Purpose of polyether compound in medicine preparation and anti-tumor medicine screening method

A technology for preparing drugs and compounds, which is applied in the field of biomedicine and can solve problems such as the use of polyether antibiotics to be studied

Inactive Publication Date: 2018-04-24
WUHAN J1 BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the use of polyether antibiotics in the preparation of drugs remains to be studied

Method used

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  • Purpose of polyether compound in medicine preparation and anti-tumor medicine screening method
  • Purpose of polyether compound in medicine preparation and anti-tumor medicine screening method
  • Purpose of polyether compound in medicine preparation and anti-tumor medicine screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The preparation method of embodiment 1 polyether compound

[0042] One Streptomyces nanchangensis subtype, two Streptomyces hygroscopicus subtypes and one Streptomyces endus subtype Streptomyces were inoculated on the SFM plate, and cultured for about three to four days. When colonies are formed in the SFM medium, pick a single colony and culture it in the seed medium for about three to four days, and inoculate it into the fermentation medium according to 1% amount (the composition of the fermentation medium is 30g / L of soluble starch, yellow Bean cake powder 10g / L, yeast extract 2.5g / L, CaCO 3 3g / L, pH 7.2). Culture conditions: 30 degrees, 220 rotation speed, culture for 7 to 8 days.

[0043] When the culture reaches the 7th or 8th day, collect the fermentation broth and centrifuge at a high speed to separate the supernatant from the mycelium, add an equal amount of acetone to the mycelium to destroy the bacteria, ultrasonicate for 20 minutes, spin evaporate acetone...

Embodiment 2

[0054] Example 2 Research on J1001-2 Sodium Salt by Proteomics and Phosphoproteomics

[0055] (1) Cell culture

[0056] MDA-MB-231 cells (triple-negative breast cancer cells) were cultured with DEME containing 10% fetal bovine serum, 80 μg / m L streptomycin, and 80 U / m L penicillin at 37°C and 5% CO 2 Routine cultivation in an incubator. When the cells are about 90% full, add the drug J1001-2 sodium salt. In the protein group experiment, 3 administration groups (administration concentration of 2 μM) were set up, respectively 24h, 48h, 72h and a blank group (no drug addition). In the phosphorylated protein group experiment, two administration groups (administration concentration of 3 μM) were set up for 6h, 12h and a blank group (no drug addition).

[0057] (2) Experimental methods of proteomics

[0058] 1) Proteome sample preparation

[0059] Collect the cells of the blank group and the administration group, transfer to a 1.5mL centrifuge tube, centrifuge at 2500rpm at 4°C...

Embodiment 3

[0080] Example 3 Study on the Targets of Polyether Compounds for Inhibiting Tumors

[0081] 1. Experimental method—western blotting experiment

[0082] Protein gel concentration: 8% precast gel, 10% precast gel, 12% precast gel, 4-20% precast gel (from GenScript)

[0083] Set up the administration group as shown in the figure below, collect cells 16 hours after administration, digest with 0.125% trypsin, wash twice with PBS, centrifuge, add 400L of RIPA per 10cm dish cell volume (in which protease inhibitors and phosphorylation are added to RIPA Protease inhibitors) lyse the cells on ice for 1-2 hours, and centrifuge at 4 degrees at high speed (12000rmp×10min). Aspirate the supernatant as the protein.

[0084] Protein loading amount: (uniform loading amount 25ng) (MDA-MB-231 cells)

[0085] BCA determination of protein concentration standard curve such as Image 6 shown.

[0086] Calculate the concentration (in μg / mL) through the standard curve as follows:

[0087] ...

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Abstract

The invention discloses a purpose of a polyether compound in medicine preparation and an anti-tumor medicine screening method. The medicine is used for regulating and controlling at least one of the following signal channels: a calcium ion signal channel, a Wnt signal channel, a Hippo signal channel, a TNF signal channel, a VEGF signal channel, a p53 signal channel, an NF-kappa B signal channel, an Ras signal channel, an mTOR signal channel, an RNA degradation signal channel, a PPAR signal channel, a cAMP signal channel, an HIF-1 signal channel, an Insulin signal channel, an AMPK signal channel, an Rap1 signal channel, an apoptosis signal channel, a cell cycle signal channel, a cGMP-PKG signal channel, a DNA copying signal channel, an MAPK signal channel, a PI3K-Akt signal channel and an autophagy signal channel. The polyether compound provided by the invention can cause the tumor cell death through regulating and controlling various signal channels; and a better anti-cancer effect isachieved.

Description

technical field [0001] The invention relates to the field of biomedicine. Specifically, the present invention relates to the use of polyether compounds in the preparation of medicines and the method for screening anticancer medicines. Background technique [0002] Polyether antibiotics are an important class of type I polyketides produced by Streptomyces. Currently, they are mainly used as veterinary drugs to prevent coccidiosis in animals and improve feed utilization to promote growth, such as: salinomycin, lasalocid , nigericin and maduramycin. Up to now, more than 120 kinds of polyether compounds have been found and reported. The polyether compound carrier has chelated metal cations (Na + 、K + , Ca 2+ ), the ability to form a fat-soluble complex, the carrier can diffuse from the outside to the inside of the cell, and return to the outside of the cell or stay on the cell membrane as a metal ion transport carrier to and from the inside and outside of the cell, promoting...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/351A61P35/00G01N33/68
CPCA61K31/351G01N33/6848
Inventor 刘天罡黄敏坚刘然
Owner WUHAN J1 BIOTECH
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