Compound, hydrogel, and uses thereof

Abstract

To provide a hydrogel exhibiting thixotropy.SOLUTION: The invention provides a compound represented by a predetermined general formula (1) or general formula (2). In the formula (1), X is represented by a predetermined general formula (3); in the formula (2), X and Y are each independently represented by a predetermined general formula (3); and in the formula (3), R1, R2 and R3 each independently represent a hydrogen atom, a halogen atom, or an alkyl group having 1 to 4 carbon atoms.SELECTED DRAWING: Figure 3

Description

Technical Field

[0001] The present disclosure relates to hydrogels.

Background Art

[0002] In recent years, supramolecular hydrogels have attracted attention as medical materials. Non-Patent Document 1 discloses that an amino sugar derivative in which an NPmoc group (p-nitrobenzyloxycarbonyl) is introduced into an amino sugar self-assembles in water to form a supramolecular hydrogel.

Prior Art Documents

Non-Patent Documents

[0003]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] The supramolecular hydrogel described in Non-Patent Document 1 shows a gel-sol transition in response to a reducing stimulus such as Na2S2O4, but does not have thixotropy. However, for example, as an injectable hydrogel, it is desired to have thixotropy. Therefore, a technique capable of providing a hydrogel having thixotropy is required.

Means for Solving the Problems

[0005] The present disclosure can be realized in the following forms.

[0006] (1) According to one aspect of the present disclosure, the following general formula (1) or the following general formula (2):

Chemical Formula

[0007] (2) In the compound according to (1) above, in the general formula (1), X may be in the para position with respect to the bonding position with the amino sugar. According to the compound of this form, a hydrogel with more excellent thixotropy can be provided.

[0008] (3) In the compound according to (1) or (2) above, in the general formula (3), R 1 , R 2 , and R 3 may be a hydrogen atom or a halogen atom. According to the compound of this form, a hydrogel with even more excellent thixotropy can be provided.

[0009] (4) According to another form of the present disclosure, a hydrogel containing the compound according to any one of (1) to (3) above and a solvent containing water is provided. According to the hydrogel of this form, it has thixotropy.

[0010] (5) In the hydrogel according to (4) above, the compound may be contained in an amount of 0.7% by mass or more and 10% by mass or less. According to the hydrogel of this form, it has more excellent thixotropy.

[0011] (6) According to another aspect of the present disclosure, a pharmaceutical composition containing the hydrogel described in the above (4) or (5) is provided. According to the pharmaceutical composition of this aspect, a pharmaceutical composition containing a hydrogel having thixotropy can be provided.

[0012] (7) According to another aspect of the present disclosure, the use of the compound described in any one of the above (1) to (3) for the production of a hydrogel is provided. According to this aspect, a hydrogel having thixotropy can be provided.

[0013] It should be noted that the present disclosure can be realized in various forms, for example, in the form of a method for producing a hydrogel, a composition for producing a hydrogel, a kit for producing a hydrogel, and the like.

Brief Description of the Drawings

[0014]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Mode for Carrying Out the Invention

[0015] According to one embodiment of the present disclosure, a compound represented by the following general formula (1) or the following general formula (2) is provided. In formula (1), X is represented by the following general formula (3). In formula (2), X and Y are each independently represented by the following general formula (3). In formula (3), R 1 , R 2 , and R 3 each independently represent a hydrogen atom, a halogen atom, or an alkyl group having 1 to 4 carbon atoms.

[0016]

Chemical formula

[0017]

Chemical formula

[0018]

Chemical formula

[0019] In the above general formula (1) or the above general formula (2), the bonding position of X is not particularly limited and may be located at the ortho position, meta position, or para position with respect to the bonding position to the amino sugar. However, from the viewpoint of improving thixotropy, it is preferably located at the ortho position or para position, and more preferably located at the para position. In the above general formula (2), the bonding position of Y is not particularly limited and may be located at the ortho position, meta position, or para position with respect to the bonding position of X. However, from the viewpoint of improving thixotropy, it is preferably located at the ortho position or meta position. In the above general formula (2), X and Y may be the same or different from each other, but from the viewpoint of suppressing the complication of compound synthesis, they are preferably the same.

[0020] In the above general formula (3), the "alkyl group having 1 to 4 carbon atoms" may be linear or branched, but from the viewpoint of making the thixotropic property better, it is preferably linear. The alkyl group having 1 to 4 carbon atoms is not particularly limited, and examples thereof include a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, and the like. From the viewpoint of making the thixotropic property better, the alkyl group having 1 to 4 carbon atoms preferably has 1 to 3 carbon atoms, more preferably 1 to 2 carbon atoms, and even more preferably 1 carbon atom.

[0021] The halogen atom in the above general formula (3) is not particularly limited, and examples thereof include a fluorine atom (F), a chlorine atom (Cl), a bromine atom (Br), an iodine atom (I), etc. From the viewpoint of making the thixotropic property better, it is preferably a fluorine atom or a chlorine atom, and more preferably a fluorine atom.

[0022] In the above general formula (3), R 1 , R 2 , and R 3 may be the same or different from each other, but from the viewpoint of suppressing the complication of the synthesis of the compound, it is preferably the same. In the above general formula (3), R 1 , R 2 , and R 3 are preferably a hydrogen atom or a halogen atom, respectively, from the viewpoint of making the thixotropic property better. The number of alkyl groups contained in the above general formula (3) is preferably 2 or less, and more preferably 1 or less, from the viewpoint of making the thixotropic property better.

[0023] The compound represented by the above general formula (1) or the above general formula (2) is preferably represented by the above general formula (1) from the viewpoint of making the thixotropic property better. As the compound represented by the above general formula (1), X is located at the ortho position or the para position with respect to the bonding position with the amino sugar, and R 1, R 2 , and R 3 is preferably a hydrogen atom or a halogen atom, respectively, X is located at the para position with respect to the bonding position with the amino sugar, and R 1 , R 2 , and R 3 is more preferably a hydrogen atom or a halogen atom, respectively. As the compound represented by the general formula (1), a compound represented by the following chemical formula (4) (hereinafter, also referred to as "GlcN-2-CH3Pmoc"), a compound represented by the following chemical formula (5) (hereinafter, also referred to as "GlcN-4-CH3Pmoc"), or a compound represented by the following chemical formula (6) (hereinafter, also referred to as "GlcN-4-CF3Pmoc") is even more preferable, and a compound represented by the following chemical formula (5) or a compound represented by the following chemical formula (6) is particularly even more preferable.

[0024]

Chemical formula

[0025]

Chemical formula

[0026]

Chemical formula

[0027] The compound represented by the general formula (1) or the general formula (2) is a glucosamine derivative and can be synthesized, for example, by the following method. More specifically, first, a precursor compound is synthesized by the following reaction steps according to the methods described in Gonzalez, Maria Isabel et al., WO2002040008 A2, 2002, Dominianni, Samuel J. et al., WO98000403 A1, 1998, etc.

[0028]

Chemical formula

[0029] Next, as shown in the following reaction steps, GlcNH2·HCl is stirred in advance under ice-cooling together with DIEA (N,N-diisopropylethylamine) and DMF (N,N-dimethylformamide), and then after returning to room temperature, the above-mentioned precursor compound is added under ice-cooling, and then gradually returned to room temperature and stirred, whereby it can be synthesized. In the present disclosure, "room temperature" generally means about 10°C to about 35°C.

[0030]

Chemical formula

[0031] The compound represented by the above general formula (1) or the above general formula (2) can be produced by relatively short-step organic chemical synthesis, so that it is possible to suppress the complication of the compound production process. Further, since the compound represented by the above general formula (1) or the above general formula (2) is a glucosamine derivative synthesized using glucosamine, which is a kind of amino sugar, as one of the raw materials, the raw materials can be obtained relatively easily, and as a result, it is possible to suppress an increase in the cost required for the production of the compound.

[0032] According to another aspect of the present disclosure, there is provided a hydrogel containing the compound represented by the above general formula (1) or the above general formula (2) and a solvent containing water. As shown in the examples described later, an aqueous solution containing the compound of the present disclosure exhibits thixotropy.

[0033] In the present disclosure, "thixotropy" means the property that a hydrogel dissolves and turns into a liquid (sol) in response to stress and gels again when the stress is released. Whether or not a hydrogel has thixotropy can be evaluated using a rheometer. More specifically, a container (inner diameter: about 2 cm) containing the hydrogel is placed on the stage of the rheometer, and a 20-mm stainless steel parallel plate is brought into contact at a GAP of 1500 μm for vibration measurement. Vibration measurement (1.0 rad / s) is performed at a strain of 0.1%, and after measuring the time changes of the storage modulus G' and the loss modulus G'', vibration measurement (1.0 rad / s) is performed at a strain of 150% for the purpose of sol-gel transition by stress. Then, vibration measurement (1.0 rad / s) is performed again at a strain of 0.1%, and this strain change (from 0.1% to 150%) is repeated three times. When, at a strain of 0.1%, the value of G' > G'' indicating the behavior of the gel state is shown, at a strain of 150%, the value of G' < G'' indicating the behavior of the sol state is shown, and furthermore, when vibration measurement is performed with the strain returned to 0.1% and the stress suppressed, and the value of G' > G'' indicating the behavior of the gel state again is shown, it can be evaluated that the hydrogel has thixotropy.

[0034] Also, when the hydrogel prepared in a glass screw tube bottle is shaken vigorously to apply mechanical stress to break down the gel, and then left standing at room temperature to observe the state change, the shorter the time taken to gel again, the higher the thixotropy can be evaluated.

[0035] The mechanism by which the compound of the present disclosure forms a hydrogel and has thixotropy is not clear. However, the following putative mechanism is presumed.

[0036] FIG. 1 is a schematic diagram showing a presumed mechanism by which the compound of the present disclosure forms a hydrogel and has thixotropy. In order for the compound to form a hydrogel, it is presumed that the compound aggregates in an aqueous solution to form a fibrous structure, and further that the fibrous structure needs to be networked. Furthermore, in order for thixotropy to be exhibited, it is presumed that the network of the fibrous structure collapses once by mechanical stress and then is restored and networked again when the stress is released. The compound represented by the above general formula (1) or the above general formula (2) can self-assemble in an aqueous solution, and since the fibrous structure is networked, it is considered that a hydrogel can be formed. In addition, it is considered to have thixotropy because the network of the fibrous structure is networked again even after being collapsed by mechanical stress.

[0037] In the hydrogel of the present disclosure, the content of the compound represented by the above general formula (1) and / or the above general formula (2) is not particularly limited, but from the viewpoint of better thixotropy, it is preferably 0.7% by mass or more based on the total mass of the hydrogel, and from the viewpoint of solubility, it is preferably 10% by mass or less. The hydrogel of the present disclosure preferably contains the compound represented by the above general formula (1) or the above general formula (2) in an amount of 0.7% by mass or more and 10% by mass or less, more preferably 1.0% by mass or more and 8% by mass or less, and even more preferably 1.5% by mass or more and 5% by mass or less.

[0038] The hydrogel of the present disclosure may contain only one kind of the compound represented by the above general formula (1) and / or the above general formula (2), or may contain two or more kinds. From the viewpoint of better thixotropy, the hydrogel of the present disclosure preferably contains one or more kinds of the compound represented by the above general formula (1).

[0039] The solvent containing water is not particularly limited, and examples thereof include water, physiological saline, buffer solutions, etc. The buffer solution is not particularly limited, and examples thereof include HEPES-NaOH buffer, MES-NaOH buffer, phosphate buffer, etc. In addition to containing water, the solvent containing water may be miscible with water and contain other solvents in which the compound is soluble. The other solvent is not particularly limited, and examples thereof include DMSO (Dimethyl sulfoxide), DMF (N,N-dimethylformamide), alcohol, etc. The alcohol is not particularly limited, and examples thereof include ethanol, hexafluoroisopropanol, etc. From the viewpoint of better thixotropy, the solvent containing water is preferably a buffer solution containing DMSO.

[0040] The hydrogel of the present disclosure may contain other components within a range where thixotropy is not impaired. The other components are not particularly limited, and examples thereof include emulsifiers, suspending agents, surfactants, stabilizers, flavoring agents, fragrances, excipients, preservatives, binders, diluents, isotonic agents, soothing agents, bulking agents, disintegrants, buffers, coating agents, lubricants, coloring agents, sweeteners, thickeners, flavoring and odor-correcting agents, solubilizing aids, etc.

[0041] The aqueous solution containing the compound of the present disclosure has thixotropy. Therefore, the hydrogel of the present disclosure can be used as a pharmaceutical composition encapsulating a desired substance. Although this substance is not particularly limited, examples include biopharmaceuticals such as drugs, proteins, and nucleic acids. That is, the hydrogel of the present disclosure can be used as, for example, a local delivery carrier or a sustained-release delivery carrier for biopharmaceuticals such as drugs, proteins, and nucleic acids. Although the drug is not particularly limited, examples include anticancer agents, anti-inflammatory agents, antithrombotic agents, antibiotics, antiviral agents, and the like. The method for encapsulating substances such as drugs, proteins, and nucleic acids is not particularly limited, and the substance may be mixed when preparing the hydrogel, or the substance may be mixed after preparing the hydrogel. From the viewpoint of dispersibility, it is preferable to mix the substance when preparing the hydrogel. Since the hydrogel of the present disclosure has thixotropy, it can be used as an injectable hydrogel. In addition, the hydrogel of the present disclosure may be used for cell encapsulation, three-dimensional cell culture, organoids, biomaterials, cell transplantation, cell therapy, tissue modification, tissue regeneration, tissue transplantation, regenerative medicine, 3D printing, wound dressing materials, wound treatment, in vitro models for research or testing of diagnostic agents or therapeutic agents, and the like. According to the hydrogel of the present disclosure, since the molecular weight of the compound is relatively small, long-term residues that may lead to toxicity in the living body can be suppressed.

[0042] The hydrogels of the present disclosure can be prepared, for example, by the following methods. Heating may be performed during the preparation, or it may be prepared without heating. By preparing without heating, when various drugs, proteins, etc. are embedded, it is possible to suppress the denaturation of the drugs and proteins. As a method for producing a hydrogel when heating is performed, first, the compound represented by the above general formula (1) and / or the above general formula (2) is mixed with a solvent containing water. Then, it is mixed under heating. When mixing, ultrasonic waves may be irradiated as necessary. Then, it is allowed to stand at room temperature. As a method for producing a hydrogel without heating, first, the compound represented by the above general formula (1) and / or the above general formula (2) is mixed with a solvent containing water. Then, it is mixed without heating. When mixing, ultrasonic waves may be irradiated as necessary. Then, it is allowed to stand at room temperature.

[0043] Examples of more specific production methods will be described below. First, the compound represented by the above general formula (1) and / or the above general formula (2) is dissolved in a solvent such as DMSO, and then a solution is prepared by mixing a solvent containing water. When preparing by heating, the prepared solution is irradiated with ultrasonic waves, heated with a heat gun, etc., and then a hydrogel is obtained by allowing it to stand at room temperature. When preparing without heating, the prepared solution is irradiated with ultrasonic waves, well mixed, and then a hydrogel is obtained by allowing the resulting dispersion to stand at room temperature.

[0044] According to other aspects of the present disclosure, a pharmaceutical composition comprising the above-described hydrogel is provided. The pharmaceutical composition may contain a pharmacologically acceptable carrier, medium, additive, etc., within a range where the thixotropy of the hydrogel is not impaired. The carrier is not particularly limited, and examples thereof include liquid carriers such as glycerin, peanut oil, polyvinylpyrrolidone, olive oil, ethanol, benzyl alcohol, and propylene glycol. The medium is not particularly limited, and examples thereof include sterilized water, physiological saline, vegetable oil, solvents, etc. The additive is not particularly limited, and examples thereof include emulsifiers, suspending agents, surfactants, stabilizers, flavoring agents, fragrances, excipients, preservatives, binders, diluents, isotonic agents, soothing agents, bulking agents, disintegrants, buffers, coating agents, lubricants, coloring agents, sweeteners, thickeners, flavoring and odor-correcting agents, solubilizing aids, etc.

Example

[0045] Hereinafter, the present invention will be described more specifically by way of examples, but the present invention is not limited to the following examples.

[0046] 1. Synthesis of Compound Compounds (GlcN-2-CH3Pmoc, GlcN-4-CH3Pmoc, GlcN-4-CF3Pmoc) were synthesized and used in the experiments described below. In the following description, these three types of compounds are also collectively referred to as "GlcN-XPmoc". The precursor compounds (2-CH3Pmoc-ONp ((2-Methylphenyl)methyl 4-nitrophenyl carbonate), 4-CH3Pmoc-ONp ((4-Methylphenyl)methyl 4-nitrophenyl carbonate: Gonzalez, Maria Isabel et al., WO2002040008 A2, 2002), 4-CF3Pmoc-ONp (4-Nitrophenyl [4-(trifluoromethyl)phenyl]methyl carbonate: Dominianni, Samuel J. et al., WO9800403 A1, 1998)) were synthesized according to the previously reported methods. GlcN-XPmoc was synthesized according to the following synthetic scheme.

[0047]

Chemical formula

[0048] Details regarding the synthesis of GlcN-2-CH3Pmoc are described below.

[0049]

Chemical formula

[0050] Heat at 120 °C and add dry DMF (5.0 mL), GlcNH2·HCl (259 mg, 1.2 mmol), and DIEA (210 μL, 1.2 mmol) to a 30 mL eggplant flask purged with Ar. Start stirring under ice-cooling. Then, return to room temperature. After 1 hour, add 2-CH3Pmoc-ONp (287 mg, 1.0 mmol) under ice-cooling. Then, gradually return to room temperature and stir for 14 hours. Monitor the reaction by TLC. After confirming the appearance of a new spot and the disappearance of the starting material, add toluene (2 mL), and then distill off the solvent under reduced pressure using a high-boiling-point evaporator. Then, wash with dH2O (2.0 mL × 4) and DCM (2.0 mL × 4), filter by suction using a Kiriyama funnel, collect the residue, and dry it under vacuum in a desiccator. Further, wash with dH2O (1.5 mL × 3), MeOH (1.0 mL), and DCM (1.5 mL × 3), and then dry the collected residue under vacuum in a desiccator. Thus, GlcN-2-CH3Pmoc (225 mg, 0.69 mmol, 69%) was obtained as a white solid. 1 1H NMR (DMSO-d6, + D2O, 400 MHz, rt): δ (ppm) = 7.28 - 7.33 (m, ArH, 1H), 7.14 - 7.23 (m, ArH, 3H), 6.90 (d, J H = 8.2 Hz, NH, β-anomer), 6.40 - 6.45 (m, NH, α-anomer), 5.00 (s, 2H), 4.95 (d, J H = 3.2 Hz, 0.73H, 1 CH,α-anomer), 4.38 (d, J H = 8.2 Hz, 0.27H, 1 CH, β-anomer), 3.03 - 3.57 (m, 6H, 2,3,4,5,6 CH), 2.28 (s, 3H). 1313C NMR (DMSO-d6, + D2O, 600 MHz, rt): δ (ppm) = 18.61, 56.36, 56.45, 61.13, 63.90, 70.30, 71.10, 72.13, 90.78, 125.96, 128.16, 128.62, 130.18, 135.17, 136.50, 156.36. HRMS (ESI, positive) [M (C 15 H 21 NO7) + Na] + : m / z = 350.1216; Found: 350.1258.

[0051] Details regarding the synthesis of GlcN-4-CH3Pmoc are described below.

[0052]

Chemical Structure

[0053] Heated at 120 °C, dry DMF (5.0 mL), GlcNH2·HCl (130 mg, 0.60 mmol), and DIEA (0.10 mL, 0.60 mmol) were added to a 30 mL eggplant flask with Ar substitution, and stirring was started under ice-cooling. Then, the mixture was returned to room temperature, and after 1 hour, 4-CH3Pmoc-ONp (136 mg, 0.47 mmol) was added under ice-cooling. Then, the mixture was gradually returned to room temperature and stirred for 23 hours. The reaction was monitored by TLC. After confirming the appearance of a new spot and the disappearance of the starting material, toluene (2 mL) was added, and then the solvent was distilled off under reduced pressure using a high-boiling-point evaporator. Then, it was washed with dH2O (2.0 mL × 4) and DCM (2.0 mL × 4), and then suction filtered through a Kiriyama funnel to collect the residue, which was then dried in vacuo in a desiccator. Furthermore, it was washed with dH2O (1.5 mL × 3), MeOH (1.0 mL), and DCM (1.5 mL × 3), and then the collected residue was dried in vacuo in a desiccator. Thus, GlcN-4-CH3Pmoc (98 mg, 0.30 mmol, 64%) was obtained as a white solid. 11H NMR (DMSO-d6, + D2O, 400 MHz, rt): δ (ppm) = 7.17 - 7.21 (m, ArH, 2H), 7.10 - 7.13 (m, ArH, 2H), 6.97 - 7.02 (d, J H = 8.2 Hz, NH, β-anomer), 6.79 (m, NH, α-anomer), 4.88 - 5.00 (br.s, 2.7H, + 1 CH, α-anomer), 4.38 (d, J H = 8.1 Hz, 0.32H, 1 CH, β-anomer), 2.96 - 3.54 (m, 6H, 2,3,4,5,6 CH), 2.23 (s, 3H). 13 13C NMR (DMSO-d6, + D2O, 600 MHz, rt): δ (ppm) = 21.04, 56.54, 61.23, 61.34, 65.57, 70.42, 70.99, 71.06, 72.21, 76.83, 90.90, 95.62, 128.26, 129.21, 129.23, 129.31, 134.27, 137.45, 156.56, 156.67. HRMS (ESI, positive) [M (C 15 H 21 NO7) + Na] + : m / z = 350.1216; Found: 350.1197.

[0054] Details regarding the synthesis of GlcN-4-CF3Pmoc are described below.

[0055]

Chemical Structure

[0056] Heated at 120 °C, dry DMF (5.0 mL), GlcNH2·HCl (311 mg, 1.2 mmol), and DIEA (0.44 mL, 2.5 mmol) were added to a 30 mL eggplant flask with Ar substitution, and stirring was started under ice cooling. Then, it was returned to room temperature, and after 1 hour, CF3Pmoc-ONp (342 mg, 1.1 mmol) was added under ice cooling. Then, it was gradually returned to room temperature and stirred for 23 hours. After confirming the appearance of a new spot and the disappearance of the raw material by TLC, toluene (2 mL) was added, and then the solvent was distilled off under reduced pressure using an evaporator for high-boiling solvents. Then, it was washed with dH2O (2.0 mL × 2) and DCM (2.0 mL × 2), then suction filtered through a Kiriyama funnel, and the residue was collected and vacuum dried in a desiccator. Furthermore, it was washed with dH2O (2.0 mL × 2), MeOH (1.0 mL), and DCM (2.0 mL × 2), and then the recovered residue was vacuum dried in a desiccator. Thus, GlcN-4-CF3Pmoc (210 mg, 0.55 mmol, 50%) was obtained as a white solid. 1 1H NMR (DMSO-d6, + D2O, 400 MHz, rt): δ (ppm) = 7.73 (d, J H = 8.2 Hz, ArH, 2H), 7.58 (d, J H = 8.2, ArH, 2H), 7.05 (d, J H = 8.2 Hz, NH, β-anomer), 6.46 (d, J H = 4.6 Hz, NH,α-anomer), 5.11 (s, 2H), 4.95 (d, J H = 3.2 Hz, 1 CH,α-anomer, 0.71H), 4.41 (d, J H = 8.2 Hz, 1 CH, β-anomer, 0.29H), 3.02-3.72 (m, 6 H, 2,3,4,5,6 CH). 1313C NMR (DMSO-d6, + D2O, 600 MHz, rt): δ (ppm) = 56.52, 56.61, 61.22, 61.33, 64.52, 64.72, 70.45, 70.47, 71.07, 72.23, 76.86, 90.90, 125.45, 125.52, 125.54, 125.57, 128.24, 128.29, 128.30, 128.44, 128.65, 142.36, 156.28, 156.34. HRMS (ESI, positive) [M (C 15 H 18 F3NO7) + Na] + : m / z = 404.0933; Found: 404.0954.

[0057] As a compound of the comparative example, GlcN-NPmoc was synthesized according to the following synthetic scheme.

[0058]

Chemical formula

[0059] NaHCO3 (39 mg, 0.46 mmol) was added to H2O (1.0 mL) containing GlcNH2·HCl (50 mg, 0.23 mmol). The mixture was cooled in an ice bath, and then a solution of NPmoc-Cl (60 mg, 0.28 mmol) in 1,4-dioxane (0.50 mL) was added dropwise. Further, 1,4-dioxane (0.83 mL) and H2O (0.50 mL) were added, and the resulting mixture was stirred at room temperature for 1.5 hours. Then, the solvent was distilled off under reduced pressure using an evaporator, and CH2Cl2 (5 mL) was added to the obtained residue. The formed precipitate was washed with CH2Cl2 (15 mL) and H2O (5 mL), and then dried in vacuo in a desiccator. Thereby, GlcN-NPmoc (61 mg, 74%) was obtained as a white solid. 1 1H NMR (DMSO-d6 + D2O, 400 MHz, rt): δ (ppm) = 8.24 (d, J H= 8.0 Hz, ArH, 2H), 7.63 (d, J H = 8.8 Hz, ArH, 2H), 7.14 (d, J H = 8.0 Hz, slightly remained even after the addition of D2O, NH, β-anomer), 6.50 (d, J H = 4.8 Hz, slightly remained even after the addition of D2O, NH, α-anomer), 5.15 (s, 2H), 4.96 (d, J H = 3.2 Hz, 0.67H, 1 CH, α-anomer), 4.42 (d, J H = 8.0 Hz, 0.32H, 1 CH, β-anomer), 3.67 - 3.03 (m, 6H, 2,3,4,5,6 CH), Duplicated signals distinguishable was assigned to the two anomeric forms. 13 C NMR (DMSO-d6, 100 MHz, rt): δ (ppm) = 56.67, 59.06, 61.28, 61.34, 64.22, 70.51, 71.07, 71.23, 72.27, 74.39, 77.03, 90.87, 95.56, 123.74, 128.34, 145.51, 147.10, 156.04. HRMS (ESI, positive): Calcd. for [M(C 14 H 18 N2O9) + Na] + : m / z = 381.0910; Found: 381.0894.

[0060] 2. Method for preparing hydrogel Using the synthesized GlcN-XPmoc, a hydrogel was prepared. For the preparation of the hydrogel, as shown below, a method with heating and a method without heating were used.

[0061] (1) Method with heating Weighed GlcN-XPmoc (4.0 mg) was dissolved in 20 μL of DMSO to prepare a 200 mg / mL GlcN-XPmoc solution. 10 μL of the prepared solution was placed in a glass screw vial (No. 02, 11.7×35 mm, manufactured by Maruemu), mixed with 190 μL of 100 mM HEPES-NaOH buffer (pH 7.4), irradiated with ultrasonic waves, and heated with a heat gun to adjust the solution. The obtained solution was allowed to stand at room temperature to obtain a hydrogel.

[0062] (2) Method without heating Weighed GlcN-XPmoc (4.0 mg) was dissolved in 20 μL of DMSO to prepare a 200 mg / mL GlcN-XPmoc solution. 10 μL of the prepared solution was placed in a glass screw vial (No. 02, 11.7×35 mm, manufactured by Maruemu), mixed with 190 μL of 100 mM HEPES-NaOH buffer (pH 7.4), irradiated with ultrasonic waves, and thoroughly mixed. The obtained dispersion was allowed to stand at room temperature to obtain a hydrogel.

[0063] 3. Evaluation of hydrogel-forming ability The concentration dependence regarding the formation of the hydrogel was investigated. As the compounds, GlcN-2-CH3Pmoc, GlcN-4-CH3Pmoc, GlcN-4-CF3Pmoc, and GlcN-NPmoc were used. For the preparation of the hydrogel, the preparation method with heating was used. As the aqueous solution, 100 mM HEPES-NaOH buffer (pH 7.4) containing 5.0% by volume of DMSO was used. The concentrations of the compounds were 0.25% by volume, 0.50% by volume, 0.75% by volume, and 1.0% by volume. The hydrogel-forming ability was evaluated by inverting the sample vial and visually observing whether it flowed down.

[0064] Figure 2 is an explanatory diagram showing the evaluation results of the ability to form a hydrogel. In Figure 2 and the figures described later, "Gel" indicates a gel state, "pGel" indicates a partially gel state, "Ppt" indicates a precipitation state, and "Sol" indicates a solution state. Both "Ppt" and "Sol" correspond to non-gel states. In addition, in Figure 2, the molar concentrations of each compound are shown together. When the above-described hydrogel preparation method was used, a hydrogel was formed at a concentration of about 0.75% by mass or more, including a partially gel state. In addition, for GlcN-2-CH3Pmoc, both a gel state and a precipitation state were observed at 1.0% by volume.

[0065] 4. Evaluation of Thixotropy of Hydrogel by Visual Observation Regarding the hydrogel, the thixotropy was evaluated by visual observation. As the compounds, GlcN-4-CH3Pmoc, GlcN-4-CF3Pmoc, and GlcN-NPmoc were used. For the preparation of the hydrogel, a preparation method with heating was used. As the aqueous solution, 100 mM HEPES-NaOH buffer (pH 7.4) containing 5.0% by volume of DMSO was used. The concentration of the compound was 1.0% by volume. Mechanical stress was applied to disintegrate the gel by strongly shaking the hydrogel prepared in a glass screw tube (No. 02, 11.7 × 35 mm, manufactured by Maruemu). Then, the state change was visually observed by allowing it to stand at room temperature to simply evaluate the thixotropy. In visual observation, when gelation was observed again from the sol state after stress release, it was evaluated as having thixotropy, and when no re-gelation was observed, it was evaluated as having no thixotropy. In addition, it can be evaluated that the shorter the time required for re-gelation after stress release, the higher the thixotropy.

[0066] Figure 3 is an explanatory diagram showing the evaluation results of thixotropy by visual observation. For GlcN-4-CH3Pmoc and GlcN-4-CF3Pmoc, they showed the behavior of gelling again within 5 minutes after stress release, indicating good thixotropy. In contrast, for GlcN-NPmoc, after being liquefied by stress, no re-gelling was confirmed even 30 minutes after stress release, indicating that it does not have thixotropy or has extremely low thixotropy.

[0067] 5. Quantitative evaluation of the thixotropy of the hydrogel Regarding the hydrogel, the thixotropy was quantitatively evaluated using a rheometer. As the compounds, GlcN-4-CH3Pmoc and GlcN-4-CF3Pmoc were used. For the preparation of the hydrogel, a preparation method with heating was used, and a plastic syringe was used as the container (inner diameter: about 2 cm). As the aqueous solution, 100 mM HEPES-NaOH buffer (pH 7.4) containing 5.0% by volume of DMSO was used. The concentration of the compound was 1.0% by volume. It was placed on the stage of the TA instruments AR-G2 rheometer, and a 20-mm stainless steel parallel plate was brought into contact at a GAP of 1500 μm for vibration measurement. To evaluate the thixotropy, vibration measurement (1.0 rad / s) was performed at a strain of 0.1%, and the time changes of the storage modulus G' and the loss modulus G'' were measured. Then, for the purpose of solubilizing by stress, vibration measurement (1.0 rad / s) was performed at a strain of 150%. Thereafter, vibration measurement (1.0 rad / s) was performed again at a strain of 0.1%. This change in strain (from 0.1% to 150%) was repeated 3 times.

[0068] FIG. 4 is an explanatory diagram showing the measurement results of the storage modulus G' and the loss modulus G'' of GlcN-4-CH3Pmoc. FIG. 5 is an explanatory diagram showing the evaluation results of the thixotropic property of GlcN-4-CH3Pmoc. FIG. 6 is an explanatory diagram showing the measurement results of the storage modulus G' and the loss modulus G'' of GlcN-4-F3Pmoc. FIG. 7 is an explanatory diagram showing the evaluation results of the thixotropic property of GlcN-4-F3Pmoc. For both gels of GlcN-4-CH3Pmoc and GlcN-4-CF3Pmoc, at strain 0.1%, values of G'>G'' were shown, indicating characteristic behavior as a gel state. In contrast, at strain 150%, values of G'<G'' were shown, revealing that it had solated due to stress. Furthermore, as a result of returning to strain 0.1% and suppressing the stress for vibration measurement, values of G'>G'' were shown, indicating the behavior of gelling again. These results more quantitatively demonstrate that the gels of GlcN-4-CH3Pmoc and GlcN-4-CF3Pmoc have thixotropic properties.

[0069] 6. Demonstration Experiment of Injectable Gel A hydrogel showing thixotropic properties can be held by inhaling it into a syringe. Furthermore, from that state, when injected from the needle tip into the targeted site, it can gel at the discharged location. The following shows the experimental procedures and results demonstrating its performance.

[0070] As the compounds, GlcN-4-CH3Pmoc and GlcN-4-CF3Pmoc were used. For the preparation of the hydrogel, a preparation method with heating was used. As the aqueous solution, 100 mM HEPES-NaOH buffer (pH 7.4) containing 5.0% by volume of DMSO was used. The concentration of the compound was 1.0% by volume. 400 μL of the hydrogel was aspirated into a plastic syringe (1.0 mL, inner diameter = 0.60 cm) or a glass syringe (1.0 mL, inner diameter = 0.60 cm) equipped with an 18Gauge (inner diameter = 1.2 mm) syringe needle. The gel was quickly pushed out with a plunger into a glass screw vial (No. 02, 11.7 × 35 mm, manufactured by Maruemu). By this injection operation, shear stress was applied. The sample vial was inverted, and it was visually confirmed after a certain period of time that gelation had occurred.

[0071] Figure 8 is an explanatory diagram showing that the gel is injectable. The gels of GlcN-4-CH3Pmoc and GlcN-4-CF3Pmoc were both able to be aspirated and injected with a syringe. Also, in both cases, it was in a sol state immediately after injection, but it was observed that gelation occurred again within 10 minutes after stress release.

[0072] 7. Demonstration experiment on embedding of drugs It is also possible to mix drugs such as anticancer agents in the gel and inject them. The following shows the procedure and results of the experiment demonstrating its performance.

[0073] As the compounds, GlcN-4-CH3Pmoc and GlcN-4-CF3Pmoc were used. As the agent to be embedded in the gel, doxorubicin, an anticancer agent, was used. For the preparation of the hydrogel, a preparation method without heating was used. As the aqueous solution, 100 mM HEPES-NaOH buffer (pH 7.4) containing 5.0% by volume of DMSO was used. The concentration of the compound was 1.0% by volume, and the concentration of doxorubicin was 328 μM. 400 μL of the hydrogel was aspirated into a glass syringe (1.0 mL, inner diameter = 0.60 cm) equipped with an 18Gauge (inner diameter = 1.2 mm) syringe needle. The gel was quickly extruded with a plunger into a glass screw vial (No. 02, 11.7 × 35 mm, manufactured by Maruemu). The sample vial was inverted, and it was visually confirmed after a certain period of time that gelation had occurred.

[0074] Figure 9 is an explanatory diagram showing that the gel embedding the anticancer agent is injectable. Since doxorubicin exhibits a red color, it can be easily confirmed that it is embedded in the gel. The gels of GlcN-4-CH3Pmoc and GlcN-4-CF3Pmoc embedding doxorubicin were both aspirated and injected with a syringe. Also, in both cases, it was in a sol state immediately after injection, but it was observed that gelation occurred again within 10 minutes after stress release.

[0075] The present disclosure is not limited to the above-described embodiments, and can be realized in various configurations without departing from the gist thereof. For example, the technical features in each embodiment corresponding to the technical features in each form described in the summary of the invention can be appropriately replaced or combined in order to solve some or all of the above-described problems, or to achieve some or all of the above-described effects. Also, if the technical feature is not described as essential in this specification, it can be appropriately deleted.

Claims

1. The following general formula (1) or the following general formula (2): 【Chemical 1】 (In formula (1), X is represented by the following general formula (3).) 【Chemical 2】 (In formula (2), X and Y are each independently represented by the following general formula (3).) 【Chemical 3】 (In formula (3), R １ , R ２ , and R ３ each independently represents a hydrogen atom, a halogen atom, or an alkyl group having 1 to 4 carbon atoms.) A compound represented by the formula.

2. In the general formula (1), X is para-positioned with respect to the bonding position with the amino sugar, The compound according to Claim 1.

3. In the general formula (3), R １ , R ２ , and R ３ are a hydrogen atom or a halogen atom, The compound according to Claim 1 or Claim 2.

4. The compound according to Claim 1 or Claim 2, A solvent containing water, A hydrogel comprising the same.

5. In the hydrogel according to Claim 4, The compound is contained in an amount of 0.7% by mass or more and 10% by mass or less, A hydrogel.

6. A pharmaceutical composition comprising the hydrogel according to Claim 4.

7. Use of the compound according to Claim 1 or Claim 2 for the preparation of a hydrogel.

Images