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Novel biological material with controlled release of nitrogen monoxide under catalysis of enzyme, as well as preparation method and application thereof

A technology of nitric oxide and biological materials, which can be applied to medical preparations without active ingredients, medical preparations containing active ingredients, and drug combinations, etc., can solve the problems that affect the effective utilization of NO in materials and the easy aggregation of NO donors. To achieve the effect of promoting wound healing, good biocompatibility, and promoting skin repair

Active Publication Date: 2013-10-09
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the difference in polarity, NO donors tend to aggregate, resulting in burst release
In addition, the hydrophobic matrix material restricts the migration of water molecules and affects the effective utilization of NO inside the material.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1: Synthesis of chitosan macromolecule CS-NO with enzyme-catalyzed controlled release of nitric oxide

[0013] 0.5 grams of chitosan with a viscosity-average molecular weight of 50,000 and a degree of deacetylation of 90% were stirred and dispersed in 50 milliliters of water, and pentynoic acid was added to stir evenly, wherein the mol ratio of chitosan repeating units to pentynoic acid was: 1: 0.2 . If the chitosan still cannot be completely dissolved, add 1 mol / liter of hydrochloric acid solution dropwise until the chitosan is just completely dissolved, and add 1-(3-dimethylaminopropyl)-3-ethylcarbodiol to the system in an ice bath. Imine hydrochloride EDC, the molar ratio of pentynoic acid and EDC is 1:3, react for 24 hours, the product is dialyzed in the mixed solution of 5 mmol / L hydrochloric acid and 1% sodium chloride for 72 hours, and then in 1 mL Dialyze in mol / L hydrochloric acid for 48 hours, and finally dialyze in aqueous solution at 4°C for 72 hour...

Embodiment 2

[0015] Example 2: Preparation of CS-NO material with enzyme-catalyzed controlled release of nitric oxide

[0016] Preparation of CS-NO solution

[0017] Disperse 0.15 g of CS-NO in 10 ml of water and stir until completely dissolved to obtain a CS-NO solution with a concentration of 15 mg / ml.

[0018] Preparation of CS-NO thin film

[0019] CS-NO was prepared as a 15% aqueous solution, and 150 microliters of the solution was coated on a cover glass with a diameter of 1 cm, and dried naturally. Soak the dried CS-NO membrane in 0.01 mol / L PBS buffer solution to obtain the CS-NO membrane.

[0020] Preparation of CS-NO porous scaffold

[0021] Dissolve chitosan with a viscosity-average molecular weight of 600,000 and a degree of deacetylation of 90% in 3% acetic acid and stir until completely dissolved, neutralize it with 1M sodium hydroxide to a pH of about 6, and prepare 10 mg of CS-NO / ml of aqueous solution, mixed according to CS:CS-NO mass ratio=5:1, stirred evenly, freeze...

Embodiment 3

[0024] Example 3: Local injection of CS-NO solution for the treatment of lower limb ischemia in diabetic mice

[0025] C57 mice were used, and the experimental animals were divided into three groups, namely saline group, CS group and CS-NO group, with 6 animals in each group. The lower limb ischemia model was established by ligation and isolation of the femoral artery of the mouse lower limbs. A certain concentration of CS-NO solution was injected into the muscles around the ligation, with a total amount of 50 microliters per mouse. Angiogenesis was observed by angiography after operation. The severity of ischemia and degree of necrosis in the CS-NO group were significantly lighter than those in the other two groups. And in 7, 14, 21, 28 days were collected. Immunofluorescence staining was used to analyze the distribution of small blood vessels and capillaries in muscle tissue. The release of NO effectively promoted angiogenesis, and the density of blood vessels was signifi...

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PUM

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Abstract

The invention discloses a novel biological material with controlled release of nitrogen monoxide under catalysis of an enzyme, as well as a preparation method and application thereof. The novel biological material with the controlled release of the nitrogen monoxide under the catalysis of the enzyme is prepared by connecting a nitrogen monoxide donor compound with stable glycosylation with a natural polymer, namely chitosan through covalent bonds. The material has good processability and can be prepared into injectable solutions, thin films, porous supports, electrostatic spinning fiber films and other various products. The CS-NO and a composite material thereof can be used for treating diabetic lower limb ischemia, skin injures and myocardial infarction disease. As the nitrogen monoxide can be released in a controlled manner according to requirements, the material shows a great effect in treatment.

Description

technical field [0001] This patent relates to a method for preparing biological materials with enzyme-catalyzed and controlled release of nitric oxide, as well as medical products processed from such materials and their applications in ischemic disease treatment, skin wound healing and cardiovascular implantation devices Applications. Background technique [0002] Cardiovascular disease has now become one of the major diseases that threaten human health, ranking first in both morbidity and mortality among various diseases, and is recognized by the medical community as the "number one killer" that endangers human health. According to the "China Cardiovascular Disease Report 2011" data released by the Cardiovascular Disease Prevention and Research Center of the Ministry of Health, the number of cardiovascular disease patients in my country is about 230 million, which is equivalent to 2 out of every 10 adults. High medical expenses become a huge economic burden. [0003] Nitr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/08C08J5/18C08J9/28D04H1/42A61K47/48A61K31/706A61P17/02A61P9/10A61L31/04A61L31/16
Inventor 赵强孔德领张计敏王淑芳
Owner NANKAI UNIV
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