A polypeptide specifically targeting triple-negative breast cancer stem cells and its application

A triple-negative breast cancer, application technology, applied in the direction of non-active ingredient medical preparations, active ingredient-containing medical preparations, peptides, etc., can solve the problem of incomplete and hindered triple-negative breast cancer stem cell targeted drug research and development, Lack of specific targets and other issues

Active Publication Date: 2021-09-10
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current research on triple-negative breast cancer stem cells is incomplete and lacks specific targets, which seriously hinders the development of targeted drugs for triple-negative breast cancer stem cells

Method used

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  • A polypeptide specifically targeting triple-negative breast cancer stem cells and its application
  • A polypeptide specifically targeting triple-negative breast cancer stem cells and its application
  • A polypeptide specifically targeting triple-negative breast cancer stem cells and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Culture and identification of triple-negative breast cancer stem cells

[0043] A large number of triple-negative breast cancer cells with stem cell characteristics were cultivated by a serum-free low-adsorption suspension culture method, and their stem cell characteristics were verified for subsequent screening of targeted polypeptides.

[0044] The specific operation steps are as follows:

[0045] (1) Use serum-free low-adsorption suspension to culture triple-negative breast cancer stem cells. The medium components used include DMEM / F12 medium, epidermal growth factor, basic fibroblast growth factor, insulin, and B27.

[0046] (2) qRT-PCR verification in triple-negative breast cancer stem cells SOX2 , OCT4 , NANOG Wait for the expression level of the biomarker to verify whether it has stem cell characteristics.

[0047] figure 1 It is a stem cell-related gene in triple-negative breast cancer stem cells (CSC) and triple-negative breast cancer cell line MDA-MB...

Embodiment 2

[0071] In this example, flow cytometry and surface plasmon resonance (SPR) were used to analyze the three polypeptides screened in Example 1 (the polypeptides of SEQ ID No.1~SEQ ID No.3 are represented by polypeptides 1~3 respectively) Affinity and binding constants with triple-negative breast cancer stem cells, the specific steps are as follows:

[0072] a. After the three polypeptides were labeled with quantum dots, they were incubated with triple-negative breast cancer stem cells, and the fluorescence intensity was analyzed by flow cytometry to determine the affinity of the three polypeptides with triple-negative breast cancer stem cells.

[0073] b. Extract the membrane protein of triple-negative breast cancer stem cells.

[0074] c. Using EDC / NHS to couple triple-negative breast cancer stem cell membrane proteins to the SPR chip.

[0075] d. The targeting polypeptides were formulated into different concentrations, and flowed through the surface of the SPR chip in sequenc...

Embodiment 3

[0079] Fluorescent probe targeting triple-negative breast cancer stem cells

[0080] A fluorescent probe specifically targeting triple-negative breast cancer stem cells, comprising the polypeptide whose amino acid sequence is shown in SEQ ID No.1 in Example 1 and the fluorescent group FITC coupled to the peptide chain.

[0081] To detect the targeting specificity of the fluorescent probe, the specific steps are as follows:

[0082] 1) Synthesis of peptides linked to FITC fluorescent groups;

[0083] 2) Inoculate triple-negative breast cancer stem cells and MDA-MB-231 cells as a control group in a 24-well plate, and wait for the cells to grow to an appropriate density;

[0084] 3) Aspirate the medium, wash with PBS three times, each time for 3 minutes, add 500 μL of 4% paraformaldehyde solution to each well, fix at room temperature for 20 minutes, absorb the fixative, wash with PBS solution three times, each time for 3 minutes;

[0085] 4) Dilute the peptide at 1:500, add it ...

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PUM

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Abstract

The invention discloses a polypeptide capable of specifically targeting triple-negative breast cancer stem cells and the application of the polypeptide. The polypeptide has the amino acid sequence shown in any one of SEQ ID No.1 to SEQ ID No.3. The polypeptide provided by the embodiment of the present invention has high specificity and affinity with triple-negative breast cancer stem cells, but has relatively weak binding ability with other types of breast cancer cells, showing high specificity for triple-negative breast cancer stem cells, It can be used for targeted therapy or imaging diagnosis of triple-negative breast cancer.

Description

technical field [0001] The invention relates to the field of medicinal chemistry, in particular to a polypeptide specifically targeting triple-negative breast cancer stem cells and its application. Background technique [0002] According to different molecular types, breast cancer can be divided into: estrogen / progesterone positive type (ER / PR+), HER2 high expression type (HER2+) and triple negative type. Triple-negative breast cancer is a highly aggressive and highly heterogeneous subtype of breast cancer that is negative for estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2 by immunohistochemistry. Although the incidence of triple-negative breast cancer accounts for only 12-20% of the total incidence of female breast cancer, its age of onset is young, its aggressiveness is high, its prognosis is extremely poor, and its survival rate is low. The 5-year survival rate of advanced patients is less than 15%. For ER / PR+ breast cancer, endoc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C12N15/11A61K47/64A61K31/337A61P35/00
CPCA61K31/337A61K47/64A61P35/00C07K7/08
Inventor 马岚姜轶珂
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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