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

A technology of recombining cells and amino acids, which is applied in general/multifunctional contrast agents, medical preparations with non-active ingredients, and medical preparations containing active ingredients, etc. Incomplete cells, etc., to achieve the effect of high specificity, affinity, and high specificity selection

Active Publication Date: 2020-04-17
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the effect of systemic chemotherapy for triple-negative breast cancer is not as good as other types of breast cancer, and systemic chemotherapy drugs usually have strong toxic and side effects and cause great damage to various organs of the human body
At present, the research on triple-negative breast cancer cells is incomplete, and there is a lack of specific targets, which seriously hinders the development of targeted drugs for triple-negative breast cancer cells

Method used

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  • Polypeptide capable of targeting triple-negative breast cancer cells, and application thereof
  • Polypeptide capable of targeting triple-negative breast cancer cells, and application thereof
  • Polypeptide capable of targeting triple-negative breast cancer cells, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Screening Synthesis of Peptides

[0043] Cells were screened using the Negative- / Positive+ method using the phage peptide library. Among them, breast cancer cells of different types, including MDA-MB-453 and MCF-7, are used for negative screening cells to exclude polypeptides that can bind to these negative screening cells; triple negative screening cells are selected for positive screening. Breast cancer cell MDA-MB-231 is used to select polypeptides that can specifically bind to it. There were three rounds of screening. The screening steps are as follows:

[0044]1) MCF-7, MDA-MB-453, SK-BR3, BT-474, T47D were used as negative screening cells, triple-negative breast cancer cell MDA-MB-231 was used as positive screening cells and cultured in RPMI- 1640 medium.

[0045] 2) After negative screen cells were blocked with 0.5% BSA blocking solution for 1 h, the phage random dodecapeptide library (Ph.D.-12 phage display peptide library kit, New England Bio-LABS, USA) was...

Embodiment 2

[0065] Affinity and binding strength experiments

[0066] In this example, flow cytometry and surface plasmon resonance (SPR) were used to analyze the five polypeptides screened in Example 1 (the polypeptides of SEQ ID No.1 to SEQ ID No.5 are represented by polypeptides 1 to 5, respectively) Affinity and binding constant with triple-negative breast cancer cell MDA-MB-231, the specific steps are as follows:

[0067] a. After the five polypeptides were labeled with quantum dots, they were incubated with triple-negative breast cancer cells MDA-MB-231, and the fluorescence intensity was analyzed by flow cytometry to determine the affinity of the five polypeptides with triple-negative breast cancer cells.

[0068] b. Extract MDA-MB-231 membrane protein from triple-negative breast cancer cells.

[0069] c. The MDA-MB-231 membrane protein of triple-negative breast cancer cells was coupled to the SPR chip by using EDC / NHS.

[0070] d. The targeting polypeptides were formulated into ...

Embodiment 3

[0074] Fluorescent probe targeting triple-negative breast cancer cells

[0075] A fluorescent probe specifically targeting triple-negative breast cancer cells, comprising the polypeptide whose amino acid sequence is shown in SEQID No.5 in Example 1 and the fluorescent group FITC coupled to the peptide chain.

[0076] To detect the targeting specificity of the fluorescent probe, the specific steps are as follows:

[0077] 1) Synthesizing a polypeptide linked to a FITC fluorescent group;

[0078] 2) Inoculate triple-negative breast cancer cells MDA-MB-231 and SK-BR3 cells as a control group in a 24-well plate, and wait for the cells to grow to an appropriate density;

[0079] 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;

[0080] 4) Dilute the polypeptide at 1:500, add to...

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Abstract

The invention discloses a polypeptide capable of specifically targeting triple-negative breast cancer cells, and an application thereof. The polypeptide has an amino acid sequence represented by any one of SEQ ID No.1 to SEQ ID No.5. The polypeptide provided by the embodiment of the invention has high specificity and affinity to the triple-negative breast cancer cells, has weak binding capacity toother types of breast cancer cells, shows high-specificity selection to triple-negative breast cancer 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 targeting triple-negative breast cancer 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 1970s to 11.3 / 100,000...

Claims

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Application Information

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IPC IPC(8): C07K7/08A61K47/42A61K47/64A61K45/00A61K49/00A61P35/00A61K31/337A61K47/69
CPCA61K31/337A61K45/00A61K47/42A61K49/0002A61K49/0021A61K49/0056A61K47/64A61K47/6935A61P35/00C07K7/08
Inventor 马岚姜轶珂
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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