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Screening method of active component in multiple target point anti-tumor traditional Chinese medicine and application thereof

An active component and multi-target technology, which is applied in antineoplastic drugs, separation methods, medical preparations containing active ingredients, etc., can solve problems such as low efficiency, unsatisfactory results, and long time-consuming

Active Publication Date: 2015-07-29
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on anti-tumor mechanism has always been a hot spot and difficult point. Usually, we can only start with a certain protein or a certain biomacromolecule, which is like finding a needle in a haystack, which takes a long time, is inefficient and often results in unsatisfactory results.

Method used

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  • Screening method of active component in multiple target point anti-tumor traditional Chinese medicine and application thereof
  • Screening method of active component in multiple target point anti-tumor traditional Chinese medicine and application thereof
  • Screening method of active component in multiple target point anti-tumor traditional Chinese medicine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Preparation of silica gel column for screening of natural active components of multi-target anti-tumor traditional Chinese medicine

[0035] (1) Culture and expansion of MCF-7 cells:

[0036] MCF-7 cells were cultured and expanded with DMEM medium containing 10% calf serum (NBS), and the cell number concentration was adjusted to 1×10 7 pcs / ml, spare;

[0037] (2) Extraction of multi-target receptor enrichment region:

[0038] Add the lysate containing detergent to the MCF-7 cells collected in (1), and extract the multi-target receptor enrichment area; then use the sucrose density gradient centrifugation method to separate the receptor enrichment area, and the sucrose-containing The suspension of the receptor-enriched region was placed in 80 mM Tris-HCL solution with pH 7.2 at 4°C, dialyzed for 24 hours, and the solution containing the multi-target receptor-enriched region was placed in a -80°C refrigerator for later use.

[0039] (3) Preparation of silica ...

Embodiment 2

[0041] Example 2: Preparation of a chromatographic column for screening of natural active components of multi-target anti-tumor traditional Chinese medicine

[0042] (1) Culture and expansion of MCF-7 cells:

[0043] MCF-7 cells were cultured and expanded in DMEM medium containing 10% calf serum (NBS), and the cell number concentration was adjusted to 10×10 7 pcs / ml, spare;

[0044] (2) Extraction of multi-target receptor enrichment region:

[0045] Add the lysate containing detergent to the MCF-7 cells collected in (1), and extract the multi-target receptor enrichment area; then use the sucrose density gradient centrifugation method to separate the receptor enrichment area, and the sucrose-containing The suspension of the receptor-enriched region was placed in 120 mM Tris-HCL solution with pH 7.5 at 4°C, dialyzed for 20 hours, and the solution containing the multi-target receptor-enriched region was placed in a -80°C refrigerator for later use.

[0046] (3) Preparation of ...

Embodiment 3

[0048] Example 3: Identification of multi-target receptor modified silica gel stationary phase

[0049] Take the suspension of the silica gel stationary phase prepared above for the screening of natural active components of multi-target anti-tumor traditional Chinese medicine, add 200 μL of the primary antibody dilution (1:100) of EGFR, TNFR and Fas, and different fluorescent secondary antibodies respectively. Anti-dilution solution (1:500), observe the results under an inverted fluorescence microscope. The results observed under the fluorescence microscope were figure 2 , all the surface-specific fluorescence of silica gel stationary phase microspheres modified by multi-target receptors is uniform and bright. The experimental results showed that EGFR, Fas and TNFR were successfully connected on the silica gel stationary phase.

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Abstract

A method for screening natural active component in multiple target point anti-tumor traditional Chinese medicine, especially comprises: extracting a receptor enrichment region containing EGFR, TNFR, Fas from a human breast cancer MCF-7 cell strain, connected to a silica gel stantionary phase to form a separated silica gel column or a chromatographic column, realizing highly efficient screening of the active component in the multiple target anti-tumor traditional Chinese medicine, and changing the current situation of single target screening of the traditional Chinese medicine active component. Not only the screening method provides example for highly efficinet screening of the EGFR, TNFR and FAS inhibitors, but also the design thought provides mirror and reference value for the active component screening of other anti-tumor medicine, and it is ensured that the design thought has wide application space in the active component screening of the traditional Chinese medicine. The invention discloses a preparation method.

Description

technical field [0001] The invention relates to a normal-pressure separation column or chromatographic column for screening active components of multi-target anti-tumor traditional Chinese medicines. Background technique [0002] The research and development model of single-component and single-target drugs has been challenged more and more. The abnormal phenomenon that the cost of innovative drug development continues to rise, while the research and development efficiency is gradually decreasing, has aroused deep thinking among scholars. "Single-target, high-efficiency The drug research and development concept of affinity and high selectivity has been questioned more and more [see: Darras FH, Pockes S, Huang G, Wehle S, Strasser A, Wittmann HJ, et al. Synthesis, biological evaluation, and computational studies of Tri-and tetracyclic nitrogen-bridgehead compounds as potent dual-acting AChE inhibitors and hH3 receptor antagonists. ACS ChemNeurosci. 2014; 5:225-42; Darras FH, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/22G01N33/533A61K36/31A61P35/00
CPCG01N30/02G01N30/60
Inventor 徐希明王启龙余江南丁丽霞曹霞屈睿童姗姗
Owner JIANGSU UNIV
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