Polypeptide aggregate used for regulating subtype transformation of macrophages and preparation method and application thereof

A macrophage and aggregate technology, which is applied in the field of regulating the transformation of M2 macrophages into M1 macrophages and the preparation of polypeptide aggregates, can solve the problems of low macrophage efficiency and the like

Active Publication Date: 2020-10-23
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the methods that can achieve macrophage subtype

Method used

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  • Polypeptide aggregate used for regulating subtype transformation of macrophages and preparation method and application thereof
  • Polypeptide aggregate used for regulating subtype transformation of macrophages and preparation method and application thereof
  • Polypeptide aggregate used for regulating subtype transformation of macrophages and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] This embodiment provides a polypeptide aggregate regulating macrophage subtype transformation and a preparation method thereof.

[0071] The polypeptide molecule is composed of three functional domains, including an aggregation driving unit, a shape transformation driving unit and a targeting unit.

[0072] 1. Design of polypeptide molecules

[0073] In order to realize the site-specific reassembly of polypeptide aggregates on specific cell membranes, the targeted polypeptide fragments and the morphology conversion drive unit that can be targeted to M2 macrophages are integrated through the amide bond to realize the integration of the two functional segments, and secondly, the bipyrene The condensation reaction between the carboxyl group of the (bispyrene, BP) molecule and the amino group of the side chain couples the hydrophobic small organic molecule bispyrene to the polypeptide backbone to obtain the final target molecule.

[0074] The final target polypeptide aggre...

Embodiment 2

[0087] This example studies the assembly behavior of the polypeptide molecules prepared in Example 1 in solution.

[0088] The principle of the experiment is: taking the difference between the ultraviolet absorption spectrum and the fluorescence emission spectrum of bispyrene molecules in the dispersed and aggregated states as a theoretical basis, to obtain their spectral properties in different solvents, and to explore their assembly behavior.

[0089] The specific steps are as follows: water is used as a poor solvent and dimethyl sulfoxide (DMSO) is used as a good solvent, and they are mixed according to different volume ratios to provide assembly environments with different polarities.

[0090] Configure the molar concentration of BP-pep1 to be 3×10 -5 M solution, the ultraviolet absorption spectra of polypeptide molecules in different polar environments were collected by ultraviolet absorption spectrometer.

[0091] like image 3 As shown in the ultraviolet absorption sp...

Embodiment 3

[0098] This example evaluates the efficiency of macrophage subtype transformation induced by polypeptide aggregates at the cellular level.

[0099] M2 macrophages RAW264.7 were incubated with 20 ng / μL interleukin-12 (IL-12) and 30 μM BP-pep1 polypeptide molecules for 2 hours, and the surface of M1 macrophages was identified by flow cytometry. landmark.

[0100] The results of cell flow cytometry were as follows: Figure 6(a) ~ Figure 6(c) As shown, compared with the control group (as shown in Figure 6(a)), IL-12 (as shown in Figure 6(b)) and the polypeptide aggregate BP-pep1 (as shown in Figure 6(c)) were The M1 macrophage markers on the surface of the incubated RAW264.7 cells were 1.90% and 2.51%, respectively, while the expression level of the control group was only 0.67%. The amount is higher than that of the control group, indicating that both can transform macrophages into M1 type.

[0101] At the same time, compared with RAW264.7 co-incubated with IL-12, the expressio...

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Abstract

The invention provides a polypeptide aggregate used for regulating subtype transformation of macrophages and a preparation method and application thereof. The polypeptide aggregate comprises a targeting unit, a morphology transformation driving unit and an aggregation driving unit which are connected in sequence by amide bonds; the targeting unit includes a polypeptide sequence targeting a type M2macrophage. The polypeptide aggregate adopts a segmented design, and the targeting unit targeting the type M2 macrophage, the morphology transformation driving unit and the aggregation driving unit are integrated into a polypeptide molecule, then efficient assembly of the polypeptide aggregate in vitro and precise targeting and site-selective reassembly of the polypeptide aggregate in vivo are realized, transformation of the type M2 macrophage to a type M1 macrophage is realized, and meanwhile, the specificity, precision and efficiency of subtype transformation are guaranteed. Meanwhile, thepolypeptide aggregate has good biocompatibility, and is regarded as a molecule for realizing subtype transformation of the macrophages, and has higher biological safety.

Description

technical field [0001] The present invention relates to the field of medical technology, in particular to a polypeptide aggregate for regulating the transformation of macrophage subtypes and its preparation method and application, especially to a polypeptide for regulating the transformation of M2 macrophages into M1 macrophages Aggregates and methods for their preparation and use. Background technique [0002] During the occurrence and development of tumors, tumor-associated macrophages (TAMs) play a leading role in regulating tumor-associated inflammation. Tumor-associated macrophages can adopt different subtypes in a dynamic and reversible manner according to their microenvironment, regulate the body's innate and adaptive immune responses, and ultimately maintain the immune balance in the body. The polarization typing of macrophages is the requirement of the diversity of the body's immune function, and its polarization typing is related to the microenvironment and diseas...

Claims

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Application Information

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IPC IPC(8): C07K14/00C07K1/04C07K1/06C12N5/0786
CPCC07K14/00C12N5/0645C12N2501/998Y02P20/55
Inventor 王浩王羿穆合塔尔江·马木提
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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