Peroxide hydrogel and preparation method thereof

Abstract

The invention discloses peroxide hydrogel and a preparation method thereof. The peroxide hydrogel comprises peroxide and connecting molecules or salts thereof. The peroxide hydrogel is prepared by centrifugally cleaning a suspension formed by mixing a metal compound, a connecting molecule, hydrogen peroxide and an auxiliary agent in a solvent. According to the invention, an initiator and a cross-linking agent are not used, the peroxide is gelatinized by connecting molecules, the distribution is uniform, the viscoelasticity is excellent, the thixotropy is realized, the gel can be re-gelatinized after being applied in a sol state, and the slow release property is realized.

Description

Technical Field

[0001] The present invention relates to a peroxide hydrogel, and more particularly to a hydrogel consisting essentially of peroxide and a method for preparing the hydrogel without using an initiator and a crosslinking agent. Background Art

[0002] Hydrogen peroxide (H 2 O 2 ) is a reactive oxygen species and is widely used in the fields of life sciences and medicine. Its oxygen production and free radical generation functions enable it to play an important role in cell signaling, wound healing, and antibacterial disinfection. However, the strong oxidizing property of hydrogen peroxide may also cause cytotoxicity and tissue damage. Peroxides, as derivatives of hydrogen peroxide, can gradually degrade and release hydrogen peroxide under physiological conditions, thereby achieving controllable hydrogen peroxide release and reducing the risk of excessive release. This sustained-release mechanism shows application prospects in anti-microbial infection, promoting wound healing, and cancer treatment. For example, by locally releasing hydrogen peroxide in specific tissues or tumor areas, the anti-tumor therapeutic effect can be enhanced.

[0003] At present, there are many options for the implementation of peroxide in clinical applications. One way is to load the peroxide in a biocompatible gel carrier and adjust its release rate by the slow-release properties of the gel. For example, patent CN115887780B discloses an oxygen slow-release hydrogel, in which peroxide, a photoinitiator and a methacrylated gelatin solution are mixed so that the peroxide is evenly dispersed in the hydrogel substrate solution, and ultraviolet cross-linking is adopted to obtain a hydrogel material. CN113499444A discloses an oxygen slow-release material and its preparation method and medicine, wherein the peroxide is dispersed in a mixed matrix of polycaprolactone or polycaprolactone and polylactic acid. CN113908332B discloses that metal peroxide induces rapid cross-linking of a high molecular polymer grafted with phenolic hydroxyl groups to prepare a hydrogel under the action of peroxidase. However, this method has high requirements on the biocompatibility and degradability of the gel carrier, the preparation process is complicated, and the distribution of the loaded drug may be uneven. Summary of the invention

[0004] A controlled peroxide gel system that slowly releases physiologically active substances such as hydrogen peroxide and oxygen is effective for obtaining sustained therapeutic effects or reducing side effects. The present invention aims to provide a hydrogel system using peroxide as a basic building block, which does not use an initiator and a cross-linking agent, and is a viscoelastic hydrogel that is essentially composed of peroxide and a method for preparing the same.

[0005] The first aspect of the present invention provides a peroxide hydrogel comprising:

[0006] Peroxides comprising one or a combination of two or more of magnesium peroxide, calcium peroxide and zinc peroxide; and

[0007] A linker molecule or a salt thereof, wherein the linker molecule is a molecule that chelates with metal ions and has the structure: R 1 (CH(R 3 )) n R 2 ,

[0008] Among them, R 1 and R 2 The terminal groups of the connecting molecules each independently include: -N=CH(NH 2 ) 2 、-NH-C(=NH)NH 2 、-NH-C(=O)NH 2 , guanidinyl (-O-NH-C(=NH)NH 2 ), biguanide group, urea group, thiourea group, ether group, amino group, carboxyl group, -CH(COOH)NH 2 At least one functional group or derivative group in 2 The N-terminus and C-terminus are dehydrated and condensed to form a polypeptide chain;

[0009] Each R 3 are independently a hydrogen atom, a hydroxyl group, an amino group, or a carboxyl group;

[0010] The repeating unit n is 1, 2, 3, 4, 5, 6, 7 or 8.

[0011] When R 1 or R 2 -CH(COOH)NH 2 When the functional group is present, the linker molecule comprises a derivative of its N-terminus or C-terminus, for example, methylation or acetylation of the amino terminal, esterification or amidation of the carboxyl terminal, etc.

[0012] When R 1 or R 2 -CH(COOH)NH 2 When the functional group is present, the linking molecule also comprises a polypeptide chain formed by dehydration condensation of the N-terminus and the C-terminus.

[0013] In another preferred embodiment, the peroxide hydrogel is an injectable peroxide hydrogel.

[0014] In another preferred embodiment, the peroxide hydrogel is in a sol state.

[0015] In another preferred embodiment, the peroxide is formed by a substance selected from the group consisting of magnesium chloride, zinc chloride, calcium peroxide, zinc nitrate, zinc oxide, magnesium nitrate, and magnesium hydroxide.

[0016] In another preferred embodiment, the linking molecule is selected from at least one of arginine, citrulline, canavanine, glycine, lysine, histidine, proline, L-2-amino-4-guanidinobutyric acid, 1,1'-(butane-1,4-diyl)biguanide, homoarginine, arginineamide, β-guanidine propionate, guanidinoglutaric acid, 5-guanidinopentanoic acid, ARG-ARG, Nα-acetyl-L-arginine, nopaline, 2,6-diaminopimelate sodium salt, and H-ARG-ALA-OH.

[0017] In another preferred embodiment, the linker molecule is selected from: arginine, citrulline, canavanine, and 1,1'-(butane-1,4-diyl)biguanide.

[0018] There is no particular limitation on the form of the linker molecule, which may be the above-mentioned hydrate, salt, or derivative. There is no particular limitation on the absolute stereo configuration of the molecule, which may be L configuration, D configuration, or racemate.

[0019] In another preferred embodiment, the weight ratio of the peroxide to the linker molecule is 1:0.1 to 1:6, preferably 1:0.4 to 1:4.

[0020] In another preferred embodiment, the peroxide hydrogel further comprises an auxiliary agent, which can improve the stability of the gel system, adjust the pH value of the hydrogel system, and slow down the hydrolysis of peroxide in the aqueous gel.

[0021] In the present invention, the auxiliary agent is selected from: one or a combination of two or more of ammonia water, ethanolamine, diethanolamine, triethanolamine, triethylamine, and choline hydroxide, preferably ammonia water.

[0022] In another preferred embodiment, the auxiliary agent is selected from: (10%-50%, preferably 30%) ammonia water, ethanolamine.

[0023] In another preferred embodiment, the water content in the hydrogel is 10wt% to 90wt%.

[0024] The present invention integrates peroxide as a part of gel formation directly into the hydrogel network, simplifies the drug loading step, ensures the uniform distribution of peroxide in the gel, and improves the stability and uniformity of the gel system.

[0025] The peroxide hydrogel of the present invention is injectable, can be applied in a sol state, and can gel in vivo after application. Furthermore, since no light, heat, enzyme or pH change is required during the gelation process, some substances can also be encapsulated.

[0026] The second aspect of the present invention provides a method for preparing the hydrogel according to the first aspect, comprising the following steps:

[0027] 1) mixing a metal compound, a linker molecule, hydrogen peroxide, and an optional auxiliary agent in a solvent to form a suspension;

[0028] 2) The suspension is centrifuged and washed to obtain a purified hydrogel.

[0029] Wherein, the solvent is an organic solvent or a mixed solvent of water and an organic solvent, wherein the water content is 0% to 50% v / v, preferably 10% to 35%;

[0030] The metal compound is at least one of a salt, oxide or peroxide of calcium, magnesium or zinc;

[0031] The weight ratio of the linking molecule to the metal content in the metal compound is 0.1:1 to 10:1 w / w.

[0032] In another preferred embodiment, the auxiliary agent is selected from: ammonia water, ethanolamine, diethanolamine, triethanolamine, triethylamine, tetramethylammonium hydroxide, tetraethylammonium hydroxide, choline hydroxide, one or a combination of two or more thereof.

[0033] In another preferred embodiment, the metal salt is selected from at least one of calcium salt, magnesium salt and zinc salt. There is no particular limitation on the form of the metal salt, which may be the anhydrous compound, hydrate or derivative mentioned above.

[0034] Taking magnesium salt as an example, the following can be cited: anhydrous compounds of metal salts, for example, anhydrous magnesium chloride, anhydrous magnesium sulfate. Hydrates of metal salts, for example, magnesium chloride hexahydrate. Derivatives of metal salts, for example, magnesium nitrate, magnesium hydroxide, magnesium thiosulfate.

[0035] The metal oxide comprises at least one of calcium oxide, magnesium oxide and zinc oxide. The metal peroxide comprises at least one of calcium peroxide, magnesium peroxide and zinc peroxide. The metal oxide and the metal peroxide are nanoparticles with an average particle size of less than or equal to 100 nanometers. The microscopic morphology of the nanoparticles can be one-dimensional, two-dimensional or three-dimensional, for example, nanowires, nanosheets or nanospheres.

[0036] In another preferred embodiment, the metal compound is selected from the group consisting of: magnesium chloride hexahydrate, anhydrous zinc chloride, calcium peroxide, zinc nitrate hexahydrate, zinc oxide, magnesium nitrate, and magnesium hydroxide.

[0037] In another preferred embodiment, the hydrogen peroxide in step 1) refers to a hydrogen peroxide solution diluted with water, which is an aqueous solution with a mass concentration of 20% to 60%, and the weight ratio of the added volume to the metal content in the metal compound is 0.01:1 to 15:1 v / w, preferably 0.09:1 to 5:1 v / w.

[0038] In another preferred embodiment, the hydrogen peroxide in step 1) refers to a hydrogen peroxide solution diluted with water, which is an aqueous solution with a mass concentration of 25% to 35%, and the weight ratio of the added volume to the metal content in the metal compound is 0.09:1 to 5:1 v / w.

[0039] Preferably, the auxiliary agent in step 1) is selected from ammonia water and ethanolamine, and the volume ratio of the auxiliary agent to hydrogen peroxide is 0.1:1 to 5:1 v / v.

[0040] Preferably, the solvent of the suspension in step 1) is an organic solvent.

[0041] Preferably, the solvent of the suspension in step 1) is a combination of water and an organic solvent, wherein the water content is 10% to 35% v / v.

[0042] The type of solvent used in the suspension should be selected so as not to substantially affect the chemical properties or physical integrity of the raw materials.

[0043] The organic solvent is selected from the group consisting of: methanol, ethanol, isopropanol, 1,2-ethylene glycol, 1,2-propylene glycol, 1,3-propylene glycol, pyridine, acetonitrile, dimethyl sulfoxide, tetrahydrofuran, formamide, N-methylformamide, N,N-dimethylformamide, N,N-diethylformamide, and N-methylpyrrolidone, or a mixed solvent of two or more thereof.

[0044] In another preferred embodiment, the solvent is selected from one or a mixed solvent of two or more selected from methanol, water, N-methylformamide, N,N-dimethylformamide, and N-methylpyrrolidone.

[0045] In another preferred embodiment, the weight ratio of the linker molecule to the metal content in the metal compound is 0.3:1 to 5:1 w / w.

[0046] In another preferred embodiment, step 1) comprises: dissolving the metal compound in a solvent to form a first mixed solution; mixing hydrogen peroxide, a linker molecule, an auxiliary agent, and a solvent to form a second mixed solution; mixing the second mixed solution with the first mixed solution to obtain a suspension;

[0047] Alternatively, step 1) comprises: dissolving the metal compound and the linker molecule in a solvent to form a first mixed solution; mixing hydrogen peroxide, an auxiliary agent and an optional solvent to form a second mixed solution; mixing the first mixed solution and the second mixed solution to obtain a suspension;

[0048] Alternatively, step 1) comprises: dissolving the metal compound and the auxiliary agent in a solvent to form a first mixed solution; mixing hydrogen peroxide, a linker molecule and a solvent to form a second mixed solution; and mixing the first mixed solution and the second mixed solution to obtain a suspension.

[0049] In another preferred embodiment, the mixing method in step 1) is selected from one or a combination of dropwise addition, mechanical stirring, magnetic stirring, microchannel mixing, and ultrasonic mixing.

[0050] The mixing environment temperature range of step 1) is 0-90° C., and the mixing time is 1 min-24 h.

[0051] Preferably, the cleaning in step 2) is selected from a combination of centrifugation, dialysis, chromatography, ultrafiltration, etc. The selection of the cleaning method is not particularly limited, as long as the purpose of removing excess substances and purification is achieved.

[0052] This method optimizes the preparation process, reduces production costs, and improves the feasibility of large-scale industrial production. The peroxide integrated into the hydrogel network has significantly enhanced stability and can avoid activity loss caused by environmental factors.

[0053] The third aspect of the present invention provides a functional peroxide hydrogel, comprising the peroxide hydrogel described in the first aspect; and

[0054] Functional factors that promote peroxide oxygen production or the generation of active free radicals in hydrogels.

[0055] The functional factors can be listed as follows:

[0056] Functional factors that promote oxygen production, such as catalase, manganese ions, manganese oxide, manganese dioxide, titanium dioxide, zinc sulfide, platinum, silver, palladium, and ruthenium.

[0057] Functional factors that promote the generation of active free radicals, such as transition metals (iron, copper, cobalt, aluminum), transition metal oxides (iron oxyhydroxide, copper oxide, cobalt oxide), transition metal salts (ferric chloride, copper sulfate, cobalt chloride), and transition metal complexes (ferrocene, carbonyl iron, ferric citrate).

[0058] In another preferred embodiment, the functional factor is selected from: at least one or a combination of two or more of catalase, manganese dioxide, iron oxide, iron oxyhydroxide, copper oxide, cobalt oxide, iron chloride, copper sulfate, cobalt chloride, ferrocene, carbonyl iron, and ferric citrate.

[0059] In another preferred embodiment, the functional factor is selected from: manganese dioxide, catalase, copper chloride, and ferric oxyhydroxide.

[0060] In another preferred embodiment, the weight ratio of the functional factor to the peroxide is 0.001-0.5 g:1 g.

[0061] The fourth aspect of the present invention provides a method for preparing the functional peroxide hydrogel according to the third aspect, comprising the step of mixing the functional factor with the purified peroxide hydrogel.

[0062] The mixing method is selected from: dropwise addition, mechanical stirring, magnetic stirring, microchannel mixing, ultrasonic mixing, or a combination of several methods.

[0063] The mixing environment temperature ranges from 0 to 90° C., and the mixing time ranges from 1 minute to 24 hours.

[0064] After mixing, the mixture is allowed to stand to obtain a functional peroxide hydrogel. The range of the standing environment temperature is 0 to 60° C., and the standing time is 1 to 24 hours.

[0065] A fifth aspect of the present invention provides a peroxide hydrogel powder obtained by drying the peroxide hydrogel described in the first aspect and removing the solvent.

[0066] The peroxide hydrogel powder comprises peroxide and linker molecules. In another preferred embodiment, the powder further comprises an auxiliary agent.

[0067] In another preferred embodiment, the peroxide hydrogel powder further contains a stabilizer. The stabilizer can protect the hydrogel after removing water, maintain the original microscopic skeleton structure, and protect the gelling functional group, so that it can be re-gelled after being redissolved in water. The stabilizer is selected from one or a combination of citric acid, trisodium citrate, trimagnesium citrate, and tricalcium citrate.

[0068] The peroxide hydrogel powder of the present invention can be re-gelled after being re-dissolved in water, and thus can be conveniently applied to the wound site.

[0069] The sixth aspect of the present invention provides a method for preparing the peroxide hydrogel powder described in the fifth aspect:

[0070] Mixing the peroxide hydrogel of the first aspect, an optional stabilizer and a solvent;

[0071] After gelation, the peroxide hydrogel powder is obtained by drying to remove the solvent.

[0072] In another preferred embodiment, the stabilizer is selected from one or a combination of citric acid, trisodium citrate, trimagnesium citrate, and tricalcium citrate.

[0073] Preferably, the amount of stabilizer added to the powder is 0 to 3 g / g peroxide.

[0074] Preferably, the solvent is removed by spray drying, supercritical fluid drying or freeze drying.

[0075] A seventh aspect of the present invention provides a functional peroxide hydrogel powder obtained by the following two methods:

[0076] Drying the functional peroxide hydrogel to remove the solvent to obtain functional peroxide hydrogel powder; or

[0077] The peroxide hydrogel powder is mixed with the functional factor powder to obtain the functional peroxide hydrogel powder.

[0078] The functional peroxide hydrogel, solvent, peroxide hydrogel powder and functional factors are as described above.

[0079] Preferably, the powder further contains a stabilizer. The stabilizer can protect the hydrogel after removing water, maintain the original microscopic skeleton structure, and protect the gelling functional group, so that it can be re-gelled after being redissolved in water. The stabilizer is selected from one or a combination of citric acid, trisodium citrate, trimagnesium citrate, and tricalcium citrate.

[0080] Preferably, the amount of stabilizer added to the powder is 0 to 3 g / g peroxide.

[0081] Preferably, the solvent is removed by spray drying, supercritical fluid drying or freeze drying.

[0082] Preferably, the mixing is selected from one or a combination of mechanical stirring mixing and air flow mixing.

[0083] In an eighth aspect of the present invention, there is provided a use of the peroxide hydrogel described in the first aspect, the functional peroxide hydrogel described in the third aspect, the peroxide hydrogel powder described in the fifth aspect or the functional peroxide hydrogel powder described in the seventh aspect, for releasing hydrogen peroxide or oxygen; for use as a drug or protease carrier; or for preparing drugs for resisting microbial infection, promoting wound healing and treating cancer.

[0084] The present invention does not use any crosslinking agent such as photocrosslinking or chemical crosslinking, but gels the peroxide by connecting molecules, thereby preparing a peroxide hydrogel with a small amount of additives and excellent viscoelasticity. In addition, the present invention does not use organic compounds that may have biosafety hazards. Furthermore, the gel system of the present invention has thixotropy and can gel again in the body after being applied in a sol state, so it can be applied by a syringe without damaging the gel properties such as slow release. In addition, application by spraying, dripping, or coating is also possible.

[0085] Compared with existing loading technology, peroxide is directly involved in gel formation to ensure its uniform distribution in gel matrix, avoids local concentration to be too high and causes uneven release. In addition, as a part for gel, the stability of peroxide is significantly improved, avoids the activity loss caused by environmental factors. Moreover, due to the gelation without the need for heating or ultraviolet light, for example, the inner sealing of protease becomes possible for the material with weak heat or light reaction, and the possibility of denaturation is reduced when the inner sealing is unstable. In addition, other functional components can also be further introduced in peroxide hydrogel to reach the purpose of promoting the release of oxygen or the generation of active free radicals.

[0086] The peroxide hydrogel of the present invention provides a feasible technical solution for the fields of antibacterial disinfection, wound repair and cancer treatment, and has good application prospects.

[0087] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equal or similar purpose. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0088] Figure 1 Photograph showing the format of the peroxide hydrogel of the present invention.

[0089] Figure 2 The rheological test results of the peroxide hydrogel of the present invention are shown.

[0090] Figure 3 The rheological test results of the peroxide hydrogel of the present invention are shown.

[0091] Figure 4 Photograph showing the transformation of the gel powder of the present invention from a sol state to a gel state after being redissolved in water.

[0092] Figure 5 It shows that after the peroxide hydrogel of the present invention is squeezed out by a syringe, the drawn pattern still maintains its original shape.

[0093] Figure 6 The hydrogel system of the present invention is shown to have a sustained release effect on hydrogen peroxide.

[0094] Figure 7 The effect of oxygen generation by the hydrogel system of the present invention is shown.

[0095] Figure 8 The results show that the hydrogel system of the present invention promotes the healing of mouse wounds. DETAILED DESCRIPTION

[0096] the term

[0097] Unless otherwise defined herein, all terms (including technical and scientific terms) used have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0098] The term "peroxide" refers to a compound containing a peroxy group (-OO-) and containing one or more peroxy bonds in its molecular structure.

[0099] The term "hydrogel" refers to a three-dimensional network structure formed by cross-linked molecules that can absorb and retain a large amount of water in water and maintain a soft, elastic and shaped gel state. Hydrogels are usually formed by chemical or physical cross-linking of molecules, and their network structure allows water molecules to enter and exit freely while maintaining the interconnection between the cross-linked molecular chains.

[0100] The term "metal content of a metal salt" refers to the weight of the metal contained in the metal salt compound.

[0101] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods for the unrecorded specific conditions in the following examples are usually performed under normal conditions or according to the conditions recommended by the manufacturer. Unless otherwise indicated, percentages and parts are weight percentages and weight parts.

[0102] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0103] Materials: magnesium chloride hexahydrate, magnesium hydroxide, anhydrous zinc chloride, zinc nitrate hexahydrate, 80% methanol (methanol-water mixture, volume fraction), 30% hydrogen peroxide (mass fraction), 30% ammonia water (mass fraction), arginine, citrulline, canavanine, 1,1'-(butane-1,4-diyl) biguanide sulfate, catalase, trisodium citrate, triethanolamine, purchased from Sigma-Aldrich;

[0104] Calcium peroxide nanoparticles (50 mmol / L), zinc oxide nanodots (100 mmol / L), and manganese dioxide nanosheets (20 mg / mL) were purchased from Qian Yang Bio-Tech (Shanghai) Limited.

[0105] Example 1

[0106] Dissolve 1.02g of magnesium chloride hexahydrate in a flask containing 10mL of 80% methanol. In addition, add 40mg of arginine, 0.5mL of 30% hydrogen peroxide and 1.5mL of 30% ammonia water to a beaker containing 40mL of 80% methanol and mix well. Under magnetic stirring, quickly add the solution in the beaker to the flask, and continue the reaction at a constant temperature for 3 hours. Collect the suspension and separate the product by high-speed centrifuge. The product is redissolved in 20mL of water, and the supernatant is removed again by centrifugal force of 10000g to obtain peroxide hydrogel.

[0107] Figure 1 Photographs showing the patterns of hydrogels. Figure 2 The rheological test results of the hydrogel are shown. The results show that the prepared product has typical hydrogel properties. It maintains the gel state under low shear and shows obvious viscoelastic changes during the gel-sol transition process.

[0108] Example 2

[0109] Dissolve 1.36g of anhydrous zinc chloride and 487mg of arginine in a flask containing 20mL of 80% methanol. In addition, add 1mL of 30% hydrogen peroxide and 3.5mL of 30% ammonia water to a beaker containing 45mL of 80% methanol and mix well. Under mechanical stirring, quickly add the solution in the flask to the beaker, and continue the reaction at a constant temperature for 20 minutes. Collect the suspension and use a high-speed centrifuge to separate the product. The product is redissolved in 20mL of water and mixed. After standing at room temperature overnight, the supernatant is separated again with a centrifugal force of 20000g to obtain a peroxide hydrogel.

[0110] Figure 3 The rheological test results of peroxide hydrogel are shown. The results show that the prepared product has typical hydrogel properties. It maintains the gel state under low shear and shows obvious viscoelastic changes during the gel-sol transition process.

[0111] Example 3

[0112] Regulation of hydrogel strength

[0113] The preparation method of the hydrogel is the same as that of Example 2, except that the amount of arginine added is adjusted to 150 mg (Group 1), 250 mg (Group 2), 450 mg (Group 3) and 600 mg (Group 4). The storage modulus (G') of the obtained hydrogel under 0.1% shear strain is shown in the table below. The results show that the content of the linking molecule in the hydrogel significantly affects the strength of the hydrogel, and increasing the content of the linking molecule can improve the strength of the hydrogel.

[0114] Group 1 2 3 4 G′(Pa) 961 4347 9264 15648

[0115] Example 4

[0116] Dissolve 10 mL of calcium peroxide nanoparticles and 18 mg of citrulline in a flask containing 10 mL of ultrapure water. In addition, mix 0.1 mL of 30% hydrogen peroxide and 0.5 mL of 30% ammonia solution. Under mechanical stirring, slowly add the mixed solution dropwise to the flask and continue the reaction at a constant temperature for 1 hour. Collect the solution and separate the product by centrifugation using a high-speed centrifuge. The product is redissolved in 10 mL of water and mixed. After standing at room temperature overnight, remove the supernatant again with a centrifugal force of 20,000 g, and add an appropriate amount of water to the centrifuged substrate to swell to obtain a peroxide hydrogel.

[0117] Example 5

[0118] Dissolve 2.975g zinc nitrate hexahydrate and 487mg canavanine in a flask containing 40mL of 80% methanol. Add 1mL of 30% hydrogen peroxide, 1.7mL of 30% ammonia water and 0.3mL of ethanolamine to a beaker containing 20mL of 80% methanol and mix well. Under mechanical stirring, use a syringe pump to add the solution in the beaker dropwise to the flask at a rate of 1ml / min, and continue the reaction at room temperature for 2 hours. Collect the reaction solution and separate the product by centrifugation using a high-speed centrifuge. The product is redissolved in 10mL of water and then dialyzed in ultrapure water for 24 hours in an ice water bath to obtain a peroxide hydrogel.

[0119] Example 6

[0120] Dissolve 10 mL of zinc oxide nanodots and 98 mg of 1,1'-(butane-1,4-diyl) biguanide sulfate in a flask containing 10 mL of ultrapure water. In addition, mix 2 mL of 30% hydrogen peroxide and 1 mL of 30% ammonia solution. Under mechanical stirring, the mixture was slowly added dropwise to the flask and continued to react for 0.5 hours in an ice-water bath. Collect the reaction solution and separate the product by centrifugation using a high-speed centrifuge. The centrifuged substrate was redissolved in 10 mL of water and then dialyzed in ultrapure water for 24 hours in an ice-water bath to obtain a peroxide hydrogel.

[0121] 20 mL of the manganese dioxide nanosheet solution was added to the above peroxide hydrogel, and mixed evenly with a stirring rod, and then allowed to stand at room temperature for 1 hour to obtain a functional peroxide hydrogel.

[0122] Example 7

[0123] Dissolve 1.734g of magnesium chloride hexahydrate in a flask containing 30mL of N-methylformamide. Add 130mg of arginine to 6mL of ultrapure water and dissolve it completely, then mix it evenly with 0.92mL of 30% hydrogen peroxide and 3mL of 30% ammonia water. Under magnetic stirring, the mixture was quickly added to the flask and the reaction was continued for 5 hours under ice-water bath conditions. The suspension was collected and the product was separated by centrifugation using a high-speed centrifuge. The product was redissolved in 30mL of water, and the supernatant was removed again with a centrifugal force of 10000g. The centrifuged substrate was dispersed in 10mL of water to obtain a peroxide hydrogel.

[0124] 200 mg of citric acid was completely dissolved in 1 mL of ultrapure water, and then the above peroxide hydrogel was added and mixed evenly with a stirring rod. Then triethanolamine was added to adjust the pH value of the system to 8.8, and 5 mg of catalase was added. After mixing evenly, the mixture was allowed to stand for 2-4 hours to obtain a functional peroxide hydrogel.

[0125] Example 8

[0126] Dissolve 0.48g of magnesium nitrate in a flask containing 12mL of N,N-dimethylformamide. Add 50mg of arginine to 2.5mL of ultrapure water and dissolve it completely, then mix it evenly with 0.35mL of 30% hydrogen peroxide and 1.14mL of 30% ammonia water. Under magnetic stirring, quickly add the mixture to the flask and continue the reaction for 2 hours under ice-water bath conditions. Collect the suspension and separate the product by high-speed centrifugation. The product is redissolved in 30mL of water, and the supernatant is removed again by centrifugal force of 10000g. The centrifugal substrate is dispersed in 10mL of water to obtain a peroxide hydrogel.

[0127] Dissolve 280 mg of trisodium citrate completely in 1 mL of ultrapure water, then add the above peroxide hydrogel and mix evenly with a stirring rod. After standing for half an hour to allow it to gel, freeze the hydrogel at -20°C for 12 hours, using slow freezing. Then place it in a freeze dryer for freeze drying, with a vacuum degree of 25 Pa for sublimation drying and 1 Pa for analytical drying. The heating plate temperature during analytical drying is 20°C, and the hydrogel powder is obtained.

[0128] Figure 4 Photograph showing the transformation of hydrogel powder from sol to gel after being redissolved in water.

[0129] Example 9

[0130] The preparation method of the hydrogel is the same as that of Example 7. After the preparation is completed, the hydrogel is placed in a freezer at -20°C for 12 hours, using slow freezing. Then, it is freeze-dried in a freeze dryer, with a vacuum degree of 25Pa for sublimation drying and a vacuum degree of 1Pa for analytical drying. The heating plate temperature during analytical drying is 20°C, and the functional gel powder is obtained.

[0131] Example 10

[0132] The preparation method of the hydrogel powder is the same as that of Example 8. After the preparation is completed, 1 g of the hydrogel powder is completely mixed with 10 mg of copper chloride powder to obtain a functional hydrogel powder.

[0133] Embodiment 11

[0134] Dissolve 0.5g of magnesium hydroxide in a flask containing 3mL of 30% ammonia water and 30mL of N-methylpyrrolidone. Add 130mg of arginine to 6mL of ultrapure water and dissolve it completely, then mix it evenly with 0.92mL of 30% hydrogen peroxide. With the assistance of ultrasound, the mixed solution is quickly added to the flask. Then use magnetic stirring and continue the reaction for 5 hours under ice-water bath conditions. Collect the reaction solution and separate the product by centrifugation in a high-speed centrifuge. The product is redissolved in 30mL of water, and the supernatant is removed again with a centrifugal force of 10000g. The centrifuged substrate is dispersed in 10mL of water to obtain a peroxide hydrogel.

[0135] 142 mg of trisodium citrate was completely dissolved in 1 mL of ultrapure water, and then added to the above peroxide hydrogel and mixed evenly with a stirring rod. Ammonia water was then added to adjust the pH value of the system to 8.8, and then 8 mg of ferric oxyhydroxide was added, mixed evenly, and then allowed to stand for 2-4 hours to obtain a functional peroxide hydrogel.

[0136] Test Example 1

[0137] Sol-Gel Transition Characteristics

[0138] According to the finished product prepared by the present invention (according to Example 2), the hydrogel in the sol state is added with red dye, then placed in a syringe, and the fluid is pressed out from the needle to draw a pattern, and then left to stand for its gelation transformation. Figure 5 The hydrogel shows the gel state after being pressed out, still maintaining the original pattern shape.

[0139] Test Example 2

[0140] Sustained release of hydrogen peroxide from peroxide hydrogel

[0141] According to the finished product prepared according to the present invention (according to Example 1), 0.1 mL of the hydrogel was added to 1 mL of ultrapure water and placed in a dialysis bag. The dialysis bag was placed in 20 mL of Tris-HCl buffer (pH 7.4), and the concentration of hydrogen peroxide in the buffer was sampled and tested at different time points. Figure 6 Demonstrate the sustained release effect of hydrogel on hydrogen peroxide.

[0142] Test Example 3

[0143] Oxygen generation effect of peroxide hydrogel

[0144] According to the finished product prepared according to the present invention (according to Example 7), 5 mL of the hydrogel was added to a beaker containing 20 mL of Tris-HCl buffer (pH 7.4). Under magnetic stirring, the dissolved oxygen in the buffer was recorded at different time points using a portable dissolved oxygen analyzer. Figure 7 Demonstrating the effect of oxygen generation by hydrogel.

[0145] Test Example 4

[0146] Subcutaneous administration of peroxide hydrogel to mice

[0147] According to the finished product prepared by the present invention (according to Example 7), the hydrogel in the sol state is placed in a syringe. Kunming mice are anesthetized with isoflurane, and after the back hair is removed, the sample is injected subcutaneously. The hydrogel of the present invention is converted into a gel state in vivo after being administered by a syringe.

[0148] Test Example 5

[0149] Application of hydrogel powder to the wound surface of mice

[0150] Kunming mice were anesthetized with isoflurane, and after depilation of the back, a portion of the skin with a diameter of about 10 mm was removed using surgical scissors to produce a wound model with full-thickness skin loss. According to the finished product prepared by the present invention (according to Example 9), the wound was moistened with physiological saline, and then the hydrogel powder was evenly covered on the wound to absorb water and turn into a gel state. New hydrogel powder was replaced at the wound site every day. During the 14-day observation period, the changes in the size of the wound were recorded. In addition, a control group without treatment was set up for comparison. The changes in the area of ​​the wounds in each group are shown in the figure. Figure 8 As shown, the hydrogel system was shown to promote healing in a mouse wound model.

[0151] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A peroxide hydrogel, characterized in that The hydrogel comprises: Peroxide, including one or a combination of two or more of magnesium peroxide, calcium peroxide and zinc peroxide; A linker molecule or a salt thereof, wherein the linker molecule is a molecule that chelates with metal ions and has the structure: R1(CH(R3)) n R2, Wherein, R1 and R2 are terminal groups connecting molecules, and each independently comprises: at least one functional group or derivative group selected from -N=CH(NH2)2, -NH-C(=NH)NH2, -NH-C(=O)NH2, guanidoxy (-O-NH-C(=NH)NH2), biguanidyl, urea, thiourea, ether, amino, carboxyl, -CH(COOH)NH2, or the N-terminus and C-terminus of -CH(COOH)NH2 are dehydrated and condensed to form a polypeptide chain; Each R3 is independently a hydrogen atom, a hydroxyl group, an amino group, or a carboxyl group; The repeating unit n is 1, 2, 3, 4, 5, 6, 7 or 8.

2. The hydrogel according to claim 1, characterized in that The linking molecule is selected from at least one of arginine, citrulline, canavanine, glycine, lysine, histidine, proline, L-2-amino-4-guanidinobutyric acid, 1,1'-(butane-1,4-diyl)biguanide, homoarginine, arginineamide, β-guanidine propionate, guanidinoglutaric acid, 5-guanidinopentanoic acid, ARG-ARG, Nα-acetyl-L-arginine, nopaline, 2,6-diaminopimelate sodium salt, and H-ARG-ALA-OH.

3. The hydrogel according to claim 1, characterized in that The weight ratio of the peroxide to the linking molecule is 1:0.1 to 1:

6.

4. The hydrogel according to claim 1, wherein The peroxide hydrogel further comprises an auxiliary agent, which is selected from: one or a combination of two or more of ammonia water, ethanolamine, diethanolamine, triethanolamine, triethylamine, and choline hydroxide, preferably ammonia water.

5. The method for preparing the hydrogel according to claim 1, characterized in that: The preparation method comprises the following steps: 1) mixing a metal compound, a linker molecule, hydrogen peroxide, and an optional auxiliary agent in a solvent to form a suspension; 2) The suspension is centrifuged and washed to obtain a purified hydrogel. Wherein, the solvent is an organic solvent or a mixed solvent of water and an organic solvent, wherein the water content is 0% to 50% v / v; The metal compound is at least one of a salt, oxide or peroxide of calcium, magnesium or zinc; The weight ratio of the linking molecule to the metal content in the metal compound is 0.1:1 to 10:1 w / w.

6. The preparation method according to claim 5, characterized in that: The weight ratio of the linking molecule to the metal content in the metal compound is 0.3:1 to 5:1 w / w.

7. A functional peroxide hydrogel, characterized in that: The functional peroxide hydrogel comprises the peroxide hydrogel according to any one of claims 1 to 4; and Functional factors that promote peroxide oxygen production or the generation of active free radicals in hydrogels.

8. The functional peroxide hydrogel according to claim 7, characterized in that The functional factor is selected from: at least one or a combination of two or more of catalase, manganese dioxide, iron oxide, iron oxyhydroxide, copper oxide, cobalt oxide, iron chloride, copper sulfate, cobalt chloride, ferrocene, carbonyl iron, and ferric citrate.

9. A peroxide hydrogel powder obtained by drying the peroxide hydrogel according to any one of claims 1 to 4 and removing the solvent.

10. A functional peroxide hydrogel powder obtained by the following two methods: Drying the functional peroxide hydrogel according to claim 7 to remove the solvent to obtain functional peroxide hydrogel powder; or The peroxide hydrogel powder according to claim 9 is mixed with functional factor powder to obtain functional peroxide hydrogel powder.

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