Polypeptide capable of specifically targeting triple-negative breast cancer stem cells and application thereof

A technology for recombining cells and amino acids, applied to medical preparations with non-active ingredients, medical preparations containing active ingredients, peptides, etc., can solve incomplete, lack of specific targets, and hinder triple-negative breast cancer stem cells. For drug development and other issues, achieve high specificity and affinity, and high specificity selection effects

Active Publication Date: 2020-04-03
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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  • Polypeptide capable of specifically targeting triple-negative breast cancer stem cells and application thereof
  • Polypeptide capable of specifically targeting triple-negative breast cancer stem cells and application thereof
  • Polypeptide capable of specifically targeting triple-negative breast cancer stem cells and application thereof

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 verifies the expression levels of biomarkers such as SOX2, OCT4, and NANOG in triple-negative breast cancer stem cells to verify whether they have stem cell characteristics.

[0047] figure 1 It is the result of the relative expression of stem cell-related genes SOX2, OCT4, and NANOG in triple-negative breast cancer stem cells (...

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 to SEQ ID No.3 are represented by polypeptides 1 to 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 ...

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) Synthesizing a polypeptide linked to a FITC fluorescent group;

[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 polypeptide at 1:50...

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Abstract

The invention discloses a polypeptide capable of specifically targeting triple-negative breast cancer stem cells and application thereof. The polypeptide has an amino acid sequence shown in any one ofSEQ ID No.1 to SEQ ID No. 3. The polypeptide provided by an embodiment of the invention has high specificity and affinity to the triple-negative breast cancer stem cells, has weak binding capacity toother types of breast cancer cells, shows high specificity and selectivity to the triple-negative breast cancer stem cells, and 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] Breast cancer is the second most common cancer in the world and the most common type of cancer among women. The incidence of breast cancer in my country has increased significantly since the 1990s, and the age of onset is lower than that of European and American countries, and the incidence rate in urban areas is higher than that in rural areas. According to data from my country's tumor registry, the incidence of female breast cancer in my country increased from 37.1 / 100,000 in 2003 to 46.6 / 100,000 in 2013. By 2021, this number is expected to exceed 100 per 100,000 people, and the total number of breast cancer patients nationwide is expected to reach 2.5 million. At the same time, the mortality rate of breast cancer rose from 2.95 / 100,000 in the 197...

Claims

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Application Information

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Patent Type & Authority Applications(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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