Polypeptide or its derivatives and application thereof in preparation of tumor drugs

A technology of derivatives and drugs, which can be used in anti-tumor drugs, drug combinations, and polypeptides containing localization/targeting motifs, etc., can solve problems such as low biological stability, achieve less toxic side effects, safe use, and inhibit EGFR signaling. The effect of pathway activity

Active Publication Date: 2018-09-25
BEIJING WEIFENG YIMIN TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This defect of low biological stability is directly related to the inability of the polypeptide EJ4 to stabilize the α-helical conformation required for its activity in solution

Method used

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  • Polypeptide or its derivatives and application thereof in preparation of tumor drugs
  • Polypeptide or its derivatives and application thereof in preparation of tumor drugs
  • Polypeptide or its derivatives and application thereof in preparation of tumor drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The synthesis of embodiment 1 polypeptide

[0033] For the amino acid sequence of the polypeptide EJ4, see SEQ ID No.1 in the sequence listing. Polypeptide EJ4 was synthesized and purified by Beijing Saibaisheng Gene Technology Co., Ltd.

[0034] Introducing two unnatural amino acids for solid-phase polypeptide chain synthesis. After the solid-phase polypeptide chain is synthesized, ruthenium is used as a catalyst to perform olefin metathesis reaction (RCM) cyclization to obtain the target polypeptide. Finally, the target polypeptide is cleaved from the resin for purification. The steps of solid-phase polypeptide chain synthesis and purification were completed by China Peptide Biochemical Co., Ltd. Wherein, two S-pentenylalanines (S5) are inserted in the i-th and i+4 positions in the amino acid sequence of the polypeptide EJ4, and R-pentenylalanine (R5) and S5 are respectively inserted in the i-th position of the polypeptide EJ4 , i+3 positions; R-octyl alanine (R8)...

Embodiment 2

[0055] Embodiment 2 Circular dichroism method detects the α-helix rate of polypeptide

[0056] The α-helix rate of the polypeptide was detected with a circular dichroism spectrometer (purchased from Jasco, Japan). The polypeptides EJ4, EJ4-S1, EJ4-S2, EJ4-S3, EJ4-S4, EJ4-S5, EJ4-S6, EJ4-S7, EJ4-S8, EJ4-S9, EJ4-S10, EJ4-S11, EJ4-S12, EJ4-S13, EJ4-S14, EJ4-S15, EJ4-S16 and TAT-EJ4 were dissolved in the aqueous solution, and the concentration of the circular dichroism spectrometer was adjusted to 1mg / mL, the results are shown in the table 1. Wherein, the α-helix rate refers to the percentage of the number of peptides of the polypeptide that maintains the secondary structure α-helix to the number of peptides of the total polypeptide.

[0057] Table 1 illustrates that polypeptides EJ4-S1, EJ4-S2, EJ4-S3, EJ4-S4, EJ4-S5, EJ4-S6, EJ4-S7, EJ4-S8, EJ4-S9, EJ4-S10, EJ4-S11, EJ4 -S12, EJ4-S13, EJ4-S14, EJ4-S15, EJ4-S16, and TAT-EJ4 have significantly higher α-helix rates than polypept...

Embodiment 3

[0061] Example 3 Flow Cytometry Detection of Polypeptide Membrane Penetrating Ability

[0062] Flow cytometry measures the ability of polypeptides to cross cell membranes. The specific operation steps are as follows:

[0063] 1. Collect lung cancer cells A549 in the logarithmic growth phase (purchased from the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences), adjust the cell concentration with 1640 medium (purchased from Invitrogen, USA) to make a cell suspension of 200,000 / mL.

[0064] 2. Add 1 mL of the cell suspension prepared in step 1 to a 6-well plate for culture, replace with a new medium after 12 hours, and add 1 μg / mL of the FAM fluorescent group-labeled polypeptide prepared in Example 1 EJ4, EJ4-S1, EJ4-S2, EJ4-S3, EJ4-S4, EJ4-S5, EJ4-S6, EJ4-S7, EJ4-S8, EJ4-S9, EJ4-S10, EJ4-S11, EJ4-S12, EJ4-S13, EJ4-S14, EJ4-S15, EJ4-S16 and TAT-EJ4.

[0065] After 3.6 hours, trypsinization was used to prepare a single cell suspension, and the cells were...

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PUM

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Abstract

The invention discloses a polypeptide to specifically promote degradation of EGFR (epidermal growth factor receptor) protein, or its derivatives, and application of the polypeptide in the preparationof tumor drugs. An amino acid sequence of the polypeptide is shown as in sequence table SEQ ID No. 1; alternatively, two or more amino acids in the amino acid sequence shown as in the sequence table SEQ ID No. 1 are replaced with side-chain-connectable non-natural amino acids; the derivatives include chimeric peptides that are formed by connecting the polypeptide with cell penetrating peptides. The polypeptide or its derivatives can promote degradation of EGFR protein and inhibit EGFR signal pathway activity; therefore, the polypeptide or its derivatives are applicable to the preparation of tumor drugs. Drugs prepared herein are suitable for treating the tumors, such as lung cancer, intestinal cancer, pancreatic cancer, breast cancer, liver cancer, and glioma.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a polypeptide or its derivative and its application in the preparation of drugs for treating tumors. Background technique [0002] Epidermal growth factor receptor (EGFR) is a member of the ErbB transmembrane receptor tyrosine kinase family, also named ErbB1 or HER-1. EGFR is highly expressed or signally hyperactivated in a variety of human tumors. Activated EGFR activates MAPK / ERK, PI3K / Akt and other signaling pathways, and plays a promoting role in tumor proliferation, angiogenesis, tumor metastasis, tumor immune escape, tumor drug resistance, and tumor metabolic reprogramming. In addition to participating in tumor regulation as a membrane receptor activation signal pathway, EGFR can also translocate to the nucleus as a new type of transcription factor, acting independently or in cooperation with other transcription factors in the nucleus on target genes closely relate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K19/00A61K38/10A61P35/00
CPCA61P35/00C07K7/08A61K38/00C07K2319/10A61K38/10A61K45/06A61K47/64C07K19/00
Inventor 胡卓伟花芳
Owner BEIJING WEIFENG YIMIN TECH
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