Use of mitochondrial complex I reversible inhibitor combined with active oxygen scavenger to prepare ischemia-reperfusion injury protection drug

A mitochondrial complex, ischemia-reperfusion technology, applied in the field of medicine, to achieve the effect of improving the therapeutic effect and improving the therapeutic effect

Active Publication Date: 2019-10-11
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no report on the reversible inhibitor of mitochondrial complex I used as an ischemic preconditioning drug, let alone the report on its combination with a reactive oxygen species scavenger for the protection of ischemia-reperfusion injury

Method used

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  • Use of mitochondrial complex I reversible inhibitor combined with active oxygen scavenger to prepare ischemia-reperfusion injury protection drug
  • Use of mitochondrial complex I reversible inhibitor combined with active oxygen scavenger to prepare ischemia-reperfusion injury protection drug
  • Use of mitochondrial complex I reversible inhibitor combined with active oxygen scavenger to prepare ischemia-reperfusion injury protection drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Therapeutic effect on myocardial ischemia-reperfusion

[0031] 1. Experimental materials

[0032] H9c2 cells, N2a cells, HK-2 cells: purchased from ATCC Company and preserved by our laboratory. DMEM high-glucose medium, fetal bovine serum, 0.25% trypsin, 100× penicillin and streptomycin, CalceinAM (Thermo Fisher, USA), PBS (Beijing Boaosen Biotechnology Co., Ltd.), CoCl 2 , hydrogen peroxide (US Sigma Company), PhosSTOP, completeULTRATablets, Mini EASYpack (Roche Company), CCK8, Fluo-4AM, Rhod-2AM (Shanghai Dongren Chemical Technology Co., Ltd.), RIPA lysate, LY294002 (Shanghai Biyuntian Bio Technology Co., Ltd.), AKT, p-AKT, p-GSK3, β-actin (Cell Signaling Technology, USA). Ultra-clean bench (Thermo Scientific 1300A2, Thermo, USA), multi-functional microplate reader FLUOstar Omega (BMG LABTECH, Germany), confocal laser (Carl Zeiss, Germany), electric constant temperature water bath (Guohua Electric Co., Ltd., HH- 4 digital display constant temperature wat...

Embodiment 2

[0101] Embodiment 2: Therapeutic effect on cerebral ischemia-reperfusion

[0102] 1. Experimental method

[0103] 1. Nerve cell culture and passage

[0104] (1) Nerve cell recovery

[0105] Take out the nerve cells (N2a cells) frozen in the liquid nitrogen tank, thaw them quickly in a water bath preheated to 37 ° C, and transfer the cells to 9 ml of DMEM medium containing 10% serum under aseptic conditions for centrifugation centrifuge at 1000rmp for 4min. Discard the supernatant, resuspend the cells in 1mL of the above medium, and then transfer to a culture dish containing new medium, gently blow the cells with a pipette gun to make them evenly distributed in the culture dish, and place at 37°C , 5%CO 2 Conditioned culture, the medium was changed after 24h.

[0106] (2) Cell digestion

[0107] Discard the culture medium in the culture dish, wash once with PBS solution, add 1ml of trypsin, place the culture dish in the cell culture incubator for 1min, when the cells shri...

Embodiment 3

[0124] Embodiment 3: to the therapeutic effect of renal ischemia-reperfusion

[0125] 1. Experimental method

[0126] 1. Kidney epithelial cell culture and passage

[0127] (1) Recovery of renal epithelial cells

[0128] Take out the renal epithelial cells (HK2 cells) frozen in the liquid nitrogen tank, thaw them quickly in a water bath preheated to 37°C, and transfer the cells to 9ml of DMEM medium containing 10% serum under aseptic conditions. In a centrifuge tube, centrifuge at 1000rmp for 4min. Discard the supernatant, resuspend the cells in 1mL of the above medium, and then transfer to a culture dish containing new medium, gently blow the cells with a pipette gun to make them evenly distributed in the culture dish, and place at 37°C , 5%CO 2 Conditioned culture, the medium was changed after 24h.

[0129] (2) Cell digestion

[0130] Discard the culture medium in the culture dish, wash once with PBS solution, add 1ml of trypsin, place the culture dish in the cell cult...

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Abstract

The invention discloses an application of Application of protective medicine for ischemia reperfusion injury prepared by mitochondrial complex I reversibility inhibitor and active oxygen scavenger. According to the invention, the mitochondrial complex I reversibility inhibitor can play a protective role similar to ischemic preconditioning, and can be used as an ischemic preconditioning medicament;the mitochondrial complex I reversibility inhibitor is combined with active oxygen scavenger, the mitochondrial complex I is administered reversibility inhibitor before ischemia, and is administeredactive oxygen scavenger before reperfusion, and the therapeutic effect of ischemia reperfusion can be obviously improved. According to the invention, the double-layer tablet is taken before ischemia,the outer quick release layer containing the mitochondrial complex I reversibility inhibitor quickly releases and play the role of ischemic preconditioning, the inner slow release layer containing reactive oxygen scavenger slowly releases and reaches the maximum plasma concentration before reperfusion, so as to improve the curative effect of ischemia reperfusion, and the medicine is effective andconvenient.

Description

technical field [0001] The invention belongs to the field of medicine, and specifically relates to the application of a reversible inhibitor of mitochondrial complex I and an active oxygen scavenger to prepare ischemia-reperfusion injury protection medicine. Background technique [0002] At present, myocardial ischemia has become a buzzword in the mouth of the people, the main reason is that the incidence of myocardial ischemia has been increasing in recent years, and the mortality rate has also increased significantly. In recent years, with the improvement of clinical diagnosis and treatment, the proportion of patients receiving blood reperfusion therapy after myocardial ischemia has increased. But sometimes myocardial ischemia-reperfusion often leads to a series of cardiac adverse events, which partially offset the beneficial effects on the heart, which is called reperfusion injury. It is characterized by apoptosis of cardiomyocytes, increased oxygen free radicals, increa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/06A61K45/00A61K9/24A61K31/58A61K31/155A61K31/355A61K31/198A61K31/11A61P9/10
Inventor 李萍郝海平杨华蒋黎锋曾昊倪丽红祁励丰荣
Owner CHINA PHARM UNIV
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