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Application of flavonoid

A kind of technology of flavonoids, hydroxyflavonoid dihydrate

Inactive Publication Date: 2019-09-06
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these drugs are limited by factors such as lack of oral availability, limited tissue and tumor penetration, and high cost.

Method used

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  • Application of flavonoid
  • Application of flavonoid
  • Application of flavonoid

Examples

Experimental program
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Embodiment 1

[0029] (1) Screening of inhibitors of PD-1 / PD-L1 protein interaction

[0030] ① Dilute the His-Trx-PD-1 fusion protein expressed by E. coli to 1 μg / mL with carbonate coating buffer, add 100 μL per well to a 96-well microtiter plate, and place at 4 °C overnight. Wash 3 times with 1×PBST, block with 5% skimmed milk powder, 300 μL per well, block at 37°C for 2hrs.

[0031] ② Wash 3 times with 1×PBST. At the same time, dilute GST and GST-PD-L1 proteins to 2 μg / mL with 1×PBS, add 0.2 μL of DMSO or small molecule compound (final concentration 5 μM), mix well, add 100 μL to each well of a 96-well plate, and combine at 37 °C 2hrs. Wash 3 times with 1×PBST, add 100 μL of GST antibody (rabbit polyclonal antibody) diluted in 1×PBS to each well, and bind at 37°C for 2hrs. Wash 3 times with 1×PBST, add 100 μL of HRP-labeled goat anti-rabbit antibody diluted in 1×PBS to each well, bind at 37°C for 0.5hrs.

[0032] ③Wash 5 times with 1×PBST, add 100 μL TMB substrate chromogenic solution ...

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Abstract

The invention relates to an application of a flavonoid compound, the flavonoid compound is 3, 3', 4', 5, 7-pentahydroxyflavone dihydrate, the ELISA and protein fluorescence quenching experiments provethat the compound can effectively inhibit interaction of PD-1 / PD-L1 proteins, and has a strong affinity for PD-L1 protein. Cell clone number formation and ELISA detection of IL-2 secretion assay prove that the compound can effectively enhance the tumor killing activity of T lymphocyte Jurkat cells against MDA-MB-231 and NCI-H460 tumor cells as well as the expression level of IL-2, and then reverse T cellular immunosuppression. The research results of the application provide a drug for tumor immunotherapy targeting the PD-1 / PD-L1 immunological test site.

Description

technical field [0001] The invention relates to the application of a small molecular compound, especially the application of a flavonoid compound. Background technique [0002] T cells are important immune cells and play an important role in the process of anti-tumor immunotherapy. Activation of T cells is a highly regulated process that requires, in addition to the first signal transduced by the T cell receptor (TCR) (TCR binds to the MHC-antigen peptide), co-signaling molecules (co-stimulatory and co-inhibitory) ) to promote or inhibit T cell responses, wherein the second signal is the key to eliciting T cell antigen-specific responses. Co-signaling molecule receptor proteins include molecules such as CD28, PD-1, CTLA-4, ICOS, BTLA, LAG3, TIM3, and OX40, among which CD28, OX40, and ICOS are used to present co-stimulatory signals to activate T cell immune responses, PD-1, CTLA-4, BTLA, LAG3, and TIM3 receptor proteins are used to present co-inhibitory signals to suppress ...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61P35/00
CPCA61K31/352A61P35/00
Inventor 刁爱坡景磊李玉银
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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