New application of lycorine serving as HMGB1 protein inhibitor

A kind of protein inhibitor, technology of lycorine, applied in the field of lycorine compounds

Active Publication Date: 2015-12-30
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are currently no studies showing that lycorine

Method used

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  • New application of lycorine serving as HMGB1 protein inhibitor
  • New application of lycorine serving as HMGB1 protein inhibitor
  • New application of lycorine serving as HMGB1 protein inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Sensitivity experiment of normal cells and MM cells

[0030] In this example, normal human immortalized B lymphocytes and MM cells were treated with the same concentration of lycorine, and the cell survival status was detected by the CCK-8 method to evaluate the sensitivity of different cell types to lycorine. The experimental steps are as follows:

[0031] 1. Collect ARH-77, ANBL-6, NCI-H929, MM.1S cells in the logarithmic growth phase and normal human immortalized B lymphocytes, adjust the concentration of the cell suspension, add 100 μL to each well, and plate on a 96-well plate to make The cell density to be tested is 2×10 5 a / mL;

[0032] 2. Set up a zero-adjustment group without adding cells and only medium, and a DMSO control group without adding drugs at an appropriate concentration, and set 3-5 parallel wells for each concentration;

[0033] 3. Add 2.5μM, 5M, and 10μM concentrations of lycorine respectively, and place in 5% CO 2 , incubate in a ...

Embodiment 2

[0036] Example 2: Lycorine inhibits autophagy experiment

[0037] In this example, ARH-77, ANBL-6, NCI-H929 and MM.1S cells were treated with different concentrations of lycorine to inhibit autophagy. Then use the WesternBlot method, transmission electron microscopy and the method of exogenously introducing the pcDNA3.1-LC3-GFP plasmid to detect the inhibitory ability. The experimental steps are as follows:

[0038] 1. After ARH-77, ANBL-6, NCI-H929 and MM.1S cells were treated with different concentrations of lycorine, the changes of the key autophagy protein LC3B were detected by Western Blot method;

[0039] ① Collect ARH-77, ANBL-6, NCI-H929 and MM.1S cells in the logarithmic growth phase, and adjust the concentration of the cell suspension to 5×10 5 cells / mL in a cell culture flask with a total volume of 10 mL, then add lycorine at a concentration of 2.5 μM, 5 μM, and 10 μM respectively, and use the corresponding concentration of DMSO treatment group as a control, and ...

Embodiment 3

[0056] Example 3: Experiment of lycorine down-regulating HMGB1 protein level

[0057] In this example, the proteomics method was used to detect the down-regulation of HMGB1 protein in ARH-77 cells after different concentrations of lycorine were treated, and then the Western Blot method verified the HMGB1 protein of ARH-77 and ANBL-6 cells after different concentrations of lycorine. Both were down-regulated and showed a gradient change. At the same time, the expression of HMGB1 protein in normal human immortalized B lymphocytes, ARH-77 and ANBL-6 cells was detected by Western Blot method, and the experimental steps were as follows:

[0058] 1. The down-regulation of HMGB1 protein in ARH-77 cells was detected by proteomics method after the treatment of lycorine;

[0059] ① Collect ARH-77 cells in the logarithmic growth phase and adjust the concentration of the cell suspension to 5×10 5 cells / mL in a cell culture flask with a total volume of 10 mL, then add lycorine at a conc...

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Abstract

The invention discloses a new application of lycorine serving as HMGB1 protein inhibitor. Lycorine comprises lycorine or hydrates thereof or salt capable of being accepted pharmaceutically. It is proved through a series of experiments that lycorine has the high inhibiting effect on HMGB1 protein and can serve as the HMGB1 protein inhibitor. Diseases such as multiple myeloma caused by HMGB1 protein expression abnormality can be treated by means of lowering the HMGB1 protein level, and lycorine is expected to be developed into novel anti-tumor medicine.

Description

technical field [0001] The invention relates to the technical field of lycorine compounds, in particular to the new application of lycorine as an HMGB1 protein inhibitor and a drug for treating multiple myeloma. Background technique [0002] Human HMGB1 consists of 215 amino acid residues and has a molecular weight of about 25KD. It is named for its high mobility in polyacrylamide gel electrophoresis. HMGB1 is a high-abundance chromosome-binding protein in cells, and its functions are relatively complex. In addition to participating in the construction of nucleosomes, DNA replication, recombination, repair and gene transcription, it is also involved in the pathogenesis of various chronic inflammation and autoimmune diseases. High levels of serum HMGB1 expression can be detected in most inflammatory diseases, such as sepsis, rheumatoid arthritis, systemic lupus erythematosus, atherosclerosis, chronic kidney disease, and tumor development, invasion, and metastasis. [0003] A...

Claims

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Application Information

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IPC IPC(8): A61K31/4745A61P35/00
Inventor 刘静萧小鹃胡维新罗伊梁龙罗誉皓周卫华
Owner CENT SOUTH UNIV
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