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Application of TRAF6 inhibitor as and/or in preparation of ferroptosis inducer

A technology of inhibitors and inducers, applied in the field of application of TRAF6 inhibitors as and/or in the preparation of ferroptosis inducers, capable of solving problems such as unreported

Active Publication Date: 2021-11-09
GUANGDONG ZHANJIANG PROVINCIAL LAB OF SOUTHERN MARINE SCI & ENG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the application of oridonin as a TRAF6 inhibitor in the preparation of ferroptosis inducers has not been reported

Method used

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  • Application of TRAF6 inhibitor as and/or in preparation of ferroptosis inducer
  • Application of TRAF6 inhibitor as and/or in preparation of ferroptosis inducer
  • Application of TRAF6 inhibitor as and/or in preparation of ferroptosis inducer

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Experimental program
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preparation example Construction

[0133] The preparation method of the reagent or reagent in this embodiment is as follows:

[0134] Winter Lingnin Storage Hydram: Winter Trancin powder is placed in a sterile centrifugal tube with electronics, then adding dimethyl sulfoxide (DMSO), formulated a concentration of 20 mm.

Embodiment 1

[0135] Example 1 TRAF6 inhibitor induced iron death

[0136] (1) Cell transfection: H358 cells with 8 × 10 4 The density of / ml is inoculated in a 12-well plate, and the cells were cultured to 70% to 80% cell fusion per well for transfection: TRAF6-siRNA or NC-siRNA was carried out using Lipofectamine 2000 (Invitrogen, USA). Transfection (negative control NC-siRNA and Traf6-siRNA) were purchased in Sangonbiotech (Shanghai, China), of which traf6-siRNA sequence is TggattctCactGGCAAA, SEQ ID NO: 1, SEQ ID NO: 1 in t equivalent to U), NC- The final concentration of siRNA and TRAF6-siRNA was 10 nm; finally analyzed the expression level of the target protein in a cell transfected 48h, and the successful transfected cells were selected for subsequent experiments.

[0137] (2) Western Blot Detection TRAF6, C-MYC, Cyclin D1, GPX4, SLC7A11, FTH1 expression: cell protein is successfully transfected with RIPA cleavage liquid, collecting cellular protein; collected protein Samples were subje...

Embodiment 2

[0141] Example 2 Winter Ling Graphicine oridonin as Traf6 inhibitor induced iron death

[0142] (1) PD-L1 fluorescent antibody detection intracellular PD-L1 content: H358 cells in 1 × 10 5 / ml density in a 12-well plate, per well volume is 1 mL, and in 5% CO 2 The 37 ° C incubator was cultured overnight, and the second day was processed: 1) Divided into a blank control group (CONTROL) and the Winter Lingnin group (the final concentration of 2.5, 5, 10 μm respectively); and 2) Divided into blank control group (Control), gamma interferon (IFN-γ, final concentration of 20 ng / ml) group and IFN-γ (final concentration of 20 ng / ml) + Winter Lingnin (Oridonin, final concentration) The 10 μm group was administered for administration, and each treatment was treated with three complex apertures, and the cells were incubated with the PD-L1 fluorescent antibody (BioLegend, 393606) in a fridge in the 4 degree refrigerator; finally purified with PBS Cells, resuspended cells After detecting ...

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PUM

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Abstract

The invention belongs to the technical field of medicines, and discloses application of a TRAF6 inhibitor as and / or in preparation of a ferroptosis inducer. The invention discloses the application of the TRAF6 inhibitor as and / or in preparation of the ferroptosis inducer for the first time on the basis of a fact that an inventor finds that the TRAF6 inhibitor can induce iron accumulation, induce accumulation of intracellular active oxygen, lipid active oxygen and mitochondrial active oxygen and induce mitochondrial damage, and also can inhibit expression of C-Myc, Cyclin D1, GPX4, SLC7A11 and FTH1; and therefore, the TRAF6 inhibitor can be used as the ferroptosis inducer and can be used for preventing and / or treating tumors.

Description

Technical field [0001] The present invention belongs to the technical field of the drug, and more particularly to the application of TRAF6 inhibitors in and / or preparation of iron diential inducers. Background technique [0002] In 2012, it was found that an iron-dependent cell death form was discovered when studying Erastin killing Ras mutations - iron death, mainly in the cell "iron" alkindexic oxygen free radicals increased, redox steady state Unbalanced. Iron death has high metabolism and the characteristics of multi-target, opening up a new field of targeted therapy research. The main feature of iron death is the aggregation of ferrous ion accumulation and peroxide. The intracellular free iron reacts with hydrogen peroxide, catalyzed hydroxyl radicals, resulting in a component of the phospholipid film - a polydisaturated fatty acid peroxidation, causing the membrane unstable or even rupture, and ultimately leads to the process of cell death. Classic iron death regulation d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00
CPCA61K45/00A61K31/7088A61K31/352A61P11/00A61P35/00A61P43/00Y02A50/30
Inventor 罗连响黄芳芳许广香李晓玲罗辉
Owner GUANGDONG ZHANJIANG PROVINCIAL LAB OF SOUTHERN MARINE SCI & ENG
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