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Nano-enzyme diagnosis and treatment agent for acute kidney injury and preparation method and use of nano-enzyme diagnosis and treatment agent

An acute kidney injury and nanozyme technology, applied in the field of biomedical materials, can solve the problems of large toxic and side effects, hinder clinical application, low utilization rate of small molecule drugs, etc., and achieve good therapeutic effect, good water solubility and biological safety, The effect of excellent biocompatibility and biosafety

Pending Publication Date: 2021-03-19
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, small molecule drugs have low utilization, high toxicity and limited efficacy
These hinder their clinical application

Method used

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  • Nano-enzyme diagnosis and treatment agent for acute kidney injury and preparation method and use of nano-enzyme diagnosis and treatment agent
  • Nano-enzyme diagnosis and treatment agent for acute kidney injury and preparation method and use of nano-enzyme diagnosis and treatment agent
  • Nano-enzyme diagnosis and treatment agent for acute kidney injury and preparation method and use of nano-enzyme diagnosis and treatment agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1: synthetic nanozyme diagnosis and treatment agent

[0053] Nanozyme diagnostic agent synthesis: such as figure 1 As indicated, potassium ferricyanide (1 mmol / L, 20 ml) solution was added to 80 ml of chitosan solution (3 mg / ml), then stirred and added dropwise with ferrous chloride solution (1 mmol / liters, 20 ml). The reaction was stirred at 25°C for 1 hour. Then 120 mL of acetone was added. Centrifuge at 8000 rpm, wash the obtained blue precipitate several times with acetone, and dry to obtain the final product.

[0054] figure 1 is a roadmap for the synthesis of nanozyme diagnostic agents, where K 3 [Fe(CN)] 6 stands for potassium ferricyanide, CS stands for chitosan, FeCl 2 Represents ferrous chloride. The chitosan surface ligand in the nanozyme diagnosis and treatment agent can well stabilize the Prussian blue nanoparticles.

[0055] figure 2 is the AFM image of the synthesized nanozyme diagnostic agent; image 3 is the XRD pattern of the sy...

Embodiment 2

[0056] Example 2: The ability of nanozyme diagnostic agent to scavenge various active oxygen / active nitrogen and the ability of nanozyme diagnostic agent to scavenge hydroxyl radicals

[0057] The efficiency of scavenging hydroxyl radicals of different concentrations of nanozyme diagnostic agents (0-100 μg / mL) was determined by the hydroxyl radical antioxidant capacity (HORAC) kit (Cell Biolabs, Inc., USA). Testing was performed according to the protocol provided by the manufacturer.

[0058] Such as Figure 4 As shown, the nanozyme therapeutic agent can effectively scavenge hydroxyl radicals, and has a concentration-dependent characteristic.

[0059] The superoxide anion scavenging efficiency of different concentrations of nanozyme diagnostic agent (0-100 μg / mL) was determined by SOD detection kit (Sigma-Aldrich, USA). Testing was performed according to the protocol provided by the manufacturer.

[0060] Such as Figure 5 As shown, the nanozyme therapeutic agent can effec...

Embodiment 3

[0067] Example 3: Cytotoxicity of Nanozyme Therapeutic Agents and Protection of Kidney Cells by Removing Various Reactive Oxygen / Reactive Nitrogen The standard MTT method was used to evaluate the effect of nanozyme therapeutic agents on the survival rate of 293T renal embryonic cells.

[0068] 293T cells at 1×10 per well 4 Density seeded into 96-well plate, and placed in 37 degrees, 5% CO 2 Cultivate under the condition for 12h. Next, the old culture medium in the 96-well plate was sucked out, and culture medium solutions containing different concentrations of nanozyme therapeutic agent were added respectively. After continuing to cultivate for 20 hours, suck out the old medium in the 96-well plate, add 100 μL of MTT medium solution (0.8 mg / mL) to each well, and continue to cultivate for 4 hours. Aspirate the residual medium in the 96-well plate, add Add 150 μ L of DMSO solution to the solution, shake gently, detect the OD value of each hole on a Synergy H1 microplate reader...

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Abstract

The invention discloses a nano-enzyme diagnosis and treatment agent for acute kidney injury and a preparation method and use of nano-enzyme diagnosis and treatment agent. The nano-enzyme diagnosis andtreatment agent comprises Prussian blue nano-particles and surface ligands combined on surfaces of the Prussian blue nano-particles. The nano-enzyme diagnosis and treatment agent comprises the surface ligands (CS) and the Prussian blue nanoparticles (PB NZs) protected by the surface ligands. The nano-enzyme diagnosis and treatment agent has an ultra-small size, can be effectively enriched in thekidney of a mouse, can remove a large amount of active oxygen or active nitrogen in kidney tubules to relieve and treat acute kidney injury induced by glycerol or cis-platinum, and can be used as a contrast agent for nuclear magnetic resonance imaging and photoacoustic imaging technologies. In addition, the nano-enzyme diagnosis and treatment agent has a good treatment effect and also has excellent biocompatibility and biosafety.

Description

technical field [0001] The invention relates to the technical field of biomedical materials, in particular to a nanozyme diagnosis and treatment agent for acute kidney injury and its preparation method and application. Background technique [0002] Acute kidney injury is an important health problem in humans. Due to its high morbidity and mortality, it is estimated that 1.7 million people die every year worldwide. Currently, adjuvant therapy and kidney transplantation are the most common treatments. Recent studies have shown that the pathogenesis of acute kidney injury is associated with excess intracellular reactive oxygen species and reactive nitrogen species. Previously, some small-molecule drugs, such as amifostine and acetylcysteine, have been shown to act as antioxidants and eliminate reactive oxygen species to alleviate acute kidney injury. However, small molecule drugs have low utilization rate, high toxicity and limited efficacy. These hinder their clinical appl...

Claims

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Application Information

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IPC IPC(8): A61K49/22A61K49/18A61K49/12A61K9/51A61K47/36A61K33/26A61P13/12
CPCA61K49/225A61K49/1863A61K49/12A61K49/0002A61K9/5161A61K33/26A61P13/12
Inventor 黄鹏张东阳刘恒克林静
Owner SHENZHEN UNIV
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