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Acid-sensitive fusion peptide targeting to tumors and application of acid-sensitive fusion peptide

A fusion peptide and tumor technology, applied in the field of biomedicine, can solve problems such as inability to meet tumor oxygen demand, insufficient supply of nutrients, hypoxia of tumor tissue, etc.

Active Publication Date: 2019-03-15
BEIJING ZEQIN BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The growth of tumors requires a large amount of nutrients, but due to the disorder of tumor neovascularization and its own non-functional capillaries, the supply of nutrients is insufficient and cannot meet the oxygen demand of the tumor's high metabolism, resulting in hypoxia in this part of the tumor tissue.

Method used

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  • Acid-sensitive fusion peptide targeting to tumors and application of acid-sensitive fusion peptide
  • Acid-sensitive fusion peptide targeting to tumors and application of acid-sensitive fusion peptide
  • Acid-sensitive fusion peptide targeting to tumors and application of acid-sensitive fusion peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Synthesis of embodiment 1 fusion peptide

[0084] 1. Prokaryotic expression

[0085] step:

[0086] Strain Construction

[0087] According to the amino acid sequence (SEQ ID NO.20) of the fusion peptide of the present invention, the DNA sequence is designed for full gene synthesis, and the two ends of the synthesis are connected with the enzyme cutting sites Nde I and XhoI sequences, and these two enzymes are used to digest the fusion peptide respectively The encoding nucleic acid and PET28a vector were recovered, the restriction fragments and the vector were recovered, ligated with T4 ligase, transformed into recipient bacteria BL21(DE3), plated and cultured, and clones were picked and sent for sequencing.

[0088] culture induction

[0089] The clones with correct sequencing were cultured (LB medium, 37°C), induced with IPTG when the OD value reached 0.6-0.8, the final concentration of IPTG was 0.5mM, and the bacteria were harvested by centrifugation for 4-6 hours....

Embodiment 2

[0103] Example 2 Localization of Fusion Peptides on Tumor Cells Cultured in Vitro

[0104] 1. Cell culture

[0105] Use DMEM medium containing 10% calf serum and 1.6 million units of gentamycin / ml for A549 cells at 37°C, 5% CO 2 cultured in a cell culture incubator. Cells were passaged at a ratio of 1:1 after confluence.

[0106] 2. Confocal observation positioning

[0107] A549 cells (5x10 5 Cultivate overnight on the coverslip culture dish, discard the culture supernatant, add PBS with pH 6.3 and 7.4 respectively, add the fusion peptide (60 μg / ml) expressed in Example 1, incubate at 37°C for 1 hour, and discard the supernatant for use Wash with PBS of corresponding pH 3 times, add PBS with pH 6.3 and 7.4, add trastuzumab-FITC or TGLA-FITC (control antibody) (both diluted 1:400) and incubate at 37°C for 30 minutes, discard The clear water was washed 3 times with PBS of corresponding pH, and PBS of pH 7.4 was added for confocal observation. Grouping: (1) pH6.3 untreated ...

Embodiment 3

[0113] Example 3 Evaluation of the effect of the fourth domain of Her2-pHLIP and herceptin in treating tumors

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Abstract

The invention discloses an acid-sensitive fusion peptide targeting to tumors and application of the acid-sensitive fusion peptide. The fusion peptide is formed by connecting low-pH inserted peptide with a fourth structural domain of a tumor surface antigen Her2. The fusion peptide can be inserted into a tumor cell membrane in an acidic environment, a fourth structural domain of Her2 is shown on the surface of a cell, and is successfully recognized by trastuzumab as an anti-tumor drug, and the fusion peptide can effectively treat the tumors. By the research achievement of the invention, the shortcoming that an existing anti-tumor drug can only be used for treating one cancer or can only be used for treating one class of one cancer is overcome, the fusion peptide of the invention is labeledon the surface of a tumor, and the treatment effect of the trastuzumab to all cancers is achieved, and thus, the acid-sensitive fusion peptide targeting to the tumors has milestone significance to clinic treatment of the tumors.

Description

technical field [0001] The invention belongs to the field of biomedicine, specifically relates to a fusion peptide, and also relates to the effect of the fusion peptide in targeted therapy of tumors. Background technique [0002] In recent years, with the rapid development of my country's economy, the people's material and cultural level has been continuously improved, and the lifestyle has also undergone tremendous changes. Along with our living environment, changes have also taken place, such as water quality deterioration, air quality decline, and so on. Due to the change of lifestyle and the decline of environmental quality, great changes have taken place in the causes of death in my country. Non-communicable diseases such as malignant tumors, cardiovascular diseases, and chronic diseases have become the main causes of death in my country. The death rate of the country occupies a large proportion and has become a problem that we cannot ignore. [0003] At present, the co...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K47/64A61P35/00
CPCA61K47/64A61P35/00C07K14/195C07K14/4748C07K2319/10
Inventor 王颖魏化伟杨承刚
Owner BEIJING ZEQIN BIOMEDICAL CO LTD
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