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New application for lycorine and bortezomib composition medicine

A technology of bortezomib and lycorine, which is applied in the field of new pharmaceutical uses of lycorine and bortezomib compositions, can solve problems such as drug resistance, and achieves improved sensitivity, improved cell proliferation, and inhibited cell proliferation. Effect

Active Publication Date: 2015-11-25
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the widespread use of bortezomib, myeloma cells are resistant to bortezomib clinically.

Method used

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  • New application for lycorine and bortezomib composition medicine
  • New application for lycorine and bortezomib composition medicine
  • New application for lycorine and bortezomib composition medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: In this example, the CCK-8 method was used to detect the effects of lycorine hydrochloride and bortezomib on the survival rate of MM cells after single and combined use.

[0020] Experimental steps:

[0021] (1) Collect ARH-77 and ANBL-6 cells in the logarithmic growth phase, 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 2×10 5 a / mL;

[0022] (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;

[0023] (3) Add 10 μM lycorine hydrochloride, 15 nM bortezomib, and 10 μM lycorine hydrochloride and 15 nM bortezomib respectively, and place in 5% CO 2 , incubate in a 37°C cell culture incubator for 24 hours;

[0024] (4) Add 10 μL CCK-8 solution to each well, incubate at 37°C in the dark for 2 hours, measure the OD valu...

Embodiment 2

[0026] Example 2: In this example, the CCK-8 method was used to detect the synergistic effect on MM cells after combined use of different concentration ratios of lycorine hydrochloride and bortezomib on MM cells.

[0027] Experimental steps:

[0028] (1) Collect ARH-77 and ANBL-6 cells in the logarithmic growth phase, 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 2×10 5 a / mL;

[0029] (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;

[0030] (3) Add different concentration ratios of lycorine hydrochloride and bortezomib (2.5μM+5nM, 2.5μM+10nM, 2.5μM+15nM, 5μM+5nM, 5μM+10nM, 5μM+15nM, 10μM+5nM , 10μM+10nM or 10μM+15nM), placed in 5% CO 2 , incubate in a 37°C cell culture incubator for 24 hours;

[0031] (4) Add 10 μL CCK-8 solu...

Embodiment 3

[0033] Example 3: In this example, the Western Blot method was used to detect the effects of lycorine hydrochloride and bortezomib on ARH-77 and ANBL-6 cells alone and in combination on autophagy-related proteins LC3B and Beclin1.

[0034] Experimental steps:

[0035] (1) Collect ARH-77 and ANBL-6 cells in logarithmic growth phase, adjust the concentration of cell suspension to 5×10 5 cells / mL in a cell culture flask with a total volume of 10 mL, and then add 10 μM lycorine hydrochloride, 15 nM bortezomib, and 10 μM lycorine hydrochloride and 15 nM bortezomib in combination, and use the corresponding concentration of DMSO as a control group. Place in a cell culture incubator and incubate for 24 hours;

[0036] (2) Collect the cells, wash them twice with 5 mL of PBS, add an appropriate volume of RIPA cell lysate (containing 0.5% protease inhibitor cocktail) to each tube, mix well, lyse on ice for 30 minutes, sonicate on ice for 30 seconds, and centrifuge at 4°C After 15 min...

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Abstract

The invention provides new application for a lycorine and bortezomib composition medicine. The lycorine and bortezomib composition is used for preparing a medicinal preparation for treating multiple myeloma. Compared with the fact that lycorine and bortezomib are used separately, combined use of lycorine and bortezomib can realize a better cell proliferation inhibition effect, and cell autophagy can be inhibited through combined use. The composition greatly improves sensibility of multiple myeloma to bortezomib and is applicable to drug-resistant multiple myeloma which is difficult to treat.

Description

technical field [0001] The present invention relates to the technical field of the composition of lycorine and bortezomib, and more specifically relates to a new drug application of the composition of lycorine and bortezomib. Background technique [0002] Multiple myeloma (multiple myeloma, MM) is a malignant tumor derived from terminally differentiated B lymphocytes. It is characterized by the clonal proliferation of a large number of plasma cells in the bone marrow. MM is still incurable. The bone marrow microenvironment is closely related to the proliferation, migration, survival, drug resistance and bone disease of MM. The nuclear factor NF-κB signaling pathway is a transcriptional regulator that plays an important role in the pathogenesis of MM. Bone marrow stromal cells HS-5 cells can produce GM-CSF, M-CSF, LIF and other cytokines to promote the proliferation and drug resistance of MM, so HS-5 cells can well simulate the bone marrow microenvironment of MM in vitro. ...

Claims

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Application Information

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IPC IPC(8): A61K38/05A61K31/69A61P35/00A61K31/4745
Inventor 刘静萧小鹃周文梁龙罗伊陈慧勇
Owner CENT SOUTH UNIV
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