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Macromolecular temperature-sensitive driver and preparation method and application thereof

A technology of macromolecules and actuators, which is applied in the field of macromolecular temperature-sensitive actuators and its preparation, and can solve problems such as application limitations, complicated processes, and inability to realize receptor protein regulation.

Active Publication Date: 2017-08-08
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Biologically active ligands cannot manipulate receptor proteins in time and space
Optogenetics relies on transgenic technology to transfect light-sensitive proteins in cells. The process is complicated and the application is limited. It cannot realize the regulation of receptor proteins with poor motility
Although the magnetic regulation method can realize the active regulation of the receptor protein in time and space, this method relies on complex instrument processing design
[0003] CN106397577A discloses a dual-stimuli-responsive collagen polypeptide polymer. The polymer has a collagen polypeptide as the main chain, and the amino groups on its side chains are grafted with alkoxy ether dendrons through dynamic linkage reactions. The prepared collagen polypeptide polymer has dual response to temperature and pH, but the material is not based on the temperature and pH response of the cell membrane surface, and cannot be directly used for regulation at the cell level

Method used

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  • Macromolecular temperature-sensitive driver and preparation method and application thereof
  • Macromolecular temperature-sensitive driver and preparation method and application thereof
  • Macromolecular temperature-sensitive driver and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The structural schematic diagram of the macromolecular temperature-sensitive actuator prepared in this example is as follows: figure 1 As shown, it includes a targeting module composed of a polypeptide ligand, a driving module composed of a temperature-sensitive polymer, and a trigger module composed of a photothermal conversion molecule, the trigger module is connected to the targeting module, and the targeting module is The module is connected with the drive module. Among them, the photothermal conversion molecule is rhodopsin-18, and the polypeptide ligand is CKGGMSRTMS (in order to connect rhodopsin-18, amino acid K is added; in order to connect the polypeptide with the polymer, amino acid C is added. GG is a flexible spacer), connected with The molecular structure of the polypeptide ligand CK(P18)GGMSRTMS of rhodopsin-18 is shown below:

[0050]

[0051] In this embodiment, the synthesis method of the macromolecular temperature-sensitive actuator is as follows:...

Embodiment 2

[0067] The polypeptide ligand CK(P18)GYHWYGYTPQNVI was synthesized using the same polypeptide ligand synthesis method as in step (1) of Example 1, and its structure is as follows:

[0068]

[0069] Use the same method as step (2) of Example 1 to connect rhodopsin-18 to the polypeptide ligand, and then perform the same steps (3) and (4) as Example 1 to prepare a macromolecular thermosensitive actuator , whose structure is as follows:

[0070]

[0071] The MALDI-TOF spectrum of the polypeptide ligand CK(P18)GYHWYGYTPQNVI prepared in this example is as follows Figure 4 As can be seen from the figure, the molecular weight of the polypeptide is 2375.0, and the peak of the molecule plus sodium ion is 2397.0.

[0072] The structure of the obtained macromolecular thermosensitive actuator was characterized by hydrogen NMR spectroscopy, and the results are as follows Figure 5 As can be seen from the figure, the chemical shift values ​​δ=3.43ppm and δ=3.18ppm correspond to -OCH...

Embodiment 3

[0074] The macromolecular temperature-sensitive actuator prepared in Example 1 is Image 6 The shown method regulates the oligomerization of the membrane receptor protein on the cell membrane surface, and the driver pulls the oligomerization of the receptor protein to verify that the method is as follows:

[0075] FITC-labeled thermosensitive driver molecules were incubated with adherent SKBR3 cells at a concentration of 3 μM in a confocal dish for half an hour to allow the molecules to bind to cell surface receptor proteins; the results of confocal laser microscopy imaging are as follows Figure 7As shown, picture A is the SEM picture without laser irradiation, and picture B is the SEM picture with and without laser irradiation. When the laser is not irradiated, the driver molecules are in a stretched state and spread on the cell surface, and almost There is no cell entry, and the cell surface fluorescence is weak; when the wavelength of 655nm is used, 4.77W cm -2 The sample...

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Abstract

The invention provides a macromolecular temperature-sensitive driver and a preparation method and application thereof. The macromolecular temperature-sensitive driver comprises targeting modules composed of polypeptide ligands, a driving module composed of a temperature-sensitive polymer, and triggering modules composed of photothermal conversion molecules, the triggering modules are connected to the targeting modules, and the targeting modules are connected to the driving module. The driver can be targeted to cell membrane protein by means of the polypeptide ligands, and can be effectively specifically combined with the membrane protein by way of active targeting of molecular recognition, the surface of the cell membrane is partially heated to activate the driver, so that the driver is collapsed and aggregated to generate pulling force, consequently, the assembly control of different membrane receptor proteins is realized, and a new effective strategy is provided for the research on signaling pathway, immunocyte activation, tumor treatment and the like.

Description

technical field [0001] The invention belongs to the technical field of polymer materials, and relates to a macromolecular temperature-sensitive actuator and a preparation method and application thereof. Background technique [0002] Cell membrane receptor proteins can recognize and bind specific biologically active ligands to form complexes, thereby activating and initiating a series of physicochemical changes, which in turn lead to the final biological effects of the substances. These biological effects include cell proliferation, cell differentiation, apoptosis, and the like. There are three main methods for regulating the oligomerization of membrane receptor proteins: 1) using biologically active ligands; 2) optogenetics; 3) magnetic regulation. But each method has its own drawbacks. Biologically active ligands cannot achieve spatial and temporal manipulation of receptor proteins. Optogenetics relies on transgenic technology to transfect light-sensitive proteins in cel...

Claims

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Application Information

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IPC IPC(8): C08F220/54C08F220/58C08F8/34C08F8/30A61K47/42
CPCA61K47/42C08F8/34C08F8/30C08F220/54
Inventor 王浩乔圣林马洋
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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