A kind of natural polymer hydrogel of enzyme-catalyzed disulfide bond cross-linking and its preparation method

An enzyme-catalyzed disulfide bond, natural polymer technology, applied in the field of medical biomaterials, can solve the problems of reduced enzyme activity, poor hydrogel uniformity, affecting the activity of loaded drugs, etc., achieving fast cross-linking speed, simple operation, excellent Effects of Biocompatibility and Biodegradability

Active Publication Date: 2019-03-01
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But by directly adding hydrogen peroxide to activate horseradish peroxidase, often because hydrogen peroxide is excessive, cause horseradish peroxidase to form inert intermediate, cause enzyme activity to reduce (K.J.aynton, J.K.ewtra, N.iswas and K.E.Taylor., Biochim. Biophys.Acta.1994,1206,272), or the formed hydrogel has poor uniformity, which may even affect the activity of loading drugs, thus limiting the application of enzyme-catalyzed cross-linked hydrogels as drug carriers to a certain extent

Method used

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  • A kind of natural polymer hydrogel of enzyme-catalyzed disulfide bond cross-linking and its preparation method
  • A kind of natural polymer hydrogel of enzyme-catalyzed disulfide bond cross-linking and its preparation method
  • A kind of natural polymer hydrogel of enzyme-catalyzed disulfide bond cross-linking and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Preparation of thiol-grafted hyaluronic acid macromolecule (HA-SH).

[0040] Sodium hyaluronate (2 g, 5 mmol) was weighed and dissolved in 100 mL of double distilled water, and stirred at room temperature with a magnetic stirrer until completely dissolved. Weigh N-hydroxysuccinimide (NHS) (2.3g, 20mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC.HCL) (3.832g , 20 mmol) was added to the above-mentioned completely dissolved sodium hyaluronate solution, the pH value of the solution was adjusted to about 5.4, and activated at room temperature for 1 h. Cystamine dihydrochloride (3.372g, 15mmol) was weighed and added to the above activated solution, and stirred at room temperature for 48h. The reaction solution was transferred into a dialysis bag (MWCO3500) and dialyzed with twice distilled water for 24 hours. Weigh D,L-dithiothreitol (DTT) (4.6275g, 30mmol) and add it to the above reaction solution that has been dialyzed for 24h, adjust the pH value ...

Embodiment 2

[0042] Preparation of chitosan macromolecule grafted with mercapto groups (CS-SH)

[0043] Weigh chitosan (0.5 g) and dissolve it in 50 mL of double distilled water, add 2 mL of 1 mol / L hydrochloric acid, stir at room temperature with a magnetic stirrer until completely dissolved, and adjust the pH to 6 with 1 mol / L NaOH. Weigh N-acetylcysteine-L-amino acid (NAC) (2.3563g, 14.4mmol), N-hydroxysuccinimide (NHS) (2.0g, 17.3mmol), 1-(3-dimethylaminopropyl Base)-3-ethylcarbodiimide hydrochloride (EDC.HCL) (3.5 g, 18.3 mmol) was dissolved in 6 mL of N,N-dimethylformamide (DMF). The above DMF mixed solution was added dropwise to the completely dissolved chitosan solution, the pH value of the solution was kept at about 6.0, and the reaction was stirred at room temperature for 24 hours. The reaction solution was transferred into a dialysis bag (MWCO3500) and dialyzed with twice distilled water for 24 hours. Weigh D,L-dithiothreitol (DTT) (2.31g, 15mmol) and add it to the above react...

Embodiment 3

[0045] Preparation of polyglutamic acid macromolecules grafted with thiol groups (PGA-SH)

[0046] Sodium polyglutamate (0.3 g) was weighed and dissolved in 30 mL of double distilled water, and stirred at room temperature with a magnetic stirrer until completely dissolved. Weigh N-hydroxysuccinimide (NHS) (0.2624g, 2.28mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC.HCL) (1.14 g, 5.94 mmol) was added to the above-mentioned completely dissolved sodium polyglutamate solution, the pH value of the solution was adjusted to about 6.0, and activated at room temperature for 1 h. Cysteamine hydrochloride (0.675g, 5.94mmol) was weighed and added to the above activated solution, and stirred at room temperature for 24h. The reaction solution was transferred into a dialysis bag and dialyzed with 1 mM hydrochloric acid for 1 day, 1 wt% NaCl solution for 1 day, and 1 mM hydrochloric acid for 1 day. After the dialysis, the reaction solution was quickly frozen with li...

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PUM

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Abstract

The invention provides enzyme-catalyzed disulfide bond-crosslinked natural polymer hydrogel and a preparation method thereof. The hydrogel comprises a three-dimensional net structure composed of crosslink bonds of cystamine or N,N-diacetyl-L-cystine; under the conditions that horseradish peroxidase is used as a catalyst and a phenol compound is used as an enzyme substrate, sulfydryl-grafted natural polymers are rapidly oxidized to form disulfide bonds, so that the hydrogel is obtained. According to the invention, raw materials are safe in source and environment-friendly, the product has excellent biocompatibility; reaction conditions are mild, and the operation is simple; the system is not required to be additionally added with hydrogen peroxide, and physiological activity of a loaded drug or cells is completely guaranteed.

Description

technical field [0001] The invention belongs to the field of medical biomaterials. Background technique [0002] Hydrogel is a functional polymer material with a three-dimensional network structure obtained through moderate cross-linking. It has high water content and flexible natural characteristics, which makes it have good biocompatibility and is similar to natural Extracellular matrix, which can effectively reduce tissue irritation and cell adhesion (Park, H.; Park, K., Pharm. Res. 1996, 13, 1770). More importantly, the porous structure and high water content of hydrogel make it very suitable for embedding biologically active substances, such as cells, pharmacodynamic proteins and peptide drugs, unlike other release systems (microparticles, emulsions, etc.) The activity of cells and proteins may be damaged during the preparation process, and the preparation of the hydrogel is carried out under milder conditions, which is beneficial to preserve the activities of cells an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/26C12P13/02
Inventor 彭志平佘英奇李义赵玉莹谢标明
Owner NANCHANG UNIV
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