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Anti-tumor polypeptide and application thereof in preparing anti-tumor drugs

An anti-tumor drug and anti-tumor technology, applied in the field of biomedicine, can solve the problems of poor prognosis of pancreatic cancer patients, and achieve the effects of strong selectivity, wide clinical application value, and low dosage

Active Publication Date: 2019-10-15
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Clinical studies have proved that NRP-1 is highly expressed in pancreatic cancer tissues, and its expression level is positively correlated with the clinical stage, lymph node invasion, distant organ metastasis, and pathological stage of pancreatic cancer; and the prognosis of pancreatic cancer patients with high expression of NRP-1 is poor

Method used

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  • Anti-tumor polypeptide and application thereof in preparing anti-tumor drugs
  • Anti-tumor polypeptide and application thereof in preparing anti-tumor drugs
  • Anti-tumor polypeptide and application thereof in preparing anti-tumor drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The preparation of embodiment 1 TMD6 polypeptide

[0056] Proceed as follows:

[0057] 1. Resin swelling: Weigh 0.6g of CTC resin with a degree of substitution of 1.0mmol / g, put it into a reaction tube, add DCM (15ml / g), and shake for 30min.

[0058] 2. Deprotection: remove DMF, add 20% piperidine DMF solution (15ml / g), shake for 5min, remove and add 20% piperidine DMF solution (15ml / g), shake for 15min.

[0059] 3. Detection: Take out the piperidine solution, take a small amount of resin (about 20 resins), wash it three times with ethanol, add one drop of ninhydrin, KCN, and phenol solution, heat (105-110°C) for 5 minutes, and it turns blue .

[0060] 4. The first wash: Wash twice with DMF (10ml / g), then twice with methanol (10ml / g), and finally twice with DMF (10ml / g).

[0061] 5. Condensation: Three-fold excess of protected amino acids and three-fold excess of HBTU were dissolved with as little DMF as possible, added to the reaction tube, immediately added with te...

Embodiment 2

[0068] Example 2 Cell Viability Detection Experiment

[0069] Human umbilical vein endothelial cells were seeded in 96-well culture plates, 1×10 per well 4 Cells were cultured in ECM (Endothelial cell medium) medium containing 5% fetal bovine serum (FBS), added VEGF (100ng / ml) and TMD1, TMD2, TMD3, TMD4, TMD5, TMD6 , TMD7 or TMD8 eight kinds of polypeptides (the concentration is 50nM). Pancreatic cancer, gastric cancer and cholangiocarcinoma cells were routinely inoculated in 96-well culture plates, 5×10 per well 3 Cells were cultured in DMEM (Dulbecco's modified Eagle's medium) medium containing 10% fetal bovine serum (FBS). at 37°C and 5% CO 2 conditions, incubated overnight. Add 100 microliters of culture solution containing different concentrations of TMD6 polypeptides to each well. After culturing for 24 hours, add 110 microliters of fresh culture medium (10 microliters containing CCK-8 solution) to each well, shake the 96-well plate slightly for 10 seconds, then put...

Embodiment 3

[0072] Example 3 Cell Apoptosis Detection Experiment

[0073] Pancreatic cancer, gastric cancer and cholangiocarcinoma cells were routinely inoculated in 8-well culture plates, 1×10 per well 5 Cells were cultured in DMEM (Dulbecco's modified Eagle's medium) medium containing 10% fetal bovine serum (FBS). at 37°C and 5% CO 2 conditions, incubated overnight. Add 100 microliters of culture solution containing different concentrations of polypeptides to each well. After 24 hours of culture, the cells were collected for detection of apoptosis.

[0074] Image 6 B. Image 6 C. Figure 7 B. Figure 7 C. Figure 8 B and Figure 8Cells in C were detected by Annexin V-FITC apoptosis detection kit. Take 50,000 to 100,000 resuspended cells, centrifuge at 1000×g for 5 minutes, discard the supernatant, and add 195 μL Annexin V-FITC conjugate solution to gently resuspend the cells. Add 5 μL Annexin V-FITC and incubate at room temperature for 10 minutes in the dark. Centrifuge at ...

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Abstract

The present invention discloses an anti-tumor polypeptide and an application thereof in preparing anti-tumor drugs. Aiming at a molecular structure, a ligand binding mode and amino acid sequence characteristics of a NRP-1 transmembrane region, and the transmembrane region capable of specifically binding the human source NRP-1 molecular structure and binding ligands is designed to block binding ofNRP-1 with a variety of the ligands of VEGF, EGF, and HGF, thereby blocking the anti-tumor polypeptide drugs of downstream cellular signaling pathways. The anti-tumor polypeptide binds to the NRP-1 transmembrane region monomer competitively, interferes dimerization of NRP-1, inhibits the binding of the NRP-1 to the ligands, thus inhibits activation of various tumor cell signaling pathways, and achieves purposes of inhibiting tumor cell proliferation, migration and infiltration, and promoting apoptosis. The provided anti-tumor polypeptide provides a new technical means for treatment of tumors.

Description

technical field [0001] The invention relates to an anti-tumor polypeptide and its application in the preparation of anti-tumor drugs, in particular to a polypeptide targeting solid tumor cell membranes with high expression of NRP-1 and its application. The invention belongs to the technical field of biomedicine. Background technique [0002] Molecular targeted drugs have become the focus and development direction of anticancer drug research. They have made gratifying progress in the treatment of various tumors and have been put into clinical use one after another. However, the research and development of targeted drugs for pancreatic cancer, gastric cancer and cholangiocarcinoma among the high-incidence cancer types in my country is relatively lagging behind. According to the latest data released by the National Cancer Center of my country, the incidence of pancreatic cancer has reached the 10th, and the mortality rate has risen to the 6th; the incidence of gastric cancer r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K38/08A61P35/00
CPCA61K38/00A61P35/00C07K7/06
Inventor 孙学英马立新李伟东姜宪乔海泉
Owner HARBIN MEDICAL UNIVERSITY
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