Composite agarose microsphere and preparation method and application thereof

A technology of agarose microspheres and agarose, which is applied in the field of materials, can solve the problems of short drug action time, large toxic and side effects in the human body, poor biological stability, etc., and achieve the effect of easy promotion and application, cheap and large coating volume

Active Publication Date: 2019-07-12
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is: the nitric oxide donor drug has shortcomings such as short drug action time, poor biological stability, large toxic and side effects on the human body, lack of targeting and tissue specificity, etc.

Method used

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  • Composite agarose microsphere and preparation method and application thereof
  • Composite agarose microsphere and preparation method and application thereof
  • Composite agarose microsphere and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A composite agarose microsphere of the present invention, comprising hydroxyurea, β-D-glucose oxidoreductase and horseradish peroxidase, coated with hydroxyurea, β-D-glucose oxidoreductase and horseradish peroxidase Form complex agarose microspheres in agarose. figure 1 It is a schematic diagram of agarose microspheres coated with cascade enzymes, and its preparation method comprises the following steps:

[0036] (1) Prepare the organic phase: Mix 18 mL of toluene, 7 mL of chloroform and 0.3 mL of sorbitan fatty acid ester to form the organic phase. Add the above organic phase into a 100 mL flask and preheat to 55 °C at 1000 rpm.

[0037] (2) Preparation of water phase: Take 5 ml of water and add 84 mg of agarose, heat to dissolve, and obtain an agarose solution. When the temperature of the agarose solution dropped to 55°C, add 0.1 mL of β-D-glucose oxidoreductase solution with a pH of 7.0 and a concentration of 1 mg / mL and 0.5 mL of a peppery solution with a concentr...

Embodiment 2

[0040] A composite agarose microsphere of the present invention, comprising hydroxyurea, β-D-glucose oxidoreductase and ferric iron tetroxide nanoparticles, coated with hydroxyurea, β-D-glucose oxidoreductase and ferric iron tetroxide nanoparticles Form complex agarose microspheres in agarose. Its preparation method comprises the following steps:

[0041] (1) Prepare the organic phase: Mix 18 mL of toluene, 7 mL of chloroform and 0.3 mL of sorbitan fatty acid ester to form the organic phase. Add the above organic phase into a 100 mL flask and preheat to 55 °C at 1000 rpm.

[0042](2) Preparation of the water phase: Take 5 ml of water and add 84 mg of agarose, heat to dissolve to obtain an agarose solution. When the temperature of the agarose solution dropped to 55°C, add 0.1 mL of β-D-glucose oxidoreductase solution with a pH of 7.0 and a concentration of 1 mg / mL and 0.5 mL of a solution of β-D-glucose oxidoreductase with a concentration of 1 mg / mL and a pH of 7.0 Ferric ox...

experiment example 1

[0045] Take 200 μL of the agarose composite microspheres of Example 1 (the total gel mass is 0.3 mg, and the number of microspheres is 2.0×10 5 ) into a glucose solution with a concentration of 10 mmol / L, reacted for 30 min, added Griess reagent (nitric oxide detection reagent), put it into an ultraviolet spectrophotometer to detect the characteristic absorption peak, and unloaded nitric oxide precursor Enzyme-coated agarose microspheres were used as blank control.

[0046] figure 2 It is the ultraviolet spectrogram of the agarose composite microsphere. It can be seen from the figure that the agarose composite microsphere produces nitric oxide, which makes the Griess reagent have a characteristic absorption peak at 540nm.

[0047] image 3 is the curve of nitric oxide production versus time. It can be seen from the figure that as time goes by, the content of nitric oxide increases in an S-curve.

[0048] from figure 2 with image 3 It can be known that the agarose comp...

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Abstract

The invention discloses a composite agarose microsphere and a preparation method and application thereof. The composite agarose microsphere comprises an enzyme A, an enzyme B, a nitric oxide precursorderivative and agarose, wherein the enzyme A, the enzyme B and the nitric oxide precursor derivative are wrapped in the agarose to form the composite agarose microsphere, the enzyme A is glucose oxidase or glucose-like oxidase, and the enzyme B is peroxidase or like-peroxidase. The preparation method comprises the following steps of: adding the enzyme A and the enzyme B into an agarose solution to form a water phase; adding the water phase into an organic phase dropwise, performing stirring and centrifuging, and collecting a centrifuged product; and adding the nitric oxide precursor derivative into the centrifuged product for incubation to obtain the composite agarose microsphere. The prepared composite agarose microsphere can stimulate a gel composite microsphere to generate nitric oxideunder controllable catalysis by taking endogenous glucose as a reaction substrate, and has the advantages of long drug effect time and good biological safety; the preparation method is simple in operation and convenient to popularize and apply; and the composite agarose microsphere is applicable to preparing medicaments for preventing and treating cardiovascular diseases.

Description

technical field [0001] The invention relates to the technical field of materials, in particular to composite agarose microspheres and a preparation method and application thereof. Background technique [0002] Nitric oxide (NO) is a new type of biological information molecule, which has the functions of dilating blood vessels, regulating blood pressure, and inhibiting platelet adhesion. Physiological and pathophysiological processes. Hybridization of the two enzymes and the gel system to construct gel microspheres coated with cascade enzymes can give full play to the advantages of the two to generate nitric oxide, which is expected to be applied in biomedicine, analytical catalysis and other fields. [0003] At present, supplementing nitric oxide and increasing blood nitric oxide concentration is an important measure for the prevention and treatment of cardiovascular diseases such as thrombosis and hypertension. The supply of nitric oxide is mainly dependent on the supplem...

Claims

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Application Information

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IPC IPC(8): A61K9/50A61K47/26A61K38/54A61P7/02A61P9/12B01J13/02A61K31/155
CPCA61K9/5015A61K31/155A61K38/54A61P7/02A61P9/12B01J13/02C12Y101/03004C12Y111/01007A61K2300/00
Inventor 刘剑波李海丰何晓晓王柯敏刘松杨
Owner HUNAN UNIV
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