Antitumor MA-TNF alpha medicine composition and application thereof

A composition and anti-tumor technology, applied in the field of anti-tumor MA-TNFα pharmaceutical composition, can solve the problems of weakening the anti-tumor effect of tumor necrosis factor, metastasis, tumor recurrence, etc., to achieve enhanced anti-tumor efficacy, reduce risk, The effect of reducing toxic side effects

Active Publication Date: 2013-02-20
BIORAY LABORATORIES INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The activation of NF-κB leads to the expression of a large number of anti-apoptotic genes, which greatly weakens the anti-tumor effect of tumor necrosis factor, and also leads to inflammation, causing tumor recurrence, metastasis and other side effects

Method used

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  • Antitumor MA-TNF alpha medicine composition and application thereof
  • Antitumor MA-TNF alpha medicine composition and application thereof
  • Antitumor MA-TNF alpha medicine composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1: cell proliferation experiment

[0044] Materials and Reagents:

[0045] Using Promega CellTiter AQueous One Solution Cell Proliferation Assay Kit. Microplate UV-visible continuous wavelength microplate reader cell line human pancreatic cancer cell line PANC-28, human breast cancer cell line MDA-MB-231 / 435, human colon cancer cell line HCT116, human liver cancer cell line HepG2, SK-HEP -1, blood lymphocyte K562. Human recombinant tumor necrosis factor α (Tumor Necrosis Factor α, TNF-α).

[0046] experimental method:

[0047] 1. Divide cells into 4.0×10 8 / L density inoculated in a 96-well culture plate, 100 μL per well, 37 ° C, 5% CO 2 .

[0048] 2. Culture in an incubator until the cells adhere to the wall, add different concentrations of maslinic acid (0, 0.5, 1, 1.5, 3, 5, 10, 25, 50 μM) for pretreatment for 6 hours, add 0.1nM TNF for 36 hours, ( Or pre-add 5 μM maslinic acid for 6 hours and then add different concentrations of TNF for 36 hours) ...

Embodiment 2

[0053] Example 2: Cell Infiltration Experiment

[0054] Experimental Materials:

[0055] Transwell small test was purchased from millipore company.

[0056] experimental method:

[0057] 1. Divide cells into 4.0×10 8 / L density inoculated in 10cm culture dish, 8ml / dish, 37℃, 5% CO 2 cultured in an incubator.

[0058] 2. When the cells grow to 80% full, replace with serum-free incomplete medium and continue starvation culture for 8 hours.

[0059] 3. Trypsinize, adjust the cells to 1×105 / ml cell suspension with incomplete medium without serum, then add 100ul cell suspension / well (control group and TNF group) on the upper layer of Transwell (control group and TNF group) or add hawthorn acid 100ul cell suspension / hole (maslinic acid group and maslinic acid plus TNF group), respectively add 600ul medium (control) or 600ul medium containing 0.2nM TNF to the lower layer of Transwell, after 2~4h, count migration to The number of cells on the lower membrane surface of the Transw...

Embodiment 3

[0063] Example 3: Live / Dead assay

[0064] Reagents and methods:

[0065] Reagent materials Viability / Cytotoxicity Assay Kit for Animal Live & Dead Cells (Biotium, Inc.30002)

[0066] Experimental steps, according to the operating steps of the kit instructions.

[0067] 1. Divide cells at 4.0 x 10 8 Inoculate in a 96-well culture plate at a density of / L, 100 μL per well, culture in a 37°C, 5% CO2 incubator until the cells adhere to the wall, add corresponding drugs, and repeat 3 wells for each concentration.

[0068] 2. Cultivate in a 37°C, 5% CO2 incubator for 24-36 hours.

[0069] 3. Wash the cells once with PBS, add 1 ulethidium homodimer and calcein-AM respectively, let stand at room temperature for 30 minutes, observe and count the cells under a fluorescence microscope, red is dead cells, green is live cells.

[0070] 4. Calculate the cell death rate according to the following formula: cell death rate=number of dead cells / total number of cells×100%.

[0071] Experim...

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Abstract

The invention relates to an antitumor medicine composition and application thereof. The composition comprises maslinic acid or physiologically acceptable salts or derivatives thereof and tumor necrosis factor alpha used in combination. The acting effects of the composition are superior to the added effects of each medicine used individually. The combination use can effectively reduce the toxic and side effects produced by the tumor necrosis factor alpha used individually, so that the composition used in combination can be used for treating tumor and related diseases.

Description

technical field [0001] The present invention relates to an anti-tumor pharmaceutical composition and its application, in particular to an anti-tumor MA-TNFα pharmaceutical composition and its application. Background technique [0002] The incidence of cancer is still increasing year by year, and cancer is currently the number one cause of death. Although, intensive and extensive research and investigation in recent years have provided a relatively in-depth understanding of the mechanism of tumor generation. In theory, it clarifies the various processes that can control the development of tumors by various means. In practice, many different anti-tumor drugs that act on different stages of cancer through different mechanisms have been discovered and accumulated. Especially in the past three decades, many anti-tumor drugs have entered clinical practice one after another. However, tumor is a complex disease caused by many factors. Practice has proved that the therapeutic eff...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/19A61P35/00A61K31/56
Inventor 刘明耀罗剑李成海杨正峰仇文卫汤杰
Owner BIORAY LABORATORIES INC
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