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Novel reagent and method for treating malignant cerebral malaria

A brain type and preparation technology, applied in the intersection of immunology and biomedicine, nanotechnology, and bionics, can solve the problems of not being able to relieve symptoms in a timely manner, and having no targeted therapeutic effect

Active Publication Date: 2020-10-30
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the therapeutic mechanism of artesunate is to kill the plasmodium in red blood cells, and it has no targeted therapeutic effect on the accumulation of inflammatory cells and cerebral edema in cerebral malaria, so it cannot relieve symptoms in time

Method used

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  • Novel reagent and method for treating malignant cerebral malaria
  • Novel reagent and method for treating malignant cerebral malaria
  • Novel reagent and method for treating malignant cerebral malaria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1. Preparation of ferritin nanozyme

[0049] Natural human ferritin is a globular iron storage protein composed of 24 heavy chain or light chain subunits in any proportion. The heavy chain and light chain subunits have high homology, and the molecular weights are 21kDa and 19kDa respectively (Theil EC., (1987) Annu. Rev. Biochem., 56:289-315). The subunit composition of ferritin in different organs and tissues of the human body is different. The heavy chain subunit is the main component in the heart, while the light chain subunit is more in the liver. Flexible supply of iron ions. However, only the heavy chain subunit can use oxygen to convert ferrous ions into ferric ions, so that iron ions can enter ferritin smoothly, so increasing the heavy chain subunit can increase the ability of the cell to utilize iron; while the light chain Subsequent to yacidol, it can increase the efficiency of iron storage. In the spherical shell of ferritin, iron ions form crys...

Embodiment 2

[0054] Example 2. Catalase activity of ferritin nanozymes

[0055] Based on the report of Fan Chunhai's research group, magnetic iron oxide nanoparticles have catalytic activity similar to catalase [12] , since ferritin is wrapped with an iron oxide inner core, it should have catalase-like activity, which can decompose hydrogen peroxide to generate oxygen. We used hydrogen peroxide to detect the enzymatic activity of ferritin nanozyme as follows: add 0.5mM H to ferritin nanozyme 2 o 2 React in PBS (pH=7.4) at 37° C. for 5 minutes, and observe the generation of bubbles. Such as figure 1 B, when ferritin nanozyme (Fenozyme) and H were added into the EP tube 2 o 2 After that, a large number of bubbles were generated in the liquid, but the ferritin shell (HFn) did not decompose hydrogen peroxide, indicating that the ferritin nanozyme had catalase-like activity.

Embodiment 3

[0056] Example 3. Ferritin Nanozyme Binding to Brain Microvascular Endothelial Cells

[0057] In order to study the binding of ferritin nanozyme to mouse microvascular endothelial cells, paraffin sections of mouse brain tissue and mouse brain microvascular endothelial cell line bEnd.3 were selected for incubation with fluorescent molecularly labeled ferritin nanozyme, and laser confocal microscopy was used and flow cytometry to detect the binding of ferritin nanozyme to brain microvascular endothelial cells.

[0058] The experimental method is as follows: according to the labeling method provided in the instructions, NHS-activated FITC (FITC-NHS, purchased from GE Healthcare) was labeled on ferritin nanozyme. Dewaxing according to the immunofluorescence steps of paraffin sections: take out the slices, and put them into xylene solution twice, 10 minutes each time; Hydration in neutral, 3 minutes per step; 0.01M (pH6.0) citric acid buffer, 100°C water bath for 30 minutes to rep...

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Abstract

The invention provides a novel reagent applicable to treatment of malignant cerebral malaria. The reagent is composed of a cavity protein shell formed by self-assembling of 24 protein subunits and aniron-based nano-enzyme with catalase activity, and can specifically target brain microvascular endothelial cells and remove ROS. Since an iron ion channel is formed in the protein shell of the reagent, nanometer iron core with uniform particle size can be synthesized in a cavity of the protein shell, and the nanometer iron cores have catalase catalytic activity and can catalyze decomposition of hydrogen peroxide; and the ferritin shell can target cerebral endothelial cells, promote proliferation of macrophages in the liver and polarization of the macrophages to the M1 subtype, and enhance thephagocytic function of the macrophages on infected red blood cells. Therefore, the reagent can be used for treating cerebral malaria.

Description

technical field [0001] The invention belongs to the intersecting fields of nanotechnology, bionics, immunology and biomedicine. In particular, the present invention provides a method for treating cerebral malaria that integrates specific recognition of cerebral malaria, symptom relief and promotion of immune function of the body. Background technique [0002] Malaria is currently the most serious tropical infectious disease threatening human health. According to World Health Organization estimates, more than 200 million people were living with malaria in 2017, and more than 400,000 of them died from malaria [1] . Among the five species of malaria parasites that currently host humans, Plasmodium falciparum is the most important threat. Cerebral malaria caused by Plasmodium falciparum infection is a form with a high mortality rate. Although the current artemisinin combination therapy based on the antimalarial drug artemisinin has achieved good results in the treatment of m...

Claims

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Application Information

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IPC IPC(8): A61K9/51A61K47/42A61K33/26A61P33/06G01N21/64
CPCA61K9/5169A61K33/26A61P33/06G01N21/6486Y02A50/30
Inventor 阎锡蕴范克龙赵帅
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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