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Methotrexate and berberine compound pharmaceutical composition and application thereof

A technology of methotrexate and berberine, applied in the field of natural medicine, to achieve the effect of reducing toxic and side effects, strong anti-proliferation activity, and good prospects for clinical drug development

Inactive Publication Date: 2018-12-07
FOSHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Combination of methotrexate and berberine for the prevention and treatment of malignant tumor lymphoma has not been reported in the relevant literature almost no

Method used

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  • Methotrexate and berberine compound pharmaceutical composition and application thereof
  • Methotrexate and berberine compound pharmaceutical composition and application thereof
  • Methotrexate and berberine compound pharmaceutical composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Screening optimized methotrexate (MTX) and berberine (BER) compound pharmaceutical composition using Daudi cells, Raji cells and BC-3 cells

[0023] Take the cells in the logarithmic growth phase and inoculate 3 × 10 4 Cells / well were placed on a 96-well plate. After 6 hours of growth, the supernatant was discarded by centrifugation, and then administered in the following groups: Tumor cells were divided into a drug-free group and a drug-added group, and the drug-added group was divided into MTX and BER single drugs. Group, MTX and BER combined with different molar ratio groups, each group set 4-6 duplicate wells, cultured for 24 hours, discarded the supernatant, added 100 μl MTT (tetrazolium salt) serum-free medium containing 0.5 mg / ml for culture After 4 hours, 100 μl of DMSO (dimethyl sulfoxide) was added, placed on a micro-shaker for 10 min, and then placed on a microplate reader to detect the OD value at 570 nm. Results The inhibition rate of tumor cell...

Embodiment 2

[0030] Example 2: Analysis of the effect of combined administration of MTX and BER

[0031] Based on the Median-effect Principle (median-effect principle or Chou-Talalay combined index method), the CombiDrug statistical software was used to draw the dose-response curve and the combined exponential curve (fa-C curve) under different effects. The relationship between the effect and the combination index quantitatively evaluates whether the two drugs are synergistic, antagonistic or additive. Specific steps are as follows:

[0032] The drug effect is the inhibition rate (fa)=1-(the average OD570 value of the test group / the average OD570 of the tumor cell blank control group) according to the middle-effect formula fa / fu=(D / Dm) m , take the logarithm logfa / fu=mlogD–mlogDm on both sides, set a=-mlogDm, b=m, x=logD, y=logfa / fu, and substitute it into the middle-effect equation to get y=bx–a; where fa is the drug effect , fu=1-fa, D is the drug concentration, m is the slope, Dm is t...

Embodiment 3

[0037] Example 3: Safety of MTX and BER on peripheral blood mononuclear lymphocytes

[0038] The safety of MTX and berberine on PBMC cells was evaluated after isolation and culture of healthy volunteer monocyte lymphocyte PBMC ( figure 1 ): ①Draw fresh blood from healthy volunteers into a heparin anticoagulation tube (sterile); then resuspend the cells with an equal volume of PBS (or serum-free D-Hank buffer) (sterile); ②Resuspend the cells Add pre-laid human lymphocyte separation solution (preheated at 37°C), and the volume ratio of lymphocyte separation solution and cell suspension is not less than 1:1; ③ 500×g (or 2000rpm) horizontal centrifuge for 20-30min, room temperature (20-30℃) slow speed; ④ Discard the upper plasma, carefully aspirate the middle white mist layer and add it to 5ml (or 1-2 times the volume) of PBS (or serum-free cell culture buffer), 200×g or 1000rpm Centrifuge for 10 minutes at room temperature (20-30°C), at a slow speed, discard the supernatant, and...

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Abstract

The invention belongs to the field of natural medicines and discloses a methotrexate and berberine compound pharmaceutical composition and application thereof. Active ingredients of the compound pharmaceutical composition are composed of methotrexate and berberine at the molar ratio of 1: 1-1: 100. By combination of methotrexate and berberine, the drug combination has strong application in the collaborative treatment of lymphoma.

Description

technical field [0001] The invention belongs to the field of natural medicines, and particularly relates to a compound medicinal composition of methotrexate and berberine and uses thereof. Background technique [0002] Cancer is a major stubborn disease that seriously endangers human health, and it has become the second largest killer after cardiovascular disease. Since the beginning of the 21st century, due to the rapid process of modern industrialization, environmental pollution and many other factors, the immune system of lymphocytes has been destroyed and the incidence of lymphoma has been increasing year by year. The drug system appears very urgent. [0003] Methotrexate (Mthotrexate, MTX) is an anti-folate metabolism tumor drug. Similar in chemical structure to folic acid, dihydrofolate (FH) is produced by competitively inhibiting the activity of dihydrofolate reductase (DHPR) in cells 2 ) is reduced to tetrahydrofolate (FH 4 ) is hindered, resulting in insufficien...

Claims

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Application Information

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IPC IPC(8): A61K31/519A61K31/4375A61P35/00
CPCA61K31/519A61K31/4375A61P35/00A61K2300/00
Inventor 刘连李亚君刘腾彭咏波崔亮李雄刘武昆刘靖谢国健
Owner FOSHAN UNIVERSITY
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