

[1033] Using the component containing compound EX10-IM-4 (0.11 g) obtained in <Step 4> of (Example 10), the same deprotection operation as <Step 4> of (Example 9) was carried out, and thus the title compound (0.08 g) was obtained in the form of a white solid.

[1034] <Step 6>

[1035] Synthesis of alginic acid (AL-EX-10) incorporating S-benzoyl-N-glycyl-L-cysteine methyl ester group: To an aqueous solution of sodium alginate (manufactured by Morita Pharmaceutical Co., Ltd., A-2) (19.78 mL) prepared to be 1 wt%, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (45.76 mg) and 1 M aqueous sodium bicarbonate solution (45.8 μL) were added at room temperature. Subsequently, a mixture of the compound EX10-IM-5 (15.23 mg) obtained in <Step 5> of (Example 10), water (1 mL) and ethanol (1 mL) was added dropwise at the same temperature. The reaction mixture was stirred at 40 °C for 4 hours. After adding sodium chloride (200 mg), ethanol (39.6 mL) was added, and the mixture was stirred at room temperature for 30 minutes. The resulting precipitate was collected by filtration, washed with ethanol, and dried under reduced pressure to obtain the title compound (181 mg) in the form of a white solid.

[1036] (Example 10.1)

[1037] Preparation of alginic acid (AL-EX-10.1) incorporating glycyl-L-cysteine methyl ester group

[1038] [Chemical formula 119]

[1039]

[1040] The compound (50 mg) obtained in <Step 6> of (Example 10) was dissolved in water (2.48 mL), 1 N aqueous sodium hydroxide solution (21.2 μL) was added, and the mixture was stirred at room temperature for 2 hours to prepare a 2 wt% solution of the title compound.

[1041] (Example 11)

[1042] Synthesis of alginic acid (AL-EX-11) incorporating N-(L-phenylamino)-S-benzoyl-L-cysteine group

[1043] [Chemical formula 120]

[1044]

[1045] <Step 1>

[1046] Synthesis of N-((tert-butoxycarbonyl)-L-phenylalanyl)-S-trityl-L-cysteine (EX11-IM-1) To a mixture of (tert-butoxycarbonyl)-L-phenylalanine [CAS: 13734-34-4] (200 mg), tetrahydrofuran (4 mL), and N-methylmorpholine (82.9 μL), isobutyl chloroformate (99 μL) was slowly added with stirring under ice cooling. The mixture was stirred at the same temperature for 30 minutes. Then, commercially available S-trityl-L-cysteine [CAS: 2799-07-7] (274 mg) was added with stirring under ice cooling, and the mixture was stirred at the same temperature for 30 minutes and then at room temperature for 30 minutes. Thereafter, water (5 mL) and 1 N hydrochloric acid (5 mL) were added to stop the reaction. The reaction solution was extracted three times with ethyl acetate (10 mL), and the organic layer was washed successively with water (10 mL) and saturated brine (10 mL), and dried over anhydrous sodium sulfate. After filtering the organic layer, it was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (25% ethyl acetate / heptane to 100% ethyl acetate, ethyl acetate to 15% methanol / ethyl acetate) to obtain the title compound (397 mg) in the form of a white amorphous solid.

[1047] <Step 2>

[1048] Synthesis of N-(L-phenylalanyl)-S-benzoyl-L-cysteine trifluoroacetate (EX11-IM-2) To a mixture of the compound EX11-IM-1 (100 mg) obtained in <Step 1> of (Example 11) and dichloromethane (500 μL), trifluoroacetic acid (1500 μL) and triisopropylsilane (35.4 μL) were added with stirring under ice cooling, and the mixture was stirred at room temperature for 30 minutes. To this mixture, benzoyl chloride (38 μL) was added dropwise with stirring under ice cooling. After stirring at room temperature for 25 hours, benzoyl chloride (19 μL) was added and stirred at the same temperature for a further 5 hours. After completion of the reaction, diisopropyl ether (20 mL) was added, and the precipitate was filtered. Since the filtration was incomplete, the filtrate was concentrated, diisopropyl ether (20 mL) was added, and the mixture was stirred at room temperature for 30 minutes. The precipitate was filtered and dried under reduced pressure to obtain the title compound (27.5 mg) in the form of a pale yellow solid.

[1049] <Step 3>

[1050] Using an aqueous solution (19.78 mL) of sodium alginate (manufactured by Morita Pharmaceutical Co., Ltd., A-2) prepared at 1% by weight and the compound EX10-IM-5 (15.23 mg) obtained in <Step 2> of (Example 11), the same operation as <Step 6> of (Example 10) was carried out, whereby the title compound (179 mg) was obtained in the form of a white solid.

[1051] (Example 11.1)

[1052] Preparation of alginic acid (AL-EX-11.1) into which L-phenylalanyl-L-cysteine sodium group is introduced

[1053] [Chemical Formula 121]

[1054]

[1055] Dissolve the compound AL-EX-11 (50 mg) obtained in <Step 3> of (Example 11) in water (2.48 mL), add 1 N sodium hydroxide aqueous solution (18.2 μL), and stir at room temperature for 2 hours to prepare a 2% by weight solution of the title compound.

[1056] (Example 12)

[1057] Synthesis of alginic acid (AL-EX-12) into which S-benzoyl-N-glycylglycyl-L-cysteine group is introduced

[1058] [Chemical Formula 122]

[1059]

[1060] <Step 1>

[1061] Synthesis of N-(tert-Butoxycarbonyl)glycylglycyl-S-trityl-L-cysteine (EX12-IM-1) Relative to a mixture of commercially available (tert-butoxycarbonyl)glycylglycine [CAS: 31972-52-8] (0.3 g) and tetrahydrofuran (6 mL), under ice-cooling and stirring, triethylamine (0.2 mL) and isobutyl chloroformate (0.19 mL) were added, and the mixture was stirred at room temperature for 30 minutes. Relative to this mixture, under ice-cooling and stirring, commercially available S-trityl-L-cysteine [CAS: 2799-07-7] (0.52 g) and triethylamine (0.4 mL) were added, and the mixture was stirred at room temperature for 20 hours. Water (5 mL) and 1 N hydrochloric acid (5 mL) were added to stop the reaction, and the mixture was extracted 3 times with ethyl acetate (10 mL). The organic layer was washed successively with water (5 mL) and saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (25% ethyl acetate / heptane to 100% ethyl acetate, ethyl acetate to 20% methanol / ethyl acetate) to obtain the title compound (0.581 g) in the form of a white amorphous solid.

[1062] <Step 2>

[1063] Synthesis of S-benzoyl-N-glycylglycyl-L-cysteine trifluoroacetate (EX12-IM-2) The compound EX12-IM-1 (200 mg) obtained in <Step 1> of (Example 12) was dissolved in trifluoroacetic acid (4000 μL), and triisopropylsilane (74.8 μL) was added at room temperature. After the reaction mixture was stirred at room temperature for 10 minutes, benzoyl chloride (80.4 μL) was added at the same temperature. After stirring at room temperature for 17 hours, benzoyl chloride (80.4 μL) was added at the same temperature and stirring was continued for 4 hours. After completion of the reaction, diisopropyl ether (20 mL) was added under ice-cooling and stirring, and the mixture was stirred at room temperature for 3 days. The suspension was filtered, and the recovered solid was dried under reduced pressure to obtain the title compound (140 mg) in the form of a pale yellow solid.

[1064] <Step 3>

[1065] Synthesis of alginic acid (AL-EX-12) incorporating S-benzoyl-N-glycylglycyl-L-cysteine group: To an aqueous solution (49.44 mL) of sodium alginate (manufactured by Morita Pharmaceutical Co., Ltd., A-2) at 1 wt%, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (114.4 mg) and 1 M aqueous sodium hydrogen carbonate solution (114.4 μL) were added at room temperature. Subsequently, a mixture of the compound EX12-IM-2 (51.86 mg) obtained in <Step 2> of (Example 12), water (1 mL) and ethanol (1 mL) was added dropwise at the same temperature. The reaction mixture was stirred at room temperature for 17 hours. After adding sodium chloride (500 mg), ethanol (98.9 mL) was added and the mixture was stirred at room temperature for 30 minutes. The resulting precipitate was collected by filtration, washed with ethanol, and dried under reduced pressure to obtain the title compound (480 mg) as a white solid.

[1066] (Example 12.1)

[1067] Preparation of alginic acid (AL-EX-12.1) incorporating glycylglycyl-L-cysteine sodium group

[1068] [Chemical formula 123]

[1069]

[1070] The compound AL-EX-12 (50 mg) obtained in <Step 3> of (Example 12) was dissolved in water (2.48 mL), 1 N aqueous sodium hydroxide solution (19.4 μL) was added, and the mixture was stirred at room temperature for 2 hours to prepare a 2 wt% solution of the title compound.

[1071] (Example 13)

[1072] Synthesis of alginic acid (AL-EX-13) incorporating S-benzoyl-N-glycyl-L-cysteine group

[1073] [Chemical formula 124]

[1074]

[1075] <Step 1>

[1076] Synthesis of N-((tert-butoxycarbonyl)glycyl)-S-trityl-L-cysteine (EX13-IM-1) Relative to a mixture of commercially available S-trityl-L-cysteine [CAS: 2799-07-7] (0.27 g) and water (1 mL), at room temperature, a solution of commercially available 2,5-dioxopyrrolidin-1-yl (tert-butoxycarbonyl)glycinate [CAS: 3392-07-2] (0.2 g) in tetrahydrofuran (2 mL) was added, and the mixture was stirred at the same temperature for 1 hour and 30 minutes. Subsequently, 2,5-dioxopyrrolidin-1-yl (tert-butoxycarbonyl)glycinate (0.04 g) was added, and after further stirring for 30 minutes, ethyl acetate (20 mL) and 1 N hydrochloric acid (5 mL) were added for separation. The organic layer was washed successively with water (5 mL) and saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product (0.43 g) of the title compound in the form of a white amorphous solid.

[1077] <Step 2>

[1078] Synthesis of S-benzoyl-N-glycyl-L-cysteine trifluoroacetate (EX13-IM-2)

[1079] Relative to a mixture of the compound EX13-IM-1 (150 mg) obtained in <Step 1> of (Example 13) and dichloromethane (750 μL), trifluoroacetic acid (2250 μL) and triisopropylsilane (62.2 μL) were added with stirring under ice cooling. The mixture was stirred at room temperature for 30 minutes, and benzoyl chloride (66.9 μL) was added dropwise with stirring under ice cooling, followed by stirring at room temperature for 20 hours and 30 minutes. Subsequently, benzoyl chloride (133.8 μL) was added at room temperature, and after stirring at the same temperature for 24 hours, dichloromethane was concentrated under reduced pressure. Diisopropyl ether (20 mL) was added to the residue, and the suspension was stirred at room temperature overnight, filtered, and dried under reduced pressure to obtain the title compound (47.7 mg) in the form of a pale yellow solid.

[1080] <Step 3>

[1081] Synthesis of alginic acid (AL-EX-13) incorporating S-benzoyl-N-glycyl-L-cysteine Using an aqueous solution (49.44 mL) of sodium alginate (manufactured by Morita Pharmaceutical Co., Ltd., A-2) prepared to 1 wt% and the compound EX13-IM-5 (45.33 mg) obtained in <Step 2> of (Example 13), the same operation as <Step 3> of (Example 12) was carried out to obtain the title compound (468 mg) in the form of a white solid.

[1082] (Example 13.1)

[1083] Preparation of alginic acid (AL-EX-13.1) incorporating sodium glycyl-L-cysteinate

[1084] [Chemical formula 125]

[1085]

[1086] Dissolve the compound AL-EX-13 (50 mg) obtained in <Step 3> of (Example 13) in water (2.478 mL), add 1 N sodium hydroxide aqueous solution (21.8 μL), and stir at room temperature for 2 hours to prepare a 2 wt% solution of the title compound.

[1087] (Example 14)

[1088] Synthesis of alginic acid (AL-EX-14) incorporating N-(L-alanyl)-S-benzoyl-L-cysteinate

[1089] [Chemical formula 126]

[1090]

[1091] <Step 1>

[1092] Synthesis of N-((tert-butoxycarbonyl)L-alanyl)-S-trityl-L-cysteine (EX14-IM-1) Dissolve (tert-butoxycarbonyl)-L-alanine [CAS: 15761-38-3] (260.5 mg) in tetrahydrofuran (20 mL). To this solution, slowly add N-methylmorpholine (151 μL) and isobutyl chloroformate (181 μL) with stirring under ice-cooling. Stir the mixture at the same temperature for 25 minutes. Then, with stirring under ice-cooling, add commercially available S-trityl-L-cysteine [CAS: 2799-07-7] (500.3 mg), stir at the same temperature for 1 hour, and then at room temperature for 1 hour and 40 minutes. After that, add water (10 mL) and 1 N hydrochloric acid (10 mL) to stop the reaction. Extract the reaction solution with ethyl acetate (10 mL) three times, wash the organic layer successively with water (5 mL) and saturated brine (5 mL), and dry over anhydrous sodium sulfate. After filtering the organic layer, concentrate it under reduced pressure, and purify the resulting crude product by silica gel column chromatography (25% ethyl acetate / heptane to 100% ethyl acetate, ethyl acetate to 15% methanol / ethyl acetate) to obtain the title compound (0.674 g) as a white amorphous solid.

[1093] <Step 2>

[1094] Synthesis of N-(L-alanyl)-S-benzoyl-L-cysteine trifluoroacetate (EX14-IM-2) The compound EX14-IM-1 (0.674 g) obtained in <Step 1> of (Example 14) was dissolved in trifluoroacetic acid (6.1 mL) under ice-cooling. The reaction mixture was stirred at room temperature for 1 hour and 30 minutes. Triisopropylsilane (0.26 mL) was added under ice-cooling, and the mixture was stirred at room temperature for 30 minutes. Then, benzoyl chloride (0.335 mL) was added dropwise with stirring under ice-cooling, and the mixture was stirred at room temperature for 17 hours. Further, benzoyl chloride (0.335 mL) was added with stirring under ice-cooling, and the mixture was stirred at room temperature for 3 hours. Diisopropyl ether (80 mL) was added, and the suspension was stirred at room temperature overnight. After filtration, it was dried under reduced pressure to obtain the title compound (0.271 g) as a beige solid.

[1095] <Step 3>

[1096] Synthesis of alginic acid (AL-EX-14) incorporating N-(L-alanyl)-S-benzoyl-L-cysteine group To an aqueous solution (39.5 mL) of sodium alginate (manufactured by Morita Pharmaceutical Co., Ltd., A-2) prepared to be 1 wt%, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (121.8 mg) and 1 M aqueous sodium bicarbonate solution (91 μL) were added with stirring under water-cooling. Then, a mixture of the compound EX14-IM-2 (37.5 mg) obtained in <Step 2> of (Example 14), water (1 mL), and ethanol (1 mL) was added dropwise at the same temperature. The reaction mixture was stirred at room temperature for 66 hours. After adding sodium chloride (400 mg), ethanol (79 mL) was added, and the mixture was stirred at room temperature for 30 minutes. The resulting precipitate was collected by filtration, washed with ethanol, and dried under reduced pressure. The obtained solid was dissolved in water, filtered, and then freeze-dried to obtain the title compound (0.325 g) as a white solid.

[1097] (Example 14.1)

[1098] Preparation of alginic acid (AL-EX-14.1) incorporating sodium L-alanyl-L-cysteine group

[1099] [Chemical formula 127]

[1100]

[1101] The compound AL-EX-14 (100 mg) obtained in <Step 3> of (Example 14) was dissolved in water (4957 μL), 1 N aqueous sodium hydroxide solution (42 μL) was added, and the mixture was stirred at room temperature for 2 hours to prepare a 2 wt% solution of the title compound.

[1102] (Example 15)

[1103] Synthesis of alginic acid (AL-EX-15) incorporating N-(3-aminopropionyl)-S-benzoyl-L-cysteine

[1104] [Chemical Formula 128]

[1105]

[1106] <Step 1>

[1107] Synthesis of N-(3-((tert-butoxycarbonyl)amino)propionyl)-S-trityl-L-cysteine (EX15-IM-1) Using commercially available N-(tert-butoxycarbonyl)-β-alanine [CAS: 3303-84-2] (260.5 mg), tetrahydrofuran (20 mL), N-methylmorpholine (151 μL), isobutyl chloroformate (181 μL), and commercially available S-trityl-L-cysteine [CAS: 2799-07-7] (500 mg), the same operation as <Step 1> of (Example 14) was carried out, and thus the title compound (0.445 g) was obtained in the form of a white amorphous solid.

[1108] <Step 2>

[1109] Synthesis of N-(3-aminopropionyl)-S-benzoyl-L-cysteine trifluoroacetate (EX15-IM-2) The compound EX15-IM-1 (0.445 g) obtained in <Step 1> of (Example 15) was dissolved in trifluoroacetic acid (8.9 mL) under ice cooling. The reaction mixture was stirred at room temperature for 70 minutes. Triisopropylsilane (0.18 mL) was added under ice cooling, and the mixture was stirred at room temperature for 20 minutes. Then, benzoyl chloride (0.19 mL) was added at room temperature, and the mixture was stirred at room temperature for 17 hours and 50 minutes. Further, benzoyl chloride (0.095 mL) was added at room temperature, and after stirring at the same temperature for 4 hours, an equal amount of benzoyl chloride was added, and the mixture was stirred at room temperature for 18 hours. Then, benzoyl chloride (0.19 mL) was added at room temperature, and the mixture was stirred at the same temperature for 2 hours and 40 minutes. Diisopropyl ether (50 mL) was added, and the suspension was stirred at room temperature for 4 hours, filtered, and dried under reduced pressure, thereby obtaining a mixture of EX15-IM-1 and the title compound (0.445 g). A part of this mixture (0.1876 g) was dissolved in trifluoroacetic acid (1.69 mL). Then, benzoyl chloride (0.18 mL) was added at room temperature, and the mixture was stirred at the same temperature for 3 hours. Diisopropyl ether (20 mL) was added, and the suspension was stirred at room temperature overnight, filtered, and dried under reduced pressure, thereby obtaining the title compound (0.1055 g) in the form of a white solid.

[1110] <Step 3>

[1111] Synthesis of alginic acid (AL-EX-15) incorporating N-(3-aminopropionyl)-S-benzoyl-L-cysteine Using an aqueous solution (39.5 mL) of sodium alginate (manufactured by Mochida Pharmaceutical Co., Ltd., A-2) prepared to 1% by weight and the compound EX15-IM-2 (37.5 mg) obtained in <Step 3> of (Example 15), the same operations as in <Step 3> of (Example 14) were carried out, whereby the title compound (0.348 g) was obtained as a white solid.

[1112] (Example 15.1)

[1113] Preparation of alginic acid (AL-EX-15.1) incorporating (3-aminopropionyl)-L-cysteine sodium

[1114] [Chemical formula 129]

[1115]

[1116] The compound AL-EX-15 (100 mg) obtained in <Step 3> of (Example 15) was dissolved in water (4957 μL), 1 N sodium hydroxide aqueous solution (42 μL) was added, and the mixture was stirred at room temperature for 2 hours to prepare a 2% by weight solution of the title compound.

[1117] (Example 16)

[1118] Synthesis of alginic acid (AL-EX-16) incorporating S-benzoyl-N-glycyl-L-prolyl-L-cysteine

[1119] [Chemical formula 130]

[1120]

[1121] <Step 1>

[1122] Synthesis of N-(tert-butoxycarbonyl)glycyl-L-prolyl-S-trityl-L-cysteine (EX16-IM-1) Using commercially available (tert-butoxycarbonyl)glycyl-L-proline [CAS: 14296-92-5] (374.3 mg), tetrahydrofuran (20 mL), N-methylmorpholine (151 μL), isobutyl chloroformate (181 μL), and commercially available S-trityl-L-cysteine [CAS: 2799-07-7] (500 mg), the same operations as in <Step 1> of (Example 14) were carried out, whereby the title compound (0.453 g) was obtained as a white amorphous solid.

[1123] <Step 2>

[1124] S-Benzoyl-N-glycyl-L-prolyl-L-cysteine trifluoroacetate (EX16-IM-2) was synthesized using the compound EX16-IM-1 (0.453 g) obtained in <Step 1> of (Example 16), trifluoroacetic acid (4.1 mL), triisopropylsilane (0.15 mL), and benzoyl chloride (0.45 mL), and performing the same operations as in <Step 2> of (Example 14), whereby the title compound (0.2251 g) was obtained as a beige solid.

[1125] <Step 3>

[1126] Synthesis of alginic acid (AL-EX-16) incorporating S-benzoyl-N-glycyl-L-prolyl-L-cysteine group Using an aqueous solution (39.5 mL) of sodium alginate (manufactured by Morita Pharmaceutical Co., Ltd., A-2) prepared to be 1 wt% and the compound EX16-IM-2 (45.1 mg) obtained in <Step 3> of (Example 16), and performing the same operations as in <Step 3> of (Example 14), whereby the title compound (0.376 g) was obtained as a white solid.

[1127] (Example 16.1)

[1128] Preparation of alginic acid (AL-EX-16.1) incorporating sodium glycyl-L-prolyl-L-cysteine group

[1129] [Chemical formula 131]

[1130]

[1131] Dissolve the compound AL-EX-16 (30 mg) obtained in <Step 3> of (Example 16) in water (1484 μL), add 1 N sodium hydroxide aqueous solution (16 μL), and stir at room temperature for 2 hours to prepare a 2 wt% solution of the title compound.

[1132] [Table 2]

[1133] Physical property data of alginic acid derivatives

[1134] [Table 3-1]

[1135] NMR data of intermediate compounds

[1136] [Table 3-2]

[1137]

[1138] [Table 3-3]

[1139]

[1140] [Table 4]

[1141] LC-MS data of intermediate compounds

[1142] <Determination of the introduction rate of reactive groups>

[1143] The introduction rate of reactive groups is the value expressed as a percentage of the number of reactive groups introduced per monosaccharide unit of guluronic acid, which is a repeating unit of alginic acid.

[1144] In the examples of this specification, the introduction rate (mol%) of reactive groups or complementary reactive groups is calculated by 1 the integral ratio of 1H-NMR. Moreover, the amount of alginic acid necessary for calculating the introduction rate is determined by the carbazole-sulfuric acid method using a calibration curve, and the amount of reactive groups or complementary reactive groups can also be determined by the absorbance measurement method using a calibration curve.

[1145] <Determination of molecular weight>

[1146] Weigh the alginic acid solid with introduced crosslinking groups obtained in the examples, add ultrapure water, and prepare a 1% aqueous solution. Then, dilute it so that the alginic acid concentration reaches 0.2%, and the solution composition is a 10 mmol / L phosphate buffer (pH 7.4) containing 0.15 mol / L of NaCl. After removing insolubles from this solution by passing it through a Minisart HighFlow filter (manufactured by SARTORIUS) made of polyethersulfone with a pore size of 0.45 μm or 0.22 μm, supply 200 μL of this solution to a Superose6 Increase 10 / 300GL column (manufactured by GE Healthcare Sciences) and perform gel filtration. Gel filtration is carried out using an AKTA Explorer 10S as a chromatographic device, using a 10 mmol / L phosphate buffer (pH 7.4) containing 0.15 mol / L of NaCl as an eluent, and under the condition of a flow rate of 0.8 mL / min at room temperature. Detect the absorption at 220 nm, 240 nm, or 235 nm and prepare a chromatogram for each sample. In addition, as another method, detect the absorption at 215 nm. The peak analysis of the obtained chromatogram is carried out using Unicorn 5.31 software (manufactured by GE Healthcare Sciences).

[1147] The molecular weight of the alginic acid into which a crosslinking group is introduced is determined using blue dextran (molecular weight: 2 million Da, manufactured by SIGMA), thyroglobulin (molecular weight: 669,000 Da, manufactured by GE Healthcare), ferritin (molecular weight: 440,000 Da, manufactured by GE Healthcare), aldolase (molecular weight: 158,000 Da, manufactured by GE Healthcare), conalbumin (molecular weight: 75,000 Da, manufactured by GE Healthcare), ovalbumin (molecular weight: 44,000 Da, manufactured by GE Healthcare), ribonuclease A (molecular weight: 13,700 Da, manufactured by GE Healthcare), and aprotinin (molecular weight: 6,500 Da, manufactured by GE Healthcare) as standards. A calibration curve is prepared from the liquid volume and molecular weight of each component at the absorption peak at 280 nm during gel filtration under the same conditions. Two calibration curves are prepared, one from blue dextran to ferritin and the other from ferritin to aprotinin. Using this calibration curve, the molecular weight (Mi) at the elution time i of the obtained chromatogram is first calculated. Next, the absorbance at the elution time i is read as Hi, and the weight-average molecular weight (Mw) is determined from these data using the following formula.

[1148] [Equation 1]

[1149]

[1150] The molecular weight of the alginic acid before introducing the reactive group is determined as follows. That is, considering the dry weight loss, each sodium alginate is weighed and ultrapure water is added to prepare a 1% aqueous solution. Next, it is diluted so that the alginic acid concentration reaches 0.2% and the solution composition is 10 mmol / L phosphate buffer (pH 7.4) containing 0.15 mol / L NaCl. After removing insolubles using a hydrophilic PVDF Mylex GV33 filter (manufactured by MERCK-Millipore) with a pore size of 0.22 μm, 200 μL is subjected to gel filtration and gel filtration is carried out under the same conditions as for the alginic acid into which the crosslinking group is introduced. Detection is carried out using a differential refractometer. In addition, as another method, insolubles are removed using a polyethersulfone Minisart High Flow filter (manufactured by SARTORIUS) with a pore size of 0.45 μm.

[1151] The weight-average molecular weight of the sodium alginate before introducing the crosslinking group is determined using the same method as the calculation method for the molecular weight of the alginic acid into which the crosslinking group is introduced. Among them, Hi is calculated from the data of the differential refractometer.

[1152] (Examples 1) to (Example 3), (Examples 7) to (Example 9) The molecular weight of the sodium alginate (manufactured by Kimika Co., Ltd., ALG-2) before introducing the crosslinking group eluted in a wide range from 2.6 million Da to 145,000 Da, and the weight-average molecular weight was calculated to be 1.46 million Da.

[1153] (Examples 4) to (Example 6) The molecular weight of sodium alginate (manufactured by Kimica Corporation, ALG-2) before introducing a crosslinking group was eluted in a wide range of 9,600 Da to 2.51 million Da, and the weight average molecular weight was calculated to be 1.38 million Da.

[1154] (Examples 10) to (Example 16.1) The molecular weight of sodium alginate (manufactured by Motoda Pharmaceutical Co., Ltd., A-2) before introducing a crosslinking group was the molecular weight described in Table 1 above.

[1155] <Measurement of gel stability (1)>

[1156] The alginic acid derivative (AL-EX-2) prepared by the same method as in <Step 3> of (Example 2) was dissolved in water at a concentration of 0.5% by weight to obtain an alginic acid aqueous solution (2-1).

[1157] Further, 3 volumes of phosphate buffered saline (PBS) were added to 2% by weight of the alginic acid derivative (AL-EX-7.1) solution, (AL-EX-10.1) solution, (AL-EX-11.1) solution, (AL-EX-12.1) solution, and (AL-EX-13.1) solution prepared by the same methods as in (Example 7), (Example 10), (Example 11), (Example 12), and (Example 13) to form 0.5% by weight, and alginic acid aqueous solutions (7-1), (10-1), (11-1), (12-1), and (13-1) were obtained.

[1158] The alginic acid aqueous solution (2-1) and the alginic acid aqueous solutions (7-1), (10-1), (11-1), (12-1), or (13-1) were mixed in equal amounts of 250 μL each, 40 mL of a 30 mmol / L calcium chloride solution was added, and the mixture was stirred for 5 minutes to obtain an alginic acid gel. The gel was washed once with 10 mL of PBS to obtain a chemically crosslinked alginic acid gel. 19.5 mL of PBS was added to the gel, and the mixture was shaken at 37°C. The aqueous solution was recovered over time, and an amount of PBS equal to the recovered amount was added. After the test was completed, 10 μL of alginic acid lyase (Nippon Gene, 319-08261) was added to the test solution, and the mixture was shaken overnight at 37°C to completely disintegrate the gel, and the aqueous solution was recovered. The alginic acid concentration in the recovered aqueous solution was measured by the carbazole sulfuric acid method, and the value obtained by dividing the value obtained by correcting the alginic acid amount in the aqueous solution at each time by the recovered alginic acid amount by the total alginic acid amount calculated from the alginic acid concentration at all times and the alginic acid concentration after the test was expressed as a percentage, and the obtained value was used as the disintegration rate and used as an index of gel stability.

[1159] Obtained Figure 1The results. The disintegration rate of the aforementioned crosslinked alginate gel (beads) after 24 hours was 1% or less, and the disintegration rate after 96 hours was 40% or less. From this, the stability of the gel could be confirmed. That is, it indicates that the (bead) structure prepared by forming a chemical crosslink based on the Michael reaction can maintain its structure over a long period of time.

[1160] <Determination of gel stability (2)>

[1161] The alginic acid aqueous solutions (2-1) and (7-1), (10-1), (11-1), (12-1) or (13-1) obtained in <Determination of gel stability (1)> were each mixed in an equal amount of 250 μL, 40 mL of a calcium chloride solution with a concentration of 30 mmol / L was added, and the mixture was stirred for 5 minutes to obtain an alginate gel. The gel was washed once with 10 mL of physiological saline to obtain a chemically crosslinked alginate gel. 19.5 mL of a 5 mM disodium ethylenediaminetetraacetate dihydrate (EDTA·2K) / physiological saline solution was added to the gel, and after shaking at 37°C for 24 hours, the aqueous solution was recovered, and a 5 mM EDTA·2K / physiological saline solution equal to the recovered amount was added. After the test was completed, 10 μL of alginate lyase (Nippon Gene, 319-08261) was added to the test solution, and the gel was completely disintegrated by shaking overnight at 37°C, and the aqueous solution was recovered. The alginic acid concentration in the recovered aqueous solution was measured by the carbazole-sulfuric acid method, and the value obtained by dividing the value obtained by correcting the amount of alginic acid in the aqueous solution at each time by the amount of alginic acid already recovered by the total amount of alginic acid calculated from the alginic acid concentration at all times and the alginic acid concentration after the test was completed was expressed as a percentage, and the obtained value was used as the disintegration rate and used as an index of gel stability.

[1162] obtained Figure 2 The results. The disintegration rate of the aforementioned crosslinked alginate gel (beads) was approximately 40% even after 24 hours, and the stability of the gel could be confirmed. That is, it indicates that the (bead) structure prepared even after EDTA treatment can maintain its structure over a long period of time.

[1163] <Determination of gel stability (3)>

[1164] The alginic acid derivative (AL-EX-5) prepared by the same method as <Step 3> of (Example 5) was dissolved in water at a concentration of 1.0% by weight to obtain an alginic acid aqueous solution (5-1). Further, PBS was added in equal amounts to the 2% by weight alginic acid derivative (AL-EX-7.1) solution prepared by the same method as (Example 7) and the (AL-EX-10.1) solution, (AL-EX-11.1) solution, (AL-EX-12.1) solution, and (AL-EX-13.1) solution obtained in (Example 10), (Example 11), (Example 12), and (Example 13) to form 1.0% by weight, obtaining alginic acid aqueous solutions (7-2), (10-2), (11-2), (12-2), and (13-2).

[1165] The alginic acid aqueous solution (5-1) and the alginic acid aqueous solutions (7-2), (10-2), (11-2), (12-2), or (13-2) were mixed in equal amounts of 250 μL each, 40 mL of a calcium chloride solution with a concentration of 30 mmol / L was added, and the mixture was stirred for 5 minutes to obtain an alginic acid gel. The gel was washed once with 10 mL of PBS to obtain a chemically crosslinked alginic acid gel. 19.5 mL of PBS was added to the gel, and the mixture was shaken at 37°C. The aqueous solution was recovered over time, and PBS equal to the recovered amount was supplemented. After the test was completed, 10 μL of alginic acid lyase (Nippon Gene, 319-08261) was added to the test solution, and the mixture was shaken overnight at 37°C to completely disintegrate the gel, and the aqueous solution was recovered. The alginic acid concentration in the recovered aqueous solution was measured by the carbazole-sulfuric acid method, and the value obtained by dividing the value obtained by correcting the alginic acid amount in the aqueous solution at each time by the recovered alginic acid amount by the total alginic acid amount calculated from the alginic acid concentration at all times and the alginic acid concentration after the test was completed was expressed as a percentage, and the obtained value was used as the disintegration rate, which was used as an index of gel stability.

[1166] The results are shown in Figure 3 . The disintegration rate of each of the aforementioned crosslinked alginic acid gels (beads) after 96 hours was 26% or less. It should be noted that the disintegration rate of the crosslinked alginic acid gel prepared using (AL-EX-7.1) and (AL-EX-2) as controls was 28.8% at the 96-hour time point, indicating stability equal to or higher than that of the control.

[1167] <Determination of Gel Stability (4)>

[1168] The aqueous alginic acid solutions (5-1) and (7-2), (10-2), (11-2), (12-2), or (13-2) obtained in <Determination of Gel Stability (3)> were each mixed in an equal amount of 250 μL, 40 mL of a calcium chloride solution with a concentration of 30 mmol / L was added, and the mixture was stirred for 5 minutes to obtain an alginic acid gel. The gel was washed once with 10 mL of physiological saline to obtain a chemically crosslinked alginic acid gel. 19.5 mL of a 5 mM EDTA·2K / physiological saline solution was added to the gel, and after shaking at 37 °C for 24 hours, the aqueous solution was recovered, and an amount of 5 mM EDTA·2K / physiological saline solution equal to the recovered amount was replenished. After the test, 10 μL of alginate lyase (Nippon Gene, 319-08261) was added to the test solution, and the gel was completely disintegrated by shaking overnight at 37 °C, and the aqueous solution was recovered. The alginic acid concentration in the recovered aqueous solution was measured by the carbazole-sulfuric acid method, and the value obtained by correcting the alginic acid amount in the aqueous solution at each time point with the amount of alginic acid already recovered was divided by the total alginic acid amount calculated from the alginic acid concentration at all time points and the alginic acid concentration after the end of the test, and the resulting value was expressed as a percentage. The obtained value was used as the disintegration rate and used as an index of gel stability.

[1169] obtained Figure 4 The results were as follows. The disintegration rate of the aforementioned crosslinked alginic acid gel (beads) was about 30% even after 24 hours, and the stability of the gel was confirmed. That is, it was shown that the (bead) structure prepared by EDTA treatment could maintain its structure even after a long time.

[1170] <Determination of Gel Stability (5)>

[1171] The alginic acid derivative (AL-EX-2) prepared by the same method as <Step 3> of (Example 2) was dissolved in water to a concentration of 1.0 wt% to obtain an aqueous alginic acid solution (2-3).

[1172] Furthermore, PBS was added in an equal amount to a 2 wt% alginic acid derivative (AL-EX-7.1) solution and a (AL-EX-16.1) solution prepared by the same method as (Example 7) to form 1.0 wt%, and aqueous alginic acid solutions (7-2) and (16-1) were obtained.

[1173] The aqueous alginate solution (2-3) and the aqueous alginate solution (7-2) or (16-1) were mixed in equal amounts of 300 μL each and pipetted 3 times. 500 μL of this solution was dispensed into 2.5 mL of a calcium chloride solution with a concentration of 55 mmol / L. After gently shaking, it was allowed to stand for 5 minutes. Further, 3.5 mL of a calcium chloride solution with the same concentration was added, and it was allowed to stand for 5 minutes. The gel was washed twice with 5 mL of physiological saline to obtain a chemically crosslinked alginate gel. 19.5 mL of PBS was added to this gel, and it was shaken at 37°C. The aqueous solution was recovered over time, and PBS equal to the amount recovered was replenished. After the test, 10 μL of alginate lyase (Nippon Gene, 319-08261) was added to the test solution, and it was shaken overnight at 37°C to completely disintegrate the gel, and the aqueous solution was recovered. The alginate concentration in the recovered aqueous solution was measured by the carbazole-sulfuric acid method. The value obtained by dividing the value obtained by correcting the alginate amount in the aqueous solution at each time by the amount of alginate already recovered by the total alginate amount calculated from the alginate concentration at all times and the alginate concentration after the end of the test was expressed as a percentage, and the obtained value was used as the disintegration rate, which was used as an index of gel stability.

[1174] obtained Figure 8 The results were as follows. The disintegration rate of the aforementioned crosslinked alginate gel (beads) after 24 hours was 15% or less, and the disintegration rate after 96 hours was 20% or less. From this, the stability of the gel was confirmed. That is, it was shown that the (bead) structure prepared by forming a chemical crosslink based on the Michael reaction could maintain its structure even after a long time.

[1175] <Measurement of Gel Stability (6)>

[1176] The alginate derivatives (AL-EX-2) and (AL-EX-5) prepared by the same method as in <Step 3> of (Example 2) and <Step 3> of (Example 5) were dissolved in water to a concentration of 1.0 wt% to obtain aqueous alginate solutions (2-1) and (5-1). Further, PBS was added in equal amounts to the 2 wt% alginate derivative (AL-EX-7.1) solution prepared by the same method as in (Example 7) and the (AL-EX-14.1) solution, (AL-EX-15.1) solution, and (AL-EX-16.1) solution obtained in (Example 14), (Example 15), and (Example 16) to form 1.0 wt%, and aqueous alginate solutions (7-2), (14-1), (15-1), and (16-1) were obtained.

[1177] The aqueous alginic acid solution (2-1) or (5-1) and the aqueous alginic acid solutions (7-2), (14-1), (15-1) or (16-1) were mixed in equal amounts of 300 μL respectively and pipetted several times. 500 μL of this solution was dispensed into a calcium chloride solution (2.5 mL) with a concentration of 55 mmol / L. After gently shaking, it was left standing for 5 minutes. Further, a calcium chloride solution (3.5 mL) with the same concentration was added and left standing for 5 minutes. The gel was washed twice with 5 mL of physiological saline to obtain a chemically crosslinked alginic acid gel. 19.5 mL of a 5 mM disodium ethylenediaminetetraacetate dihydrate (EDTA·2K) / physiological saline solution was added to this gel. After shaking at 37 °C for 24 hours, the aqueous solution was recovered, and a 5 mM EDTA·2K / physiological saline solution equal to the recovered amount was replenished. After the test, 10 μL of alginate lyase (Nippon Gene, 319-08261) was added to the test solution, and it was shaken overnight at 37 °C to completely disintegrate the gel, and the aqueous solution was recovered. The alginic acid concentration in the recovered aqueous solution was measured by the carbazole-sulfuric acid method. The value obtained by dividing the value obtained by correcting the alginic acid amount in the aqueous solution at each time by the amount of alginic acid already recovered by the total alginic acid amount calculated from the alginic acid concentration at all times and the alginic acid concentration after the end of the test was expressed as a percentage, and the obtained value was used as the disintegration rate and used as an index of gel stability.

[1178] obtained Figure 9 The results were as follows. The aforementioned crosslinked alginic acid gel (beads) had a disintegration rate of about 40% even after 24 hours, and the stability of the gel could be confirmed. That is, it was shown that the (bead) structure prepared by EDTA treatment could maintain its structure even after a long time.

[1179] <Measurement of Gel Transmittance>

[1180] The alginic acid derivatives (AL-EX-2) prepared by the same method as in <Step 3> of (Example 2) and the alginic acid derivatives (AL-EX-5) prepared by the same method as in <Step 3> of (Example 5) were dissolved in water to a concentration of 2.0% to prepare an aqueous alginic acid solution. 4 / 5 volume of fluorescein isothiocyanate-dextran with a molecular weight of 150,000 prepared at 1 mg / mL (Sigma-Aldrich, FD150S) and 2.2 volumes of PBS were added to this aqueous alginic acid solution to obtain 0.5% aqueous alginic acid solutions (2-2) and (5-2) containing 0.2 mg / mL fluorescein isothiocyanate-dextran.

[1181] Further, 3 volumes of PBS were added to 2% alginic acid derivative (AL-EX-7.1) solution, (AL-EX-10.1) solution, (AL-EX-11.1) solution, (AL-EX-12.1) solution, and (AL-EX-13.1) solution prepared by the same method as in (Example 7), (Example 10), (Example 11), (Example 12), and (Example 13) to form 0.5 wt%, obtaining aqueous alginic acid solutions (7-1), (10-1), (11-1), (12-1), and (13-1).

[1182] 250 μL of aqueous alginic acid solution (2-2) or (5-2) and aqueous alginic acid solutions (7-1), (10-1), (11-1), (12-1), or (13-1) were mixed in equal amounts, 40 mL of a 30 mmol / L calcium chloride solution was added, and the mixture was stirred for 5 minutes to obtain an alginic acid gel. The gel was washed once with 10 mL of physiological saline to obtain a chemically crosslinked alginic acid gel encapsulating fluorescein isothiocyanate-dextran. 19.5 mL of physiological saline was added to the gel, and the mixture was shaken at 37°C. The aqueous solution was recovered over time, and PBS equal to the recovered amount was replenished. After the test, 10 μL of alginate lyase (Nippon Gene, 319-08261) was added to the test solution, and the mixture was shaken at 37°C for 3 hours or more until the gel completely disintegrated, and the aqueous solution was recovered. The dextran concentration in the recovered aqueous solution was measured by fluorescence quantification (excitation light: 485 nm, fluorescence: 535 nm), and the value obtained by dividing the amount of dextran at each time by the amount of dextran after the end of the test was expressed as a percentage, and the obtained value was used as the transmittance.

[1183] The Figure 5 results were obtained. The transmittance after 3 hours was about 14 to about 20%. And the transmittance after 24 hours was about 30 to about 37%.

[1184] And the Figure 6 results were obtained. The transmittance after 3 hours was about 17 to about 27%. And the transmittance after 24 hours was about 31 to about 42%.

[1185] <Measurement of Gel Transmittance (2)>

[1186] An alginic acid aqueous solution was prepared by dissolving an alginic acid derivative (AL-EX-2) prepared by the same method as <Step 3> of (Example 2) in water at a concentration of 2.0% by weight. To this alginic acid aqueous solution (1.0 mL), fluorescein isothiocyanate-dextran with a molecular weight of 150,000 prepared at 1 mg / mL (Sigma-Aldrich, FD150S) (0.4 mL) and PBS (0.6 mL) were added to obtain a 1.0% alginic acid aqueous solution (2-4) containing 0.2 mg / mL fluorescein isothiocyanate-dextran.

[1187] Furthermore, PBS was added in equal amounts to 2% alginic acid derivative (AL-EX-7.1) solutions and (AL-EX-16.1) solutions prepared by the same methods as (Example 7) and (Example 16) to form 1.0% by weight, obtaining alginic acid aqueous solutions (7-2) and (16-1).

[1188] The alginic acid aqueous solution (2-4) and the alginic acid aqueous solution (7-2) or (16-1) were mixed in equal amounts of 300 μL each and pipetted 3 times. 500 μL of this solution was dispensed into a calcium chloride solution (2.5 mL) with a concentration of 55 mmol / L, gently shaken, and then allowed to stand for 5 minutes. Further, a calcium chloride solution (3.5 mL) of the same concentration was added and allowed to stand for 5 minutes. The gel was washed twice with 5 mL of physiological saline to obtain a chemically crosslinked alginic acid gel encapsulating fluorescein isothiocyanate-dextran. 19.5 mL of physiological saline was added to this gel, shaken at 37 °C, and the aqueous solution was recovered over time, and PBS equal to the recovered amount was replenished. After the test, 10 μL of alginate lyase (Nippon Gene, 319-08261) was added to the test solution, and shaken at 37 °C for more than 3 hours to completely disintegrate the gel, and the aqueous solution was recovered. The dextran concentration in the recovered aqueous solution was measured by fluorescence quantification (excitation light: 485 nm, fluorescence: 535 nm), and the value obtained by dividing the amount of dextran at each time point by the amount of dextran after the end of the test was expressed as a percentage, and the obtained value was used as the transmittance.

[1189] The Figure 10 results were obtained. The transmittance after 3 hours was about 5 to about 10%. And the transmittance after 24 hours was about 26 to about 37%.

[1190] [Biocompatibility evaluation]

[1191] Prepare the alginate derivatives (AL-EX-2), (AL-EX-4), (AL-EX-5), (AL-EX-6), (AL-EX-7.1) obtained in (Example 2), (Example 4), (Example 5), (Example 6), (Example 7) and (AL-EX-10.1), (AL-EX-11.1), (AL-EX-12.1) and (AL-EX-13.1) prepared by the same method as (Example 10), (Example 11), (Example 12), (Example 13) to form a PBS solution with a concentration of 1.0% by weight to obtain an aqueous alginate solution.

[1192] Filter and sterilize it using Minisart highflow (Sartorius, 16532GUK) to obtain alginate PBS solutions (2), (4), (5), (6), (7), (10), (11), (12) and (13) with 1.0% crosslinking groups introduced.

[1193] In HeLa cells cultured for 1 day after seeding in a 96-well plate at a cell concentration of 5×10^3 cells / well, add the alginate PBS solution with 1.0% crosslinking groups introduced in combinations of (2) or (5), and (7), (10), (11), (12) or (13), and combinations of (4) or (6) and (7) to a final concentration of 0.1%. After culturing for 1 day, evaluate the ATP activity as an index of cytotoxicity using CellTiter-Glo luminescent cell viability assay (Promega, G7571).

[1194] Obtain Figure 7 the results. In all the aforementioned crosslinked alginate gels, even when compared with PBS and alginate (A-2) as controls, the same level of ATP activity can be confirmed, indicating that there is no cytotoxicity in the crosslinked alginate gels, and it shows that the alginate structure formed by the Michael reaction has biocompatibility.

[1195] [Biocompatibility evaluation (2)]

[1196] Prepare the alginate derivatives (AL-EX-2), (AL-EX-5), (AL-EX-7.1) obtained in (Example 2), (Example 5) and (Example 7) and (AL-EX-14.1), (AL-EX-15.1) and (AL-EX-16.1) prepared by the same method as (Example 14), (Example 15), (Example 16) to form a PBS solution with a concentration of 1.0% by weight to obtain an aqueous alginate solution.

[1197] Filter sterilize it using Minisart highflow (Sartorius, 16532GUK) to obtain alginate PBS solutions (2), (5), (7), (14), (15), and (16) with 1.0% crosslinking groups introduced.

[1198] In HeLa cells obtained by culturing for 1 day after seeding in a 96-well plate at a cell concentration of 5×10^3 cells / well, add the alginate PBS solution with 1.0% crosslinking groups introduced in combinations of (2) or (5), and (7), (14), (15), or (16) to a final concentration of 0.1%. After culturing for 1 day, evaluate the ATP activity as an index of cytotoxicity using CellTiter-Glo luminescent cell viability assay (Promega, G7571).

[1199] Obtain Figure 11 the results. In all of the aforementioned crosslinked alginate gels, even when compared with PBS and alginate (A-2) used as controls, the same level of ATP activity was confirmed. This indicates that there is no cytotoxicity in the crosslinked alginate gels, showing that the alginate structure formed by the Michael reaction has biocompatibility.

Claims

1. Crosslinked alginic acid, which is obtained by carrying out a crosslinking reaction using an alginic acid derivative represented by the following formula (I) and an alginic acid derivative represented by the following formula (II); [Chemical formula 132] In formula (I), (ALG) represents alginic acid; -NHCO- represents an amide bond obtained from any carboxyl group of alginic acid; -L 1 - is a linking group selected from the following partial structural formulas, and in each formula, the outside of the dotted lines at both ends is not included, [Chemical formula 133] The methylene -CH in formulas (L1-1) to (L1-4) 2 - hydrogen atoms can be selected from oxo group =O, halogen atoms, hydroxy groups, C 1～6 alkyl groups, hydroxy C 1～6 alkyl groups, mercapto C 1～6 alkyl groups, C 1～6 alkylthio C 1～6 alkyl groups, -COOH group, -COOM group, -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl group, (R a R b N)C(=O)-C 1～6 alkyl group, guanidino C 1～6 alkyl group, C 7～16 arylalkyl group, hydroxy C 6～10 aryl C 1～6 alkyl group or heteroaryl C 1～6 alkyl group and be substituted by multiple groups among them; Wherein, M is Li, Na, K or 1 / 2Ca, and the foregoing -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl, and R a and R b are each independently a group selected from a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl, In formula (L1-1) to formula (L1-4), the two hydrogen atoms of the same methylene group -CH 2 - are substituted with C 1～6 alkyl groups, and these alkyl groups may be bonded to each other to form a C 3～8 cycloalkyl ring; The -NH- group in formula (L1-3) and formula (L1-4) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom; n is an integer from 1 to 18; m is an integer from 1 to 9; j is an integer from 0 to 9; [Chemical formula 134] In formula (II), (ALG) represents alginic acid; -NHCO- represents an amide bond obtained from any carboxyl group of alginic acid; -L 2 - is a linking group selected from the following partial structural formulas, and in each formula, the outside of the dotted lines at both ends is not included, [Chemical formula 135] The methylene -CH in formulas (L2-1) to (L2-6) 2 - hydrogen atoms can be selected from oxo group =O, halogen atoms, hydroxyl group, C 1～6 alkyl, hydroxy C 1～6 alkyl, mercapto C 1～6 alkyl, C 1～6 alkylthio C 1～6 alkyl, -COOH group, -COOM group, -COO(C 1～6 alkyl) group, -NR a R b group, (R a R b N)-C 1～6 alkyl, (R a R b N)C(=O)-C 1～6 alkyl, guanidino C 1～6 alkyl, C 7～16 aralkyl, hydroxy C 6～10 aryl C 1～6 alkyl or heteroaryl C 1～6 alkyl; wherein, M is Li, Na, K or 1 / 2Ca, and the foregoing -NR a R b group, (R a R b N)-C 1～6 alkyl or (R a R b N)C(=O)-C 1～6 alkyl in which R a and R b are each independently selected from a group consisting of a hydrogen atom, C 1～6 alkyl, C 2～7 alkanoyl or C 1～6 alkylsulfonyl. In formulae (L2-1) to (L2-6), when two hydrogen atoms of the same methylene group -CH 2 2 - are substituted with C 1～6 1～6 alkyl groups, the alkyl groups may be bonded to each other to form a C 3～8 3～8 cycloalkyl ring; The -NH- group in formula (L2-3) to formula (L2-6) may form a non-aromatic heterocycle together with the substituent bonded to the adjacent carbon atom; m2 is an integer from 1 to 9; n2 is an integer from 1 to 18; m3 is an integer from 1 to 10; n3 is an integer from 1 to 10; j2 is an integer from 0 to 9.

2. The crosslinked alginic acid according to claim 1, Wherein, The chemical crosslinking of the crosslinked alginic acid is a structure of the following formula (LK-1): [Chemical formula 139] In formula (LK-1), -CONH- and -NHCO- at both ends represent amide bonds obtained from any carboxyl group of alginic acid; -L 1 - and -L 2 - has the same definition as that in Claim 1.

3. The crosslinked alginic acid according to claim 1 or 2, which has biocompatibility.

4. A method for preparing the crosslinked alginic acid according to claim 1, which comprises adding a solution of the alginic acid derivative represented by formula (I) to a solution of the alginic acid derivative represented by formula (II) to carry out a crosslinking reaction.

5. A method for preparing the crosslinked alginic acid according to claim 1, which comprises adding a solution of the alginic acid derivative represented by formula (II) to a solution of the alginic acid derivative represented by formula (I) to carry out a crosslinking reaction.

6. A method for preparing the crosslinked alginic acid according to claim 1, Wherein, The chemical crosslinking formed by carrying out a Michael addition reaction using the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) is a structure of the following formula (LK-1): [Chemical formula 140] In formula (LK-1), the -CONH- and -NHCO- at both ends represent amide bonds obtained from any carboxyl group of alginic acid; -L 1 -, and -L 2 - has the same definition as that in claim 1.

7. A crosslinked alginic acid structure, which is obtained by carrying out a crosslinking reaction on a gel obtained by dropping a solution of the alginic acid derivative represented by formula (I) into a solution containing a divalent metal ion in a solution of the alginic acid derivative represented by formula (II), and as crosslinking, includes ionic crosslinking partially formed by the divalent metal ion and chemical crosslinking formed by a Michael addition reaction, In formula (I), -L 1 - is the same as defined in claim 1; in formula (II), -L 2 - is the same as defined in claim 1.

8. The crosslinked alginic acid structure according to claim 7, Wherein, The chemical crosslinking formed by carrying out a Michael addition reaction using the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) is a structure of the following formula (LK-1): [Chemical formula 141] In formula (LK-1), -CONH- and -NHCO- at both ends represent amide bonds obtained from any carboxyl group of alginic acid; -L 1 - and -L 2 - have the same definition as that in claim 1.

9. The crosslinked alginic acid structure according to claim 7 or 8, which is a fibrous structure, a bead or a gel.

10. The crosslinked alginic acid structure according to claim 7 or 8, which is a fiber or a near-spherical gel.

11. The crosslinked alginic acid structure according to claim 7 or 8, which has biocompatibility.

12. A crosslinked alginic acid structure, which is obtained by subjecting a gel obtained by dropping a solution of an alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of the alginic acid derivative represented by formula (I). As the crosslinking, it includes ionic crosslinking partially formed by the divalent metal ion and chemical crosslinking formed by a Michael addition reaction. In formula (II), -L 2 - is the same as defined in claim 1; in formula (I), -L 1 - is the same as defined in claim 1.

13. The crosslinked alginic acid structure according to claim 12, wherein the chemical crosslinking formed by the Michael addition reaction using the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) has the structure of the following formula (LK-1): [Chemical formula 141] In formula (LK-1), -CONH- and -NHCO- at both ends represent amide bonds obtained from any carboxyl group of alginic acid; -L 1 -and -L 2 - is the same as the definition in claim 1.

14. The crosslinked alginic acid structure according to claim 12 or 13, which is a fibrous structure, a bead or a gel.

15. The crosslinked alginic acid structure according to claim 12 or 13, which is a fiber or a near-spherical gel.

16. The crosslinked alginic acid structure according to claim 12 or 13, which has biocompatibility.

17. A crosslinked alginic acid structure, which is obtained by dropping a solution of a composition containing the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion. As the crosslinking, it includes ionic crosslinking partially formed by the divalent metal ion and chemical crosslinking formed by a Michael addition reaction. In formula (I), -L 1 - is the same as defined in claim 1; in formula (II), -L 2 - is the same as defined in claim 1.

18. The crosslinked alginic acid structure according to claim 17, wherein the chemical crosslinking formed by the Michael addition reaction using the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) has the structure of the following formula (LK-1): [Chemical formula 141] In formula (LK-1), -CONH- and -NHCO- at both ends represent amide bonds obtained from any carboxyl group of alginic acid; -L 1 -, and -L 2 - has the same definition as that in claim 1.

19. The crosslinked alginic acid structure according to claim 17 or 18, which is a fibrous structure, a bead or a gel.

20. The crosslinked alginic acid structure according to claim 17 or 18, which is a fiber or a near-spherical gel.

21. The crosslinked alginic acid structure according to claim 17 or 18, which has biocompatibility.

22. A medical material, which contains the crosslinked alginic acid structure according to any one of claims 7 to 21.

23. The medical material according to claim 22, which is a fibrous structure, a bead or a gel.

24. The medical material according to claim 22, which is a fiber or a near-spherical gel.

25. A composition, which contains the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II). In formula (I), -L 1 - is the same as defined in claim 1; in formula (II), -L 2 - is the same as defined in claim 1.

26. A method for preparing a crosslinked alginic acid structure, which includes dropping a solution of a composition containing the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion. In formula (I), -L 1 - is the same as defined in claim 1; in formula (II), -L 2 - is the same as defined in claim 1.

27. A method for preparing a crosslinked alginic acid structure, which includes: By subjecting a gel obtained by dropping a solution of an alginic acid derivative represented by formula (I) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of an alginic acid derivative represented by formula (II), a crosslinked alginic acid structure is obtained which contains an ionic crosslink partially formed by a divalent metal ion and a chemical crosslink formed by a Michael addition reaction as crosslinks. In formula (I), -L 1 - is the same as defined in claim 1; in formula (II), -L 2 - is the same as defined in claim 1.

28. A method for preparing a crosslinked alginic acid structure, which comprises: By subjecting a gel obtained by dropping a solution of an alginic acid derivative represented by formula (II) into a solution containing a divalent metal ion to a crosslinking reaction in a solution of an alginic acid derivative represented by formula (I), a crosslinked alginic acid structure is obtained which contains an ionic crosslink partially formed by a divalent metal ion and a chemical crosslink formed by a Michael addition reaction as crosslinks. In formula (II), -L 2 - is the same as defined in claim 1; in formula (I), -L 1 - is the same as defined in claim 1.

29. The method for preparing a crosslinked alginic acid structure according to any one of claims 26 to 28, wherein the chemical crosslink formed by a Michael addition reaction using the alginic acid derivative represented by formula (I) and the alginic acid derivative represented by formula (II) has the structure of the following formula (LK-1): [Chemical 142] In formula (LK-1), the -CONH- and -NHCO- at both ends represent amide bonds obtained from any carboxyl group of alginic acid; -L 1 -, and -L 2 - have the same definition as in claim 1.

30. A crosslinked alginic acid structure, which is obtained by subjecting an alginic acid derivative represented by formula (I) and an alginic acid derivative represented by formula (II) to ionic crosslinking with a divalent metal ion and chemical crosslinking by a Michael addition reaction, and has a retention property of the content. In formula (I), -L 1 - is the same as defined in claim 1; in formula (II), -L 2 - is the same as defined in claim 1.

31. The crosslinked alginic acid structure according to claim 30, which has biocompatibility.

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