Hydrogel

Abstract

A hydrogel containing a compound represented by formula (1) and a compound represented by formula (2), wherein at least one substrate site of the compound represented by formula (1) and at least one ligand site of the compound represented by formula (2) are coordinately bonded to each other. The symbols each have the same meaning as set forth in the description.

Description

Hydrogel

[0001] The present disclosure relates to hydrogels.

[0002] Injectable gels have the property of gelling when two liquids are mixed. For example, Non-Patent Document 1 describes that placing drug-loaded nanodisks in a solution causes the drug to be slowly released after gelling in the body. Non-Patent Document 2 describes that cartilage is repaired by forming a cross-linked gel structure after injection into a test subject.

[0003] Z. Cimen et al., ACS Appl. Polym. Mater. 2021, 3, 3504.S. Li et al., ACS Biomater. Sci. Eng. 2023, 9, 2625.

[0004] However, it has been difficult to control the lifespan (solation time) of hydrogels using conventional techniques. The present disclosure aims to provide a hydrogel that can be administered topically.

[0005] The present disclosure provides the following [1] to

[21] : [1] A hydrogel comprising a compound represented by formula (1) and a compound represented by formula (2), wherein at least one substrate moiety of the compound represented by formula (1) and at least one ligand moiety of the compound represented by formula (2) are coordinately bonded. [In formula (1), R Ａ１ is the maternal site; X １ are each independently a single bond or a divalent or higher valent organic group; R Ｂ１ is the substrate site; R Ｂ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms; α1 is an integer of 1 or more; α2 is an integer of 0 or more; the sum of α1 and α2 is R Ａ１ γ1 is an integer of 1 or more; γ2 is an integer of 1 or more; In formula (2), R Ａ２ is the maternal site; X ２are each independently a single bond or a divalent or higher valent organic group; R Ｃ１ is the ligand site; R Ｃ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms; β1 is an integer of 1 or more; β2 is an integer of 0 or more; the sum of β1 and β2 is R Ａ２ δ1 is an integer of 1 or more; δ2 is an integer of 1 or more.] [2] R Ａ１ and R Ａ２ The hydrogel according to [1], wherein R each independently comprise a polyalkylene glycol backbone, an α-poly-L-glutamic acid backbone, an α-poly-D-glutamic acid backbone, a γ-poly-L-glutamic acid backbone, a γ-poly-D-glutamic acid backbone, an α-poly-L-lysine backbone, an α-poly-D-lysine backbone, a dextran backbone, an alginic acid backbone, a polyvinyl alcohol backbone, a polyacrylic acid backbone, or a polymethacrylic acid backbone. [3] The hydrogel according to [2], wherein the polyalkylene glycol backbone is a polyethylene glycol backbone. [4] R Ａ１ is -C(C ｃ１ H ２ｃ１ -O-(C ａ１ H ２ａ１ O) ｂ１１ -) α１ c1 is (C ｃ１ H ２ｃ１ a1 is independently an integer of 1 to 6 for each (C ａ１ H ２ａ１ The hydrogel according to any one of [1] to [3], wherein each of R 11 and R 21 is independently an integer of 1 to 6; b11 is an integer of 1 to 10,000; and α1 is an integer of 2 to 4. [5] R Ａ２ is -C(C ｆ１ H ２ｆ１ -O-(C ｄ１ H ２ｄ１ O) ｅ１１ -) β１ f1 is (C ｆ１ H ２ｆ１ ) units are each independently an integer of 1 to 6; d1 is (C ｄ１ H２ｄ１ ) units are each independently an integer of 1 to 6; e11 is an integer of 1 to 10,000; and β1 is an integer of 2 to 4. [6] R Ｂ１ are each independently —O—PO(OH) ２ [7] The hydrogel according to any one of [1] to [5], wherein R is a group containing a Ｂ１ are each independently —O—PO(OH) ２ , -NHCH(C(=O)OH)(CH ２ ) ｍ４１ —O—PO(OH) ２ (wherein m41 is an integer of 1 to 3), —NHCH(C(═O)OH)(CH ２ ) ｍ４２ -CH(-O-PO(OH) ２ )-CH ３ (wherein m42 is an integer of 0 to 3), or —NHCH(C(═O)OH)(CH ２ ) ｍ４３ -Ph(-O-PO(OH) ２ ) (wherein m43 is an integer of 1 to 3). [8] R Ｃ１ [9] The hydrogel according to any one of [1] to [7], wherein R is independently a group containing a metal atom.

[10] The hydrogel according to [8], wherein the metal atom is a divalent metal atom.

[11] Ｃ１ has the following structure: (wherein * is a bond; m2 is an integer from 1 to 6; M ２＋

[11] The hydrogel according to any one of [1] to [9], wherein X is a divalent metal atom. １ and X ２ are each independently -(CH ２ ) ｍ１１The hydrogel according to any one of [1] to

[10] , wherein m11 is —C(═O)— and m11 is an integer of 1 to 10.

[12] A composition, in which at least a part of the coordinate bonds between the substrate moiety and the ligand moiety are decoupled by adding a first enzyme to the hydrogel according to any one of [1] to

[11] .

[13] The composition according to

[12] , in which the first enzyme is a phosphatase.

[14] The composition according to

[12] or

[13] , which gels upon addition of a second enzyme.

[15] The composition according to

[14] , in which the second enzyme is a kinase.

[16] A compound represented by formula (1): R Ａ１ is the maternal site; X １ are each independently a single bond or a divalent or higher valent organic group; R Ｂ１ is the substrate site; R Ｂ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms; α1 is an integer of 1 or more; α2 is an integer of 0 or more; the sum of α1 and α2 is R Ａ１

[17] A compound in which R is a valence of γ1; γ2 is an integer of 1 or more; Ａ１ is a polyethylene glycol backbone (-(C ａ１ H ２ａ１ O) ｂ１１ a1 is an integer from 1 to 6; b11 is an integer from 1 to 10,000; X １ is -(CH ２ ) ｍ１１ -C(=O)-; m11 is an integer of 1 to 10; R Ｂ１ are each independently —NHCH(C(═O)OH)—(CH ２ ) ｍ１ —O—PO(OH) ２

[18] The compound according to

[16] , wherein m1 is an integer of 1 to 3, α1 is 4, and α2 is 0.

[18] The compound according to

[16] , R Ａ２ is the maternal site; X ２are each independently a single bond or a divalent or higher valent organic group; R Ｃ１ is the ligand site; R Ｃ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms; β1 is an integer of 1 or more; β2 is an integer of 0 or more; the sum of β1 and β2 is R Ａ２

[19] A compound in which R is a valence of 1 or more; δ1 is an integer of 1 or more; and δ2 is an integer of 1 or more. Ａ２ is a polyethylene glycol backbone (-(C ｄ１ H ２ｄ１ O) ｅ１１ -); d1 is an integer from 1 to 6; e11 is an integer from 1 to 10,000; X ２ is -(CH ２ ) ｍ１１ -C(=O)-; m11 is an integer of 1 to 10; R Ｃ１ each independently have the following structure: The compound according to

[18] , wherein * is a binding moiety; and m2 is an integer of 1 to 6.

[20] A composition comprising the compound according to

[18] or

[19] and a first enzyme.

[21] The composition according to

[20] , further comprising a compound of formula (1): A composition comprising a compound represented by the formula: Ａ１ is the maternal site; X １ are each independently a single bond or a divalent or higher valent organic group; R Ｂ１ is the substrate site; R Ｂ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms; α1 is an integer of 1 or more; α2 is an integer of 0 or more; the sum of α1 and α2 is R Ａ１ γ1 is an integer of 1 or more; and γ2 is an integer of 1 or more.

[0006] According to the present disclosure, a hydrogel that can be administered topically can be provided.

[0007] 1 shows the relationship between the time after mixing and the results of the values ​​of storage modulus (G') and loss modulus (G'') in Example 1-1. The values ​​of storage modulus (G') and loss modulus (G'') in Examples 1-2 to 1-3 are shown. The relationship between the time after mixing and the state of a system without phosphatase added (0 unit of phosphatase) and a system with phosphatase added (1 unit of phosphatase) is shown. The relationship between the time after mixing and the state of the hydrogel is shown. The relationship between the standing time and leakage rate (%) in Example 4 and Comparative Example 1 is shown. The relationship between the standing time and leakage rate (%) in Examples 4 and 5 is shown. The relationship between the concentration of phosphatase and leakage rate (%) is shown. The leakage rate (%) in Examples 8-1, 8-2, and 8-3 is shown.

[0008] As used herein, the term "organic group" refers to a group containing carbon. The organic group is not particularly limited, but may be a hydrocarbon group. The divalent organic group is not particularly limited, but may be a divalent group obtained by further eliminating one hydrogen atom from a hydrocarbon group.

[0009] As used herein, the term "hydrocarbon group" refers to a group containing carbon and hydrogen, resulting from the elimination of one hydrogen atom from a hydrocarbon. Such hydrocarbon groups are not particularly limited, but include hydrocarbon groups having 1 to 10 carbon atoms, which may be substituted with one or more substituents, such as aliphatic hydrocarbon groups and aromatic hydrocarbon groups. The above-mentioned "aliphatic hydrocarbon group" may be linear, branched, or cyclic, and may be saturated or unsaturated. The hydrocarbon group may also contain one or more ring structures.

[0010] First Embodiment [Hydrogel] The hydrogel of the present disclosure contains a compound represented by formula (1) and a compound represented by formula (2).

[0011] The hydrogel of the present disclosure forms a gel through a coordinate bond between the substrate moiety contained in formula (1) and the ligand moiety contained in formula (2). As will be described later, the hydrogel can react with an enzyme to form a sol. This sol-gel reaction can enable the local administration of drugs and the like.

[0012] [Formula (1)] R Ａ１ is a parent moiety, and a commonly used group can be used. Ａ１ Preferably, the polymer contains a plurality of polyalkylene glycol backbones, α-poly-L-glutamic acid backbones, α-poly-D-glutamic acid backbones, γ-poly-L-glutamic acid backbones, γ-poly-D-glutamic acid backbones, α-poly-L-lysine backbones, α-poly-D-lysine backbones, dextran backbones, alginic acid backbones, polyvinyl alcohol backbones, polyacrylic acid backbones, or polymethacrylic acid backbones. Note that the term "backbones" refers to the contiguous presence of specific repeating units. The "specific repeating units" may be of one type or of multiple types (e.g., two, three, four, etc.).

[0013] Each skeleton has a repeating unit, and the number of repeating units is, for example, an integer from 1 to 10,000, preferably an integer from 1 to 1,000, and more preferably an integer from 40 to 1,000. The number of repeating units may be an integer from 1 to 800, or may be an integer from 20 to 800.

[0014] The terminal portion of the backbone main chain is a linker portion (e.g., X １ or X ２ The main chain refers to a chain in which repeating portions are linked in a skeleton. The polymerization-terminating portion may be a portion where polymerization does not proceed, and is not particularly limited, but may be, for example, an H atom, an OH group, an NH ２ group or NH(CH ２ ) ｂ２０ CH ３ group (b20 is an integer of 0 to 10).

[0015] R Ａ１ In the formula (I), the polyalkylene glycol backbone is preferably -(C ａ１ H２ａ１ O) ｂ１１ a1 is an integer of 1 to 6, preferably 2 to 4, and more preferably 2. In other words, the polyalkylene glycol skeleton is preferably a polyethylene glycol skeleton. b11 is an integer of 1 to 10,000, preferably an integer of 1 to 1,000, and more preferably an integer of 40 to 1,000, and may be, for example, an integer of 50 to 1,000. In addition, when the right side of the polyalkylene glycol skeleton is a linker X １ Combine with.

[0016] R Ａ１ In the formula (I), the polyglutamic acid backbone is preferably —(NHCH(C ＊ (=O))CH ２ CH ２ C(=O) ｂ１２ The carbon atom (atom marked with *) of the carbonyl group (C(═O)) of the polyglutamic acid backbone contains the linker X １ It is to be noted that R Ａ１ In the whole, at least one * is a linker X １ The other * may be bonded to an OH group. Ａ１ In the whole, at least one * is a linker X １ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ１ b12 is an integer of 1 to 10,000, preferably an integer of 1 to 1,000, more preferably an integer of 1 to 800, and particularly preferably an integer of 20 to 800. R Ａ１ may have a polymerization terminating moiety at the end, for example, NH ２ CH(C(=O)OH)CH ２ CH ２ C(=O)-(NHCH(C ＊ (=O))CH ２ CH ２ C(=O) ｂ１２ -NHCH(C(=O)OH)CH ２ CH ２ It may be represented by C(=O)OH.

[0017] R Ａ１ In the formula (I), the lysine backbone may preferably have the following structure: Ａ１ In the whole, at least one * is a linker X １ The other * may be bonded to a H atom. Ａ１ In the whole, at least one * is a linker X １ All * in one unit enclosed in brackets may be bonded to H atoms as long as they are bonded to R. In this case, the H atoms are Ａ１ Included in. b13 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. Ａ１ may have a polymerization terminating moiety at the end, and may be represented by, for example, the following formula: R ｂ３ is not particularly limited, but may be, for example, an OH group, NH ２ group or NH(CH ２ ) ｂ２０ CH ３ b20 is an integer of 0 to 10, preferably an integer of 2 to 5.

[0018] R Ａ１ In the formula, the dextran backbone may have the following structure: Ａ１ In the whole, at least one * is a linker X １ The other * may be bonded to an OH group. Ａ１ In the whole, at least one * is a linker X １ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ１ Included in. In the formula, b14 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. Ａ１ may have a polymerization terminating moiety at the end, and may be represented by, for example, the following formula:

[0019] R Ａ１ In the above, the alginate backbone may have the following structure: Ａ１In the whole, at least one * is a linker X １ The other * may be bonded to an OH group. Ａ１ In the whole, at least one * is a linker X １ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ１ Included in. In the formula, b15 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000, and b16 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. The sum of b15 and b16 may be an integer of 1 to 10,000, or may be an integer of 1 to 1,000. R Ａ１ may have a polymerization terminating moiety at the end, and may be represented by, for example, the following formula:

[0020] R Ａ１ In the formula, the polyvinyl alcohol backbone is -[C ＊ HCH ２ ] ｂ１７ - may have R Ａ１ In the whole, at least one * is a linker X １ The other * may be bonded to an OH group. Ａ１ In the whole, at least one * is a linker X １ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ１ b17 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. Ａ１ may have a polymerization terminating moiety at the end, for example, H—[C ＊ HCH ２ ] ｂ１７ It may be represented by —COOH.

[0021] R Ａ１ In the formula, the polyacrylic acid backbone is -[CH ２ CH(C ＊ (=O)) ｂ１８ - may have R Ａ１In the whole, at least one * is a linker X １ The other * may be bonded to an OH group. Ａ１ In the whole, at least one * is a linker X １ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ１ b18 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. Ａ１ may have a polymerization terminating moiety at the end.

[0022] R Ａ１ In the formula, the polymethacrylic acid backbone is -[CH ２ CCH ３ (C ＊ (=O)) ｂ１９ - may have R Ａ１ In the whole, at least one * is a linker X １ The other * may be bonded to an OH group. Ａ１ In the whole, at least one * is a linker X １ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ１ b19 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. Ａ１ may have a polymerization terminating moiety at the end.

[0023] When used on animals or other test subjects, R Ａ１ is preferably a structure that has little effect on the test subject, for example, R Ａ１ As the backbone, a polyalkylene glycol backbone or a polyglutamic acid backbone can be used.

[0024] R Ａ１ can bind to one or more substrate moieties. That is, α1 is an integer of 1 or more, and α2 is an integer of 0 or more. However, the sum of α1 and α2 is not greater than R Ａ１ is the valence of

[0025] In one embodiment, α1 is an integer of 1 or more, and α2 is an integer of 0 or more.

[0026] In one embodiment, α1 is an integer of 2 or more, and α2 is an integer of 0 or more.

[0027] For example, R Ａ１ When is tetravalent, preferably, α1 is an integer of 2 to 4, and α2 is an integer of 0 to 2, and more preferably, α1 is 4.

[0028] For example, when α1 is 4 and α2 is 0, equation (1) is expressed as follows:

[0029] R Ａ１ For example, -C(C ｃ１ H ２ｃ１ -O-(C ａ１ H ２ａ１ O) ｂ１１ -) α１ In the formula, c1 is represented by (C ｃ１ H ２ｃ１ a1 is independently an integer of 1 to 6, preferably an integer of 1 to 4, more preferably 1 for each (C ａ１ H ２ａ１ ) units are each independently an integer of 1 to 6, preferably an integer of 2 to 4, and more preferably 2; b11 is an integer of 1 to 10,000, preferably an integer of 1 to 1,000, and more preferably an integer of 40 to 1,000, for example, an integer of 50 to 1,000; α1 is an integer of 2 to 4, preferably 3 or 4, and more preferably 4.

[0030] R Ｂ１ is the substrate site. Ｂ１ are each independently preferably —O—PO(OH) ２ It is a group containing a group.

[0031] R Ｂ１ is more preferably —O—PO(OH) ２ , -NHCH(C(=O)OH)(CH ２ ) ｍ４１ —O—PO(OH) ２ (wherein m41 is an integer of 1 to 3, preferably 1), —NHCH(C(═O)OH)(CH２ ) ｍ４２ -CH(-O-PO(OH) ２ )-CH ３ (wherein m42 is an integer of 0 to 3, preferably 0), —NHCH(C(═O)OH)(CH ２ ) ｍ４３ -Ph(-O-PO(OH) ２ ) (wherein m43 is an integer of 1 to 3, preferably 1, and Ph is an o-, m-, or p-phenylene group, preferably a p-phenylene group).

[0032] R Ｂ１ Examples of the alkyl group include -O-PO(OH) ２ , -NHCH(C(=O)OH)CH ２ O-PO(OH) ２ , -NHCH(C(=O)OH)CH(-O-PO(OH) ２ ) CH ３ , Examples include:

[0033] R Ｂ２ is a group that does not have a substrate moiety. Ｂ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms. Ｂ２ is, for example, -(CH ２ ) ｎ３１ -OH, -(CH ２ ) ｎ３１ -COOH, -(CH ２ ) ｎ３１ -H or -(CH ２ ) ｎ３１ -CH ３ Each n31 is independently an integer of 0 to 6.

[0034] X １ is a linker moiety and is a single bond or a divalent or higher organic group. １ is the maternal part R Ａ１ and substrate site R Ｂ１ or the parent site R Ａ１ and R Ｂ２ and are linked together.

[0035] X １In one embodiment, X is a single bond. １ In one embodiment, is a divalent or higher organic group.

[0036] X １ The divalent or higher organic group in the formula (I) contains, for example, at least one selected from the group consisting of alkylene, -C(=O)-, -C(=O)-NH-, -NH-C(=O)-, and phenylene. The hydrogen atom of the divalent or higher organic group may be substituted.

[0037] X １ are preferably each independently -X １１ -X １２ It is represented by -. １１ R on the side Ａ１ To, X １２ R on the side Ｂ１ or R Ｂ２ Connect to the respective

[0038] X １１ has any one of the following structures: In the following formulas, in **, R Ａ１ In ##, X １２ are connected to the respective single bonds; ＊＊ (CH ２ ) ｍ１１ -C ＃＃ (═O) (x1) [wherein: m11 is each independently an integer of 1 to 10, preferably an integer of 3 to 7, for example, 5]; or Formula (x2): [wherein: m12 is an integer of 1 to 10, preferably 2; m13 is an integer of 1 to 10, preferably 2; and m14 is an integer of 1 to 10, preferably 2.]

[0039] X １２ has one of the following structures: In the following structures, # represents X １１ , * is R Ｂ１ or R Ｂ２ A single bond is connected to each of the following: [wherein m15 is an integer of 1 to 1,000, preferably an integer of 1 to 50]; Formula (z2): [wherein m16 is an integer of 1 to 1,000, preferably an integer of 1 to 50]; Formula (z3): [wherein m17 is an integer of 1 to 1,000, preferably an integer of 1 to 50]; Formula (z4): [wherein: x is an integer of 1 to 1,000, preferably an integer of 1 to 800, more preferably an integer of 1 to 50; y is an integer of 1 to 1,000, preferably an integer of 1 to 800, more preferably an integer of 1 to 50; the order of occurrence of the repeating units enclosed in parentheses around x and y is arbitrary in the formula; m18 is an integer of 1 to 6, preferably an integer of 2 to 5, more preferably 2; m19 is an integer of 1 to 6, preferably an integer of 1 to 5, more preferably 1.] (In another example, x may be an integer of 5 to 400, y may be an integer of 5 to 400, and the sum of x and y may be an integer of 5 to 400.) Formula (z5): [wherein: x is an integer of 1 to 1,000, preferably an integer of 1 to 800, more preferably an integer of 1 to 50; y is an integer of 1 to 1,000, preferably an integer of 1 to 800, more preferably an integer of 1 to 50; the order of the repeating units enclosed in parentheses around x and y is arbitrary in the formula; m20 is an integer of 1 to 6, preferably an integer of 2 to 5, more preferably 2; m21 is an integer of 1 to 6, preferably an integer of 1 to 5, more preferably 1.]; Formula (z6): [wherein: x is an integer of 1 to 1,000, preferably an integer of 1 to 800, more preferably an integer of 1 to 50; y is an integer of 1 to 1,000, preferably an integer of 1 to 800, more preferably an integer of 1 to 50; the order of the repeating units enclosed in parentheses with x and y is arbitrary in the formula; and m22 is an integer of 1 to 6, preferably an integer of 2 to 5, more preferably 3.]; Formula (z7): [In the formula: m23 is independently an integer of 1 to 6, preferably an integer of 2 to 4, for example, 3; m24 is independently an integer of 1 to 6, preferably an integer of 2 to 4, for example, 2; and m25 is independently an integer of 1 to 1,000, preferably an integer of 10 to 50.]

[0040] Preferably, X １１ and X １２ and X are not both single bonds. １１ is represented by formula (x1) or (x2), and X １２ is a single bond or is represented by any one of formulas (z1) to (z7).

[0041] -X １２ -R Ｂ１ Examples of the groups include the following: １１ Binds to (HO) ２ OP-OCH ２ CH(C(=O)OH)NH-C(=O)(CH ２ ) ｍ１１ ＃ ,

[0042] γ1 is a linker X １ R bonded to Ｂ１ γ1 is an integer of 1 or more. The value of γ1 is the number of linkers X １ The upper limit of γ1 can vary depending on the structure of the linker X １ The valence of the atom can be −1.

[0043] R Ａ１ When is a polyalkylene glycol backbone, γ1 can be 1 or more for the entire polyalkylene glycol backbone.

[0044] R Ａ１ is a glutamic acid backbone, γ1 can be 1 or more for the entire glutamic acid backbone. Ａ１ can be b12 or less for the entire glutamic acid backbone.

[0045] R Ａ１ is a lysine backbone, γ1 can be 1 or greater across the lysine backbone. γ1 can be b13 or less across the lysine backbone.

[0046] R Ａ１ When b14 is a dextran backbone, γ1 can be 1 or more relative to the entire dextran backbone. γ1 can be 3×b14 or less relative to the entire dextran backbone. γ1 can be, for example, b14 or less.

[0047] R Ａ１ is an alginate backbone, γ1 can be 1 or greater for the entire alginate backbone. γ1 can be 3×(b15+b16) or less for the entire alginate backbone. γ1 may be, for example, (b15+b16) or less.

[0048] R Ａ１ is a polyvinyl alcohol backbone, γ1 can be 1 or more for the entire polyvinyl alcohol backbone. γ1 can be b17 or less for the entire polyvinyl alcohol backbone.

[0049] R Ａ１ is a polyacrylic acid backbone, γ1 can be 1 or more relative to the entire polyacrylic acid backbone. γ1 can be b18 or less relative to the entire polyacrylic acid backbone.

[0050] R Ａ１ When b is a polymethacrylic acid backbone, γ1 can be 1 or more for the entire polymethacrylic acid backbone. γ1 can be b19 or less for the entire polymethacrylic acid backbone.

[0051] γ2 is a linker X １ R bonded to Ｂ２ γ2 is an integer of 1 or more. The value of γ2 is the number of linkers X １ The upper limit of γ2 can vary depending on the structure of the linker X １ The valence of the atom can be −1.

[0052] γ2 can take the same numerical range as γ1.

[0053] In addition, -X １ (-R Ｂ２ )γ２ is R in the structure Ｂ１ This refers to a structure that does not contain any of the above.

[0054] In one embodiment, X １２ is a single bond, R Ｂ１ is -NHCH(C(=O)OH)(CH ２ ) ｍ４１ —O—PO(OH) ２ (wherein m41 is an integer of 1 to 3, preferably 1), —NHCH(C(═O)OH)(CH ２ ) ｍ４２ CH(-O-PO(OH) ２ ) CH ３ (wherein m42 is an integer of 0 to 3, preferably 0), or —NHCH(C(═O)OH)(CH ２ ) ｍ４３ -Ph(-O-PO(OH) ２ ) (wherein m43 is an integer of 1 to 3, preferably 1, and Ph is o-, m-, or p-phenylene, preferably p-phenylene).

[0055] In one embodiment, X １２ is expressed by the formula (z4): It is expressed as R Ｂ１ is -NHCH(C(=O)OH)(CH ２ ) ｍ４１ —O—PO(OH) ２ (wherein m41 is an integer of 1 to 3, preferably 1), —NHCH(C(═O)OH)(CH ２ ) ｍ４２ CH(-O-PO(OH) ２ ) CH ３ (wherein m42 is an integer of 0 to 3, preferably 0), or —NHCH(C(═O)OH)(CH ２ ) ｍ４３ -Ph(-O-PO(OH) ２ ) (wherein m43 is an integer of 1 to 3, preferably 1, and Ph is o-, m-, or p-phenylene, preferably p-phenylene).

[0056] In one embodiment, in the compound represented by formula (1), R Ａ１ is -C(C ｃ１ H ２ｃ１ -O-(C ａ１ H ２ａ１ O) ｂ１１ -) α１ Each c1 is independently 1, each a1 is independently 2, b11 is an integer of 1 to 1,000, preferably an integer of 50 to 1,000, α1 is an integer of 4, and X １ is -(CH ２ ) ｍ１１ -C(=O)-, each m11 independently represents an integer of 1 to 10, preferably 3 to 7, for example, 5, and R Ｂ１ is -NHHC(=O)OH-(CH ２ ) ｍ４１ —O—PO(OH) ２ Each m41 independently represents an integer of 1 to 3, preferably 1.

[0057] Examples of the compound of formula (1) include the following structures: n corresponds to b11 above.

[0058] (Synthesis Method) In one embodiment, the compound represented by formula (1) is synthesized as follows. Note that, although only the case where a polyethylene glycol skeleton is used has been described in this synthesis method, compounds having other skeletons can also be synthesized in the same manner. First, the base moiety R Ａ１ Prepare the compound that will be the raw material for the parent site R. Ａ１ As a compound serving as a raw material for the above, for example, a compound having a group having four carbon atoms directly bonded to polyethylene glycol (TetraPEG) and having the ends capped with a capping agent (e.g., pyrrolidine-2,5-dione-N-yl) can be used. Next, a compound containing a substrate moiety is prepared, and the base moiety R is reacted with the compound in the presence of a Good buffer (e.g., 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid (HEPES)). Ａ１The reaction mixture is added to the starting compound of formula (1), and may be heated if necessary. After the reaction time has elapsed, the mixture is dialyzed to obtain the compound represented by formula (1).

[0059] [Formula (2)] R Ａ２ is a parent moiety, and a commonly used group can be used. Ａ２ The skeleton of Ａ１ In this case, R Ａ１ b11 to b19 are R Ａ２ These are e11 to e19.

[0060] R Ａ２ Each of the repeating units has a repeating unit, and the number of repeating units is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000, and more preferably an integer of 40 to 1,000. The number of repeating units may be an integer of 1 to 800, or may be an integer of 20 to 800.

[0061] R Ａ２ In the formula (I), the polyalkylene glycol backbone is preferably -(C ｄ１ H ２ｄ１ O) ｅ１１ -. d1 is an integer of 1 to 6, preferably 2 to 4, and more preferably 2. In other words, preferably, the polyalkylene glycol skeleton is preferably a polyethylene glycol skeleton. e11 is an integer of 1 to 10,000, preferably an integer of 1 to 1,000, more preferably an integer of 40 to 1,000, for example, an integer of 50 to 1,000. Note that when the right side of the polyalkylene glycol skeleton is a linker X ２ Combine with.

[0062] R Ａ２ In the formula (I), the polyglutamic acid backbone is preferably —(NHCH(C ＊ (=O))CH ２ CH ２ C(=O) ｅ１２ The carbon atom (atom marked with *) of the carbonyl group (C(=O)) of the polyglutamic acid backbone is represented by the linker X ２ It is to be noted that R Ａ２In the whole, at least one * is a linker X ２ The other * may be bonded to an OH group. Ａ２ In the whole, at least one * is a linker X ２ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ２ e12 is an integer of 1 to 10,000, preferably an integer of 1 to 1,000, more preferably an integer of 1 to 800, and particularly preferably an integer of 20 to 800. R Ａ２ may have a polymerization terminating moiety at the end, for example, NH ２ CH(C(=O)OH)CH ２ CH ２ C(=O)-(NHCH(C ＊ (=O))CH ２ CH ２ C(=O) ｂ１２ -NHCH(C(=O)OH)CH ２ CH ２ It may be represented by C(=O)OH.

[0063] R Ａ２ In the formula, the lysine backbone may have the following structure: Ａ２ In the whole, at least one * is a linker X ２ The other * may be bonded to a H atom. Ａ２ In the whole, at least one * is a linker X ２ All * in one unit enclosed in brackets may be bonded to H atoms as long as they are bonded to R. In this case, the H atoms are Ａ２ Included in. e13 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. Ａ２ may have a polymerization terminating moiety at the end, and may be represented by, for example, the following formula: R ｅ３ is not particularly limited, but may be, for example, an OH group, NH ２ group or NH(CH ２ ) ｅ２０ CH ３e20 is an integer of 0 to 10, preferably an integer of 2 to 5.

[0064] R Ａ２ In the formula, the dextran backbone may have the following structure: Ａ２ In the whole, at least one * is a linker X ２ The other * may be bonded to an OH group. Ａ２ In the whole, at least one * is a linker X ２ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ２ Included in. In the formula, e14 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. Ａ２ may have a polymerization terminating moiety at the end, and may be represented by, for example, the following formula:

[0065] R Ａ２ In the above, the alginate backbone may have the following structure: Ａ２ In the whole, at least one * is a linker X ２ The other * may be bonded to an OH group. Ａ２ In the whole, at least one * is a linker X ２ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ２ Included in. In the formula, e15 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000, and e16 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. The sum of e15 and e16 may be an integer of 1 to 10,000, or may be an integer of 1 to 1,000. R Ａ２ may have a polymerization terminating moiety at the end, and may be represented by, for example, the following formula:

[0066] R Ａ２ In the formula, the polyvinyl alcohol backbone is -[C ＊ HCH ２ ]ｅ１７ - may have R Ａ２ In the whole, at least one * is a linker X ２ The other * may be bonded to an OH group. Ａ２ In the whole, at least one * is a linker X ２ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ２ e17 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. Ａ２ may have a polymerization terminating moiety at the end, for example, H—[C ＊ HCH ２ ] ｅ１７ It may be represented by —COOH.

[0067] R Ａ２ In the formula, the polyacrylic acid backbone is -[CH ２ CH(C ＊ (=O)) ｅ１８ - may have R Ａ２ In the whole, at least one * is a linker X ２ The other * may be bonded to an OH group. Ａ２ In the whole, at least one * is a linker X ２ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ２ e18 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. Ａ２ may have a polymerization terminating moiety at the end.

[0068] R Ａ２ In the formula, the polymethacrylic acid backbone is -[CH ２ CCH ３ (C ＊ (=O)) ｅ１９ - may have R Ａ２ In the whole, at least one * is a linker X ２ The other * may be bonded to an OH group. Ａ２In the whole, at least one * is a linker X ２ All * in one unit enclosed in brackets may be bonded to an OH group as long as the OH group is bonded to R Ａ２ e19 is, for example, an integer of 1 to 10,000, preferably an integer of 1 to 1,000. R Ａ２ may have a polymerization terminating moiety at the end.

[0069] When used for a test subject, R Ａ２ is R Ａ１ As with the above, it is preferable that the site has little effect on the subject. For example, R Ａ２ As the backbone, a polyalkylene glycol backbone or a polyglutamic acid backbone can be used.

[0070] R Ａ２ can bind to one or more ligand sites. That is, β1 is an integer of 1 or more, and β2 is an integer of 0 or more. However, the sum of β1 and β2 is not greater than R Ａ２ is the valence of

[0071] β1 is preferably an integer of 2 or more, and β2 is an integer of 0 or more.

[0072] For example, R Ａ２ When is tetravalent, β1 is preferably an integer of 2 to 4, β2 is an integer of 0 to 2, and more preferably β1 is 4.

[0073] For example, when β1 is 4 and β2 is 0, equation (2) is expressed as follows:

[0074] R Ａ２ For example, -C(C ｆ１ H ２ｆ１ -O-(C ｄ１ H ２ｄ１ O) ｅ１１ -) β１ In the formula, f1 is represented by (C ｆ１ H ２ｆ１ ) units, each independently represents an integer of 1 to 6, preferably an integer of 1 to 4, and more preferably 1. ｄ１ H ２ｄ１) unit, e11 is an integer of 1 to 6, preferably an integer of 2 to 4, and more preferably an integer of 2. e11 is an integer of 1 to 1,000, preferably an integer of 1 to 1,000, and more preferably an integer of 50 to 1,000. β1 is an integer of 2 to 4, preferably 3 or 4, and more preferably 4.

[0075] R Ｃ１ is the ligand site. Ｃ１ preferably each independently contain a metal atom. The metal atom is preferably a divalent metal atom, for example, at least one selected from the group consisting of Zn and Mg.

[0076] R Ｃ１ is preferably represented by the following structure: In the formula, * is a bonding moiety, and m2 is an integer of 1 to 6, preferably an integer of 1 to 3, and more preferably 2. M ２＋ is a divalent metal atom.

[0077] R Ｃ２ is a group that does not have a ligand site. Ｃ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms. Ｃ２ is, for example, -(CH ２ ) ｎ３２ -OH, -(CH ２ ) ｎ３２ -COOH, -(CH ２ ) ｎ３２ -H or -(CH ２ ) ｎ３２ -CH ３ Each n32 is independently an integer of 0 to 6.

[0078] X ２ is a linker moiety. ２ is the maternal part R Ａ２ and the ligand site R Ｃ１ or the parent site R Ａ２ and R Ｃ２ and are linked together. X ２ represents a single bond or a divalent or higher organic group. ２ is X １It can have a similar structure.

[0079] X ２ In one embodiment, X is a single bond. ２ In one embodiment, is a divalent or higher organic group.

[0080] X ２ The divalent or higher organic group in the formula (I) contains, for example, at least one selected from the group consisting of alkylene, -C(=O)-, -C(=O)-NH-, -NH-C(=O)-, and phenylene. The hydrogen atom of the divalent or higher organic group may be substituted.

[0081] X ２ are preferably each independently １ Specifically, X ２ is -X ２１ -X ２２ It is represented by -. ２１ R on the side Ａ２ To, X ２２ R on the side Ｃ１ or R Ｃ２ Connect to the respective

[0082] X ２１ has one of the following structures: Ａ２ In ##, X ２２ are connected to the respective single bonds; ＊＊ (CH ２ ) ｍ１１ -C ＃＃ (═O) (x1) [wherein: m11 is each independently an integer of 1 to 10, preferably an integer of 3 to 7, for example, 5]; or Formula (x2): [wherein: m12 is an integer of 1 to 10, preferably 2; m13 is an integer of 1 to 10, preferably 2; and m14 is an integer of 1 to 10, preferably 2.]

[0083] X ２２ has one of the following structures: In the following structures, # represents X ２１ , * is R Ｃ１ or R Ｃ２A single bond; or Formula (z4): [In the formula: x is an integer of 1 to 1,000, preferably an integer of 1 to 800, and more preferably an integer of 1 to 50; y is an integer of 1 to 1,000, preferably an integer of 1 to 800, and more preferably an integer of 1 to 50; the repeating units enclosed in parentheses around x and y may be present in any order in the formula; m18 is an integer of 1 to 6, preferably an integer of 2 to 5, and more preferably 2; and m19 is an integer of 1 to 6, preferably an integer of 1 to 5, and more preferably 1.] (In another example, x may be an integer of 5 to 400, y may be an integer of 5 to 400, and the sum of x and y may be an integer of 5 to 400.)

[0084] Preferably, X ２１ and X ２２ and X are not both single bonds. ２１ is represented by formula (x1) or (x2), and X ２２ is represented by either a single bond or formula (z4).

[0085] -X ２２ -R Ｃ１ Examples of the groups include the following: ２１ Connect to.

[0086] δ1 is a linker X ２ R bonded to Ｃ１ δ1 is an integer of 1 or more. The value of δ1 is the number of linkers X ２ δ1 can vary depending on the structure of the linker X ２ The valence of the atom can be −1.

[0087] R Ａ２ When is a polyalkylene glycol backbone, δ1 can be 1 or more for the entire polyalkylene glycol backbone.

[0088] R Ａ２is a glutamic acid backbone, δ1 can be 1 or greater for the entire glutamic acid backbone. δ1 can be e12 or less for the entire glutamic acid backbone.

[0089] R Ａ２ is a lysine backbone, δ1 can be 1 or greater across the lysine backbone. δ1 can be e13 or less across the lysine backbone.

[0090] R Ａ２ When the dextran backbone is a dextran backbone, δ1 can be 1 or more for the entire dextran backbone. δ1 can be 3×e14 or less for the entire dextran backbone. δ1 can be, for example, e14 or less.

[0091] R Ａ２ is an alginate skeleton, δ1 can be 1 or more for the entire alginate skeleton. δ1 can be 3 × (e15 + e16) or less for the entire alginate skeleton. δ1 may be, for example, (e15 + e16) or less.

[0092] R Ａ２ is a polyvinyl alcohol backbone, δ1 can be 1 or more for the entire polyvinyl alcohol backbone. δ1 can be e17 or less for the entire polyvinyl alcohol backbone.

[0093] R Ａ２ is a polyacrylic acid skeleton, δ1 can be 1 or more for the entire polyacrylic acid skeleton. δ1 can be e18 or less for the entire polyacrylic acid skeleton.

[0094] R Ａ２ When the backbone is a polymethacrylic acid, δ1 can be 1 or more for the entire polymethacrylic acid backbone. δ1 can be e19 or less for the entire polymethacrylic acid backbone.

[0095] δ2 is a linker X ２ R bonded to Ｃ２ δ2 is an integer of 1 or more. The value of δ2 is the number of linkers X ２ The upper limit of δ2 can vary depending on the structure of the linker X ２ The valence of the atom can be −1.

[0096] δ2 can take the same numerical range as δ1.

[0097] In addition, -X ２ (-R Ｃ２ ) δ２ is R in the structure Ｃ１ This refers to a structure that does not contain any of the above.

[0098] In one embodiment, in the compound represented by formula (2), R Ａ２ is -C(C ｆ１ H ２ｆ１ -O-(C ｄ１ H ２ｄ１ O) ｅ１１ -) β１ Each f1 is independently 1, each d1 is independently 2, e11 is an integer of 1 to 10,000, preferably an integer of 1 to 1,000, more preferably an integer of 50 to 1,000, β1 is an integer of 4, and X ２ Ha-(CH ２ ) ｍ１１ -C(=O)-, each m11 independently represents an integer of 1 to 10, preferably 3 to 7, for example, 5, and R Ｃ１ may be represented by the following structure, and the metal atom may be Zn or Mg: ２ and m2 is independently an integer of 1 to 6, preferably an integer of 1 to 3, and more preferably 2.

[0099] Examples of the compound of formula (2) include the following structure: n corresponds to e11 above.

[0100] (Synthesis Method) In one embodiment, the compound represented by formula (2) is synthesized as follows. Note that, although only the case where a polyethylene glycol skeleton is used has been described in this synthesis method, compounds having other skeletons can also be synthesized in the same manner. First, the base moiety R Ａ２ Prepare the compound that will be the raw material for the parent site R. Ａ２As a compound serving as a raw material for the above, for example, a compound having a group having four carbon atoms directly bonded to polyethylene glycol (TetraPEG), in which the terminals are capped with a capping agent (for example, pyrrolidine-2,5-dione-N-yl), can be used. Next, Phostag is prepared, and in the presence of a solvent such as tetrahydrofuran and, if necessary, a pH adjuster such as triethylamine, the parent moiety R Ａ２ The mixture may be heated if necessary. Phostag is a phosphate monoester anion (R-OPO ３ ２－ After the reaction time has elapsed, a solution containing metal ions is added, followed by dialysis to obtain the compound represented by formula (2).

[0101] The hydrogel may have a coordinate bond between the compound represented by formula (1) and the compound represented by formula (2), and may be gelled. For example, when the compound represented by formula (1) is -O-PO(OH), ２ When the compound represented by formula (2) has a metal atom M, the compound represented by formula (2) has a group containing a —O—PO(O － ) ２ and M are thought to coordinate to form the following structure. In the following structure, * indicates bonding to another group. M represents a metal atom.

[0102] The compound represented by formula (1) and the compound represented by formula (2) are preferably mixed in the range of 15 mg / mL:15 mg / mL to 500 mg / mL:500 mg / mL, more preferably in the range of 30 mg / mL:120 mg / mL to 120 mg / mL:30 mg / mL, and particularly preferably in the range of 30 mg / mL:30 mg / mL to 100 mg / mL:100 mg / mL. This configuration allows for stronger coordinate bonding. Furthermore, it becomes possible to control the lifetime (i.e., the time it takes to form a solate).

[0103] The ratio of the number of substrate moieties to the number of ligand moieties is preferably in the range of 1:10 to 10:1, more preferably in the range of 1:2 to 2:1, and even more preferably in the range of 0.8:1 to 1:0.8. This configuration allows for stronger coordination bonding. It also makes it possible to control the lifetime (i.e., the time it takes to form a sol).

[0104] In one aspect, the hydrogel can maintain a gel state for a certain period of time, for example, 1 hour or more, 3 hours or more, 6 hours or more, 24 hours or more, 30 days or more, 60 days or more, or 90 days or more. The gel state can be maintained for, but is not limited to, up to 360 days.

[0105] The hydrogel may further contain a solvent. Examples of the solvent include water, Good's buffer (e.g., 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid (HEPES), and 2-N-morpholinoethanesulfonic acid monohydrate (2-morpholinoethanesulfonic acid monohydrate, MES). In the hydrogel, the total amount of the compound represented by formula (1) and the compound represented by formula (2) may be, for example, in the range of 3 to 50% by mass, specifically, 3 to 10% by mass.

[0106] The hydrogel may contain other compounds or residues thereof, such as compounds used in the synthesis of the compound represented by formula (1) or the compound represented by formula (2) or residues thereof, preservatives, and inorganic ions such as sodium ions and zinc ions.

[0107] The hydrogel of the present invention can encapsulate proteins, drugs, etc., which allows the proteins, drugs, etc. to migrate. Furthermore, degelation of the hydrogel allows the proteins, drugs, etc. to be administered locally.

[0108] In one embodiment, the hydrogel can be formed by mixing a composition containing a compound represented by formula (1) with a composition containing a compound represented by formula (2).

[0109] The compound represented by formula (1) may be contained in the composition in an amount of 50 to 100 mass %, or 80 to 100 mass %.

[0110] The compound represented by formula (2) may be contained in the composition in an amount of 50 to 100 mass %, or 80 to 100 mass %.

[0111] The composition containing the compound represented by formula (1) may contain a fluorescent dye, a protein, etc. in addition to the compound represented by formula (1).

[0112] The composition containing the compound represented by formula (2) may contain, in addition to the compound represented by formula (2), a first enzyme such as a phosphatase, a kinase, etc., which will be described later.

[0113] [Degelation Reaction] When the first enzyme is added to the hydrogel, the coordinate bond between the compound represented by formula (1) and the compound represented by formula (2) is broken, and a degelation reaction (i.e., a sol-forming reaction) can occur. Ｂ１ ga-O-PO(OH) ２ When the first enzyme is a group containing an -OPO group, the first enzyme ３ ２－ The phosphate group acts on the coordinate bond between the group and the metal atom, cleaving the coordinate bond between the compound represented by formula (1) and the compound represented by formula (2), thereby forming a compound in which the phosphate group has been eliminated from the compound represented by formula (1) and a compound represented by formula (2) (dephosphorylation reaction).

[0114] The hydrogel can be used as an injectable hydrogel, whereby the coordination bond is cleaved by a first enzyme in the body, resulting in solization at a predetermined site, i.e., the active ingredient can be released at a targeted site.

[0115] Examples of the first enzyme include phosphatases such as alkaline phosphatase and acid phosphatase, and alkaline phosphatase is preferably used.

[0116] The first enzyme can be added to the hydrogel in an amount of, for example, 0.00001 to 1.0% by mass, specifically, 0.0001 to 0.1% by mass. By adjusting the concentration, the time required for solation can be adjusted. Consequently, sustained release can be achieved during drug administration.

[0117] The first enzyme may be added at, for example, 0.0001 to 10%, specifically 0.001 to 10%, per substrate site. By adjusting the concentration, the time required for solation can be adjusted. Consequently, sustained release can be achieved during drug administration.

[0118] The first enzyme may be added at, for example, 0.0001 to 10%, specifically 0.001 to 10%, per ligand site. By adjusting the concentration, the time required for solation can be adjusted. Consequently, sustained release can be achieved during drug administration.

[0119] When the ratio of substrate sites to ligand sites is in the range of 1:0.9 to 1.1, the first enzyme can be added, for example, at 0.0001 to 10%, specifically 0.001 to 10%, per substrate site. By adjusting the concentration, the time required for solation can be adjusted. Furthermore, sustained release can be achieved during drug administration.

[0120] [Gelation Reaction] A second enzyme may be added to the reaction product (composition after the solation reaction) obtained by the degelation reaction. This causes a coordinate bond between the substrate moiety of the compound represented by formula (1) and the ligand moiety of the compound represented by formula (2), resulting in gelation. For example, R Ｂ１ ga-O-PO(OH) ２ In another example, when the compound represented by formula (1) has a —O—PO(OH) group, the compound represented by formula (2) forms a coordinate bond with the metal atom of the compound represented by formula (1) upon addition of a second enzyme. ２ The compound from which the group has been removed is —O—PO(OH) ２ After the elimination of the group, the compound having a hydroxyl group can form a coordinate bond with the ligand site of the compound represented by formula (2) and form a gel by adding a second enzyme and, if necessary, adenosine triphosphate or the like described below.

[0121] X １ is represented by the same formula as above. Preferably, X １１ is a single bond or any one of formulas (x1) to (x2), and X １２ may be, for example, either formula (z5) or (z6):

[0122] Examples of the second enzyme include kinases such as tyrosine kinase, serine kinase, and threonine kinase, and preferably tyrosine kinase and serine kinase are used.

[0123] The second enzyme may be added in an amount of, for example, 0.0001 to 10%, specifically 0.001 to 10%, per substrate site. By adjusting the concentration, the time required for gelation can be adjusted.

[0124] The second enzyme may be added in an amount of, for example, 0.0001 to 10%, specifically 0.001 to 10%, per ligand site. By adjusting the concentration, the time required for gelation can be adjusted.

[0125] In one embodiment, when the ratio of substrate sites to ligand sites is in the range of 1:0.9 to 1.1, the second enzyme can be added in an amount of 0.0001 to 10%, specifically 0.001 to 10%, per substrate site. By adjusting the concentration, the time required for gelation can be adjusted.

[0126] Along with the second enzyme, adenosine triphosphate (ATP) can be added, resulting in a substrate site where the hydroxyl group is phosphorylated.

[0127] Preferably, the solization reaction and gelation reaction can be repeated, making the product more useful as an injectable hydrogel. The coordination bond is cleaved by a first enzyme in the body, resulting in solization at a predetermined location, and the coordination bond is formed by a second enzyme in the body, resulting in gelation at a predetermined location. In other words, the active ingredient can be released or absorbed at the targeted location.

[0128] <<Second Embodiment>> In the first embodiment, the compound represented by formula (1) and the compound represented by formula (2) were first mixed. However, in the present embodiment, the compound represented by formula (2) and the first enzyme are first mixed, and then the mixture is mixed with the compound represented by formula (1). After mixing, a coordinate bond is formed between the substrate moiety of the compound represented by formula (1) and the ligand moiety of the compound represented by formula (2). This causes the composition to gel. After a certain period of time has passed, the first enzyme reacts with the coordinate bond moiety, causing a sol-forming reaction and cleaving the coordinate bond. The sol-gelation reaction may enable local administration of a drug or the like.

[0129] The certain period of time varies depending on the concentrations of the compound represented by formula (1), the compound represented by formula (2), and the enzyme, and is not particularly limited, and may be, for example, 1 hour, 3 hours, or 18 hours after mixing of the two compositions.

[0130] In this embodiment, the mixture of the compound represented by formula (2) and the first enzyme may be in a solution state. Here, the solution state refers to a liquid state that is not gelled. When the compound represented by formula (1) is added to a composition containing the compound represented by formula (2) and the enzyme, gelation may begin. After a certain period of time has elapsed, the mixture may automatically become a sol.

[0131] The compound represented by formula (1), the compound represented by formula (2), the first enzyme, the second enzyme, other compounds and residues thereof are defined the same as in the first embodiment.

[0132] The composition having the compound represented by formula (1) or the composition having the compound represented by formula (2) and the first enzyme may contain other compounds such as a solvent or residues thereof. The other compounds or residues thereof may be those described above.

[0133] Although the embodiments of the present disclosure have been described above, it will be understood that various changes in form and details can be made without departing from the spirit and scope of the claims.

[0134] The present disclosure will be explained in more detail below through examples, but is not limited to these examples.

[0135] (Synthesis Example 1: Synthesis of TetraPEG-SerPi) TetraPEG-SerPi, a compound represented by formula (1), was synthesized based on the following scheme.

[0136] [Experimental Conditions] <Equipment> Screw vial No. 5 (Maruem, Tokyo, Japan, 0101-07, 20 mL) Spectrapore 7 dialysis membrane (MW = 3,500) (Wako, Osaka, Japan, 536-34441) Freeze dryer (EYELA, Tokyo, Japan, FDU-2200) <Reagents> Pentaerythritol tetra(succinimidylcarboxypentyl)polyoxyethylene (TetraPEG-NHS) (NOF Corporation, Tokyo, Japan, PTE-200HS, Mw: 20,000, n≒100) o-Phospho-L-serine (Wako, Osaka, Japan, 169-21781, Mw: 185.07) HEPES, Nuclease and Protease Test (Nacalai Tesque, Kyoto, Japan, M4R4826, Mw: 238.31) 5 mol / L Sodium Hydroxide Solution (Wako, Osaka, Japan, 196-05375, Mw: 40.00) <Procedure> 12.04 g of HEPES was weighed into a 100 mL beaker and dissolved in 90 mL of water (Milli-Q). 5 mol / L sodium hydroxide solution was added until the pH reached 7.4, and the solution was diluted to 100 mL using a 100 mL measuring cylinder to prepare 500 mM HEPES. Next, 44.42 mg of o-Phospho-L-serine (240 μmol, 48.0 eq.) was added to a screw cap vial No. 5 and dissolved in 4 mL of 500 mM HEPES. Next, 100 mg of TetraPEG-NHS (5.0 μmol, 1.0 eq.) was added and stirred at 37°C for 24 hours. After that, dialysis was carried out against 2 L of Milli-Q using a 3.5 kDa dialysis membrane for 2 days. During dialysis, the solution was changed three times a day. After dialysis, the product was purified by freeze-drying for 3 days. The obtained product was analyzed by nuclear magnetic resonance analysis ( １ The yield was 91%, and the introduction rate of SerPi was 87%.

[0137] (Synthesis Example 2: TetraPEG-PhostagZn ２＋ Synthesis of TetraPEG-PhostagZn, a compound represented by formula (2) ２＋ was synthesized according to the following scheme:

[0138] [Experimental conditions] <Equipment> Screw vial No. 5 (Maruem, Tokyo, Japan, 0101-07, 20 mL) Spectrapore 7 dialysis membrane (MW = 3,500) (Wako, Osaka, Japan, 536-34441) Freeze dryer (EYELA, Tokyo, Japan, FDU-2200) <Reagents> Pentaerythritol tetra(succinimidylcarboxypentyl)polyoxyethylene (TetraPEG-NHS) (NOF Corporation, Tokyo, Japan, PTE-200HS, Mw: 20,000, n≒100) Amino-pendant-Phos-TAG TM Ligand (Amino-pendant Phos-tag TM ligand (Phostag) (Nard Institute, Ltd., Hyogo, Japan, Mw: 540.3), triethylamine (TEA) (TCI, Tokyo, Japan, 121-44-8, Mw: 101.19, SG: 0.73), tetrahydrofuran (with stabilizer) (THF) (Wako, Osaka, Japan, 206-05106, Mw: 72.11), ZnCl ２(Wako, Osaka, Japan, 263-00271, Mw: 136.32) <Procedure> 95.6 mg of Phostag was dissolved in 1.9 mL of THF to prepare a 50 mg / mL Phostag solution. Subsequently, 3.6 mL of THF was placed in a screw tube bottle No. 5, 5.5 μL of TEA (40 μmol, 8.0 eq.) was added, and 432 μL of a 50 mg / mL Phostag solution (40 μmol, 8.0 eq.) was added. 100 mg of TetraPEG-NHS (5.0 μmol, 1.0 eq.) was dissolved therein, and the mixture was stirred at 37°C for 24 hours. The solution was then diluted 5-fold with water (Milli-Q), and 1.3632 g of ZnCl was dialysis-diluted using a 3.5 kDa dialysis membrane. ２ 5 mM ZnCl dissolved in 2 L of Milli-Q ２ The reaction mixture was dialyzed against 2 L of Milli-Q for one day and against 2 L of Milli-Q for one day. The solution was changed three times a day. Some of the reaction mixture was diluted with 5 mM ZnCl ２ The product was not dialyzed against 2 L of Milli-Q but was dialyzed for 2 days against 2 L of Milli-Q. After dialysis, the product was purified by freeze-drying for 3 days. １ The product was analyzed using H-NMR and ultraviolet (UV) measurements. The yield was 82% and the introduction rate was １ The purity was 91% by H-NMR measurement and 109% by UV measurement.

[0139] [Example 1-1] 60 mg / mL of TetraPEG-SerPi was added to HEPES (100 mM, pH 7.35, 25°C) to form a solution. That is, the solution was in a non-gelled state. ２＋ 60 mg / mL was added to HEPES (100 mM, pH 7.35, 25°C) to form a solution. The above two solutions were mixed in equal amounts. After mixing, gelation proceeded rapidly.

[0140] [Examples 1-2 to 1-3] TetraPEG-SerPi and TetraPEG-PhostagZn ２＋ The same procedure as in Example 1-1 was carried out except that the concentrations of were 15 mg / mL and 30 mg / mL, respectively.

[0141] Figure 1 shows the relationship between the time after mixing and the storage modulus (G') and loss modulus (G'') values ​​for Example 1-1. The state of gelation is shown. Immediately after the start of measurement, the storage modulus (G') exceeded the loss modulus (G''), indicating rapid gelation. Figure 2 shows the values ​​of the storage modulus (G') and loss modulus (G''") for Examples 1-2 and 1-3. TetraPEG-SerPi and TetraPEG-PhostagZn ２＋ It was found that as the concentration of α-glucan increases, the storage modulus (G') increases and a stronger gel is formed.

[0142] Example 2: TetraPEG-SerPi and TetraPEG-PhostagZn ２＋ The concentrations of each of these were adjusted to 15 mg / mL and mixed together. Alkaline phosphatase was added to the resulting gel. Specifically, the procedure was as follows.

[0143] [Experimental Conditions] <Reagents> TetraPEG-SerPi (formed in Synthesis Example 1) TetraPEG-PhostagZn ２＋ (Formed in Synthesis Example 2) 100 mM HEPES Alkaline phosphatase from bovine intestinal mucosa (Sigma-Aldrich, St. Louis, USA, P7640) <Procedure> TetraPEG-SerPi and TetraPEG-PhostagZn ２＋ Each of these was added to HEPES to prepare a solution with a concentration of 15 mg / mL. After mixing these two solutions, 1 unit of alkaline phosphatase was added.

[0144] FIG. 3 shows the relationship between the time after mixing and the state of a system to which phosphatase was not added (Example 1-2, 0 units of phosphatase) and a system to which phosphatase was added (Example 2, 1 unit of phosphatase). FIG. 3 was measured at 100 mM HEPES, pH 7.35, and 25°C. As shown in FIG. 3, when alkaline phosphatase was not added, the gel remained even after 18 hours, but by adding alkaline phosphatase, the gel became a sol over time. Specifically, a portion of the gel became a sol after 3 hours, and the gel became completely sol after 18 hours. It is believed that dephosphorylation of the hydrogel was promoted by the phosphatase. This is thought to have caused the network between the two types of TetraPEG to collapse, resulting in the formation of a sol.

[0145] Example 3: TetraPEG-SerPi was added to HEPES to prepare a 15 mg / mL solution. ２＋ was added to HEPES, and 0.1 unit of phosphatase was also added. ２＋ A solution containing 15 mg / mL of the above was prepared. The above two solutions were mixed and then allowed to stand.

[0146] The relationship between the time after mixing and the state of the hydrogel is shown in Figure 4. It was found that the addition of phosphatase caused the formation of a sol over time.

[0147] Example 4 TetraPEG-SerPi (30 mg / mL) was added to 100 μL of a HEPES solution (100 mM, pH 7.4, 25° C.), and 2 μL of a solution containing fluorescein diphosphate (5 mM) was further added to prepare a solution. Separately, TetraPEG-PhostagZn ２＋ (30 mg / mL) was added to 100 μL of HEPES solution (100 mM, pH 7.4, 25° C.) to prepare a solution. When the above two solutions were mixed, gelation proceeded rapidly, and a gel was obtained.

[0148] Then, 800 μL of 100 mM HEPES solution was gently added to the gel and allowed to stand at 37° C. After that, a sample was taken and the concentration of the compound derived from fluorescein diphosphate (leakage rate [%]) was measured. The leakage rate is the molar ratio to the amount added.

[0149] [Concentration measurement] Apparatus: Fluorescence spectrophotometer Conditions: Excitation wavelength 490 nm and fluorescence wavelength 521 nm

[0150] Comparative Example 1 The same procedure as in Example 4 was carried out, except that fluorescein (fluorescein) shown below was used instead of fluorescein diphosphate.

[0151] In Figure 5, the horizontal axis represents elapsed time [days] and the vertical axis represents leakage rate [%]. The results of Example 4 are shown by a solid line, and the results of Comparative Example 1 are shown by a dotted line. As shown in Figure 5, it was found that fluorescein began to elute from the first day in Comparative Example 1. In Example 4, the leakage rate of fluorescein diphosphate, a phosphorylated fluorescent substance, was smaller than the leakage rate of fluorescein in Comparative Example 1. Even after six days, the leakage rate of fluorescein diphosphate was small, indicating that a large amount of fluorescein diphosphate was retained in the gel.

[0152] [Example 5: Adjustment of the amount of phosphatase added] TetraPEG-SerPi (30 mg / mL) was added to 100 μL of HEPE solution (100 mM, pH 7.4, 25°C), and 2 μL of a solution containing 5 mM fluorescein diphosphate was further added to prepare a solution. ２＋ A solution was prepared by adding 30 mg / mL of phosphatase (1 unit) to 100 μL of HEPES solution (100 mM, pH 7.4, 25°C). When the two solutions were mixed, gelation proceeded rapidly, and a gel was obtained.

[0153] Thereafter, 800 μL of 100 mM HEPES solution was gently added onto the gel, and the mixture was allowed to stand at 37° C. Sampling was performed, and the concentration of eluted fluorescein diphosphate (leakage rate [%]) was measured.

[0154] In Figure 6, the horizontal axis represents the elapsed time [days] and the vertical axis represents the leakage rate [%]. The results of Example 5 are indicated by a black circle. For comparison, the results of Example 4 (i.e., the results of an Example that did not contain phosphatase) are used and are represented by a circle in Figure 6. The leakage rate was measured in the same manner as in Example 4. As shown in Figure 6, it was found that fluorescein diphosphate was eluted by adding phosphatase.

[0155] Example 6: Adjustment of the amount of phosphatase added The same procedure as in Example 5 was carried out, except that the amount of phosphatase added was changed to 0.1 unit.

[0156] In Figure 7, the horizontal axis represents the amount of phosphatase added [units], and the vertical axis represents the leakage rate [%]. From the left, Figure 7 shows the leakage rate results after 6 days for Example 4 (addition amount of phosphatase: 0 units), Example 6 (addition amount of phosphatase: 0.1 units), and Example 5 (addition amount of phosphatase: 1 unit). As shown in Figure 7, it was found that the leakage rate could be controlled by the amount of phosphatase added.

[0157] [Example 7: Evaluation of cytotoxicity of gel] [Example 7-1] TetraPEG-SerPi (90 mg / mL) was added to 100 μL of DMEM medium (Dulbecco's modified Eagle's minimum essential medium, phosphate-free, containing 50 mM HEPES (pH 7.4)) to prepare a solution. ２＋ (90 mg / mL) was added to 100 μL of DMEM medium (Dulbecco's modified Eagle's minimum essential medium, phosphate-free, containing 50 mM HEPES (pH 7.4)), and 1.0 × 10 MDA-MB-231 (human breast adenocarcinoma) cells were added. ４The cells / well were added to form a solution. The above two solutions were mixed to form a cell-encapsulating gel. DMEM contains 50 mM HEPES, 10% FBS (fetal bovine serum), and 1% antibiotics. In other words, DMEM does not contain phosphatase. The hydrogel was incubated in 5% CO ２ The cells were incubated for 24 hours at 37°C in the presence of HCl. Afterwards, cytotoxicity was measured using a Live / Dead cell viability kit. The cell viability (ratio of live to dead areas) was 100%.

[0158] [Example 7-2] TetraPEG-SerPi and TetraPEG-Phos-tag-Zn ２＋ The same procedure as in Example 7-2 was carried out except that the concentration of was 120 mg / mL. The cell viability (ratio of live area to dead area) was 95%.

[0159] As shown in Examples 7-1 and 7-2, the hydrogel was shown to be biocompatible.

[0160] [Example 8: Protein leakage] [Example 8-1] TetraPEG-SerPi (30 mg / mL) was added to 100 μL of HEPES solution (100 mM, pH 7.4, 25°C), and 0.5 μM of EGFP (Enhanced Green Fluorescent Protein) was further added to prepare a solution. EGFP (Enhanced Green Fluorescent Protein) is an enhanced green fluorescent protein with EEEEY (phosphorylation reaction substrate sequence) introduced into the C-terminus by gene transfer. EGFP has an arrangement of four glutamic acids and one phosphorylated tyrosine. Separately, Tetra-PEG-Phos-tag-Zn ２＋ (30 mg / mL) was added to 100 μL of HEPES solution (100 mM, pH 7.4, 25° C.) to prepare a solution. When the above two solutions were mixed, gelation proceeded rapidly, and a gel was obtained.

[0161] Thereafter, 800 μL of 100 mM HEPES solution was gently added onto the gel, and the mixture was allowed to stand for 6 days at 37° C. Sampling was performed, and the concentration of the eluted EGFP was measured.

[0162] Example 8-2 The same procedure as in Example 8-1 was repeated, except that 0.5 μM of EGFP-tag (molecular weight 24,000) was used instead of EGFP. EGFP-tag is a protein having a phosphorylated peptide (EEEEpY) at the C-terminus, and is formed by phosphorylating EEEEY introduced at the C-terminus of EGFP.

[0163] [Example 8-3] TetraPEG-SerPi (30 mg / mL) was added to 100 μL of HEPES solution (100 mM, pH 7.4, 25°C), and 0.5 μM of EGFP-tag (molecular weight 24,000) was further added to prepare a solution. ２＋ (30 mg / mL) was added to 100 μL of HEPES solution (100 mM, pH 7.4, 25°C), and 1 unit of phosphatase (ALP) was added to prepare a solution. When the two solutions were mixed, gelation proceeded rapidly, and a gel was obtained. The subsequent procedures were the same as in Example 8-1.

[0164] The conditions for Examples 8-1 to 8-3 are listed below.

[0165] The results are shown in Figure 8. Examples 8-1, 8-2, and 8-3 are shown from left to right in Figure 8. In Example 8-1, it was found that almost all of the EGFP had leaked out after 6 days. In Example 8-2, no EGFP had leaked out, and it can be understood that the EGFP was retained in the gel. In Example 8-3, in which phosphatase was added, 25±8.1% of the protein was released gradually. Note that in Example 8-3, due to the addition of phosphatase, a portion of the gel was degelled. As described above, it was found that the leakage rate of EGFP could be controlled, as shown in Examples 8-2 and 8-3. In particular, it was found that the EGFP carried in the gel could be released gradually by controlling the solubility of the gel, as shown in Example 8-3.

[0166] The hydrogels disclosed herein can be readily gelled or solated, making them suitable for a variety of applications. For example, they can dissolve in response to phosphatases known to be overexpressed on cancer cells, enabling sustained drug release. Furthermore, they can be dissolved by the addition of phosphatases to form hydrogel scaffolds from which three-dimensional tissues can be easily extracted.

Claims

A hydrogel comprising a compound represented by formula (1) and a compound represented by formula (2), wherein at least one substrate site of the compound represented by formula (1) and at least one ligand site of the compound represented by formula (2) are coordinate-bonded. [In formula (1), R Ａ１ is the maternal site; X １ are each independently a single bond or a divalent or higher valent organic group; R Ｂ１ is the substrate site; R Ｂ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms; α1 is an integer of 1 or more; α2 is an integer equal to or greater than 0; The sum of α1 and α2 is R Ａ１ is the valence of; γ1 is an integer equal to or greater than 1; γ2 is an integer equal to or greater than 1; In formula (2), R Ａ２ is the maternal site; X ２ are each independently a single bond or a divalent or higher valent organic group; R Ｃ１ is the ligand site; R Ｃ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms; β1 is an integer of 1 or greater; β2 is an integer equal to or greater than 0; The sum of β1 and β2 is R Ａ２ is the valence of; δ1 is an integer equal to or greater than 1; δ2 is an integer of 1 or more. R Ａ１ and R Ａ２ each independently comprise a polyalkylene glycol backbone, an α-poly-L-glutamic acid backbone, an α-poly-D-glutamic acid backbone, a γ-poly-L-glutamic acid backbone, a γ-poly-D-glutamic acid backbone, an α-poly-L-lysine backbone, an α-poly-D-lysine backbone, a dextran backbone, an alginic acid backbone, a polyvinyl alcohol backbone, a polyacrylic acid backbone, or a polymethacrylic acid backbone. 　 The hydrogel according to claim 2 , wherein the polyalkylene glycol backbone is a polyethylene glycol backbone. R Ａ１ is -C(C ｃ１ H ２ｃ１ -O-(C ａ１ H ２ａ１ O) ｂ１１ -) α１ and c1 is (C ｃ１ H ２ｃ１ ) units are each independently an integer from 1 to 6; a1 is (C ａ１ H ２ａ１ ) units are each independently an integer from 1 to 6; b11 is an integer from 1 to 10,000; α1 is an integer from 2 to 4. The hydrogel according to any one of claims 1 to 3. R Ａ２ is -C(C ｆ１ H ２ｆ１ -O-(C ｄ１ H ２ｄ１ O) ｅ１１ -) β１ and f1 is (C ｆ１ H ２ｆ１ ) units are each independently an integer from 1 to 6; d1 is (C ｄ１ H ２ｄ１ ) units are each independently an integer from 1 to 6; e11 is an integer from 1 to 10,000; β1 is an integer from 2 to 4. The hydrogel according to any one of claims 1 to 4. R Ｂ１ are each independently —O—PO(OH) ２ The hydrogel according to any one of claims 1 to 5, wherein the group is a group containing a group. R Ｂ１ are each independently -O-PO (OH) ２ 、 -NHCH(C(=O)OH)(CH ２ ) ｍ４１ —O—PO(OH) ２ (In the formula, m41 is an integer of 1 to 3). -NHCH(C(=O)OH)(CH ２ ) ｍ４２ -CH(-O-PO(OH) ２ )-CH ３ (wherein m42 is an integer of 0 to 3), or -NHCH(C(=O)OH)(CH ２ ) ｍ４３ -Ph(-O-PO(OH) ２ ) (wherein m43 is an integer of 1 to 3). The hydrogel according to any one of claims 1 to 6. R Ｃ１ The hydrogel according to any one of claims 1 to 7, wherein each independently represents a group containing a metal atom. 　 The hydrogel of claim 8 , wherein the metal atom is a divalent metal atom. R Ｃ１ has the following structure: (wherein * is a bond; m2 is an integer from 1 to 6; M ２＋ is a divalent metal atom. The hydrogel according to any one of claims 1 to 9, having the formula: X １ and X ２ are each independently -(CH ２ ) ｍ１１ -C(=O)-, m11 is an integer from 1 to 10. The hydrogel according to any one of claims 1 to 10. 　 A composition comprising the hydrogel according to any one of claims 1 to 11, and a first enzyme added thereto to decouple at least a portion of the coordinate bonds between the substrate site and the ligand site.

13. The composition of claim 12, wherein the first enzyme is a phosphatase.

14. The composition according to claim 12 or 13, which is gelled by adding a second enzyme.

15. The composition of claim 14, wherein the second enzyme is a kinase. Equation (1): It is represented by; R Ａ１ is the maternal site; X １ are each independently a single bond or a divalent or higher valent organic group; R Ｂ１ is the substrate site; R Ｂ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms; α1 is an integer of 1 or more; α2 is an integer equal to or greater than 0; The sum of α1 and α2 is R Ａ１ is the valence of; γ1 is an integer equal to or greater than 1; A compound wherein γ2 is an integer of 1 or greater. R Ａ１ is a polyethylene glycol backbone (-(C ａ１ H ２ａ１ O) ｂ１１ -); a1 is an integer from 1 to 6; b11 is an integer from 1 to 10,000; X １ is -(CH ２ ) ｍ１１ -C(=O)-; m11 is an integer from 1 to 10; R Ｂ１ are each independently —NHCH(C(═O)OH)—(CH ２ ) ｍ１ —O—PO(OH) ２ and m1 is an integer from 1 to 3; α1 is 4; α2 is 0, 17. The compound of claim 16. Equation (2): It is represented by; R Ａ２ is the maternal site; X ２ are each independently a single bond or a divalent or higher valent organic group; R Ｃ１ is the ligand site; R Ｃ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms; β1 is an integer of 1 or greater; β2 is an integer equal to or greater than 0; The sum of β1 and β2 is R Ａ２ is the valence of; δ1 is an integer equal to or greater than 1; A compound wherein δ2 is an integer of 1 or greater. R Ａ２ is a polyethylene glycol backbone (-(C ｄ１ H ２ｄ１ O) ｅ１１ -); d1 is an integer from 1 to 6; e11 is an integer from 1 to 10,000; X ２ is -(CH ２ ) ｍ１１ -C(=O)-; m11 is an integer from 1 to 10; R Ｃ１ each independently have the following structure: having * is a binding moiety; m2 is an integer from 1 to 6; 19. The compound of claim 18. 　 A compound of claim 18 or 19; A first enzyme; A composition comprising: 　 The composition of claim 20 further comprising a compound of formula (1): A composition comprising a compound represented by the formula: [In the formula: R Ａ１ is the maternal site; X １ are each independently a single bond or a divalent or higher valent organic group; R Ｂ１ is the substrate site; R Ｂ２ are each independently a hydroxyl group, a carboxyl group, a hydrogen atom, or an alkyl group having 1 to 6 carbon atoms; α1 is an integer of 1 or more; α2 is an integer equal to or greater than 0; The sum of α1 and α2 is R Ａ１ is the valence of; γ1 is an integer equal to or greater than 1; γ2 is an integer of 1 or greater.

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