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Application of a composition of fullerene C60 nanocrystals combined with camkii inhibitors

A technology of nanocrystals and fullerenes, applied in the field of biomedicine, can solve problems that have not been reported before, and achieve good effects, high inhibition efficiency, and good application prospects

Active Publication Date: 2021-07-27
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports about the application of fullerene C60 nanocrystals on osteosarcoma cells and its effect on osteosarcoma cells CaMKIIα, and whether it is possible to promote the anti-osteosarcoma effect of C60 nanocrystals by regulating the activity of CaMKII never reported

Method used

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  • Application of a composition of fullerene C60 nanocrystals combined with camkii inhibitors
  • Application of a composition of fullerene C60 nanocrystals combined with camkii inhibitors
  • Application of a composition of fullerene C60 nanocrystals combined with camkii inhibitors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Composition of fullerene C60 nanocrystals combined with CaMKII inhibitors (1)

[0039] 1. Experimental materials

[0040] Fullerene C60 powder, KN-93 (Cat.No.K1385) was purchased from Sigma Company.

[0041] 2. Experimental method

[0042] Water-soluble C60 nanoparticles (nano-C60) were synthesized by tetrahydrofuran-to-water method, and its preparation method was as follows:

[0043] (1) Dissolve 5 mg of nano-C60 powder in 500 mL of unopened tetrahydrofuran, and stir at room temperature for 24 hours in the dark;

[0044] (2) Use a 0.45 μm filter membrane to filter out undissolved C60 particles, stir the filtrate on a magnetic stirrer, and add an equal volume of deionized water at a speed of 1 L / min to transfer it into the water phase;

[0045] (3) The tetrahydrofuran in the solution was removed by rotary evaporation, and the solution was concentrated to 50 mL by further evaporation to synthesize fullerene C60 nanocrystals.

[0046] 3. Experimental results...

Embodiment 2

[0048] Example 2 Effect of C60 nanocrystals on the viability of osteosarcoma cells (2)

[0049] 1. Experimental materials

[0050] FullereneC60 powder, KN-93 (Cat.No.K1385) was purchased from Sigma; Calmodulin (Calmodulin, CaM, 208690) was purchased from Calbiochem, USA; CaMKII activity detection kit (CY-1173) was purchased from Japan CycLex Company; pcDNA3-AKT-T7 and pcDNA3-CA-AKT-T7 plasmids were purchased from Changsha Youbao Biological Company; CaMKII antibody (Cat.No.ab52476) and phospho-T286CaMKII antibody (Cat.No.ab32678) were purchased from Abcam Company; Cleaved-PARP, cleaved-caspase3, Blc-2 antibodies were purchased from Cell Signaling Technology Company; GAPDH antibody (Cat.No.MAB374) was purchased from Millipore Company; cell-related culture reagents were purchased from Invitrogen Company.

[0051] 2. Experimental method

[0052] (1) Preparation and characterization of fullerene C60 nanocrystals

[0053] Tetrahydrofuran (THF) to water method, that is, first diss...

Embodiment 3

[0060] Example 3 C60 nanocrystals activate osteosarcoma cells CaMKIIα (3)

[0061] 1. Experimental materials

[0062] FullereneC60 powder, KN-93 (Cat.No.K1385) was purchased from Sigma; Calmodulin (Calmodulin, CaM, 208690) was purchased from Calbiochem, USA; CaMKII activity detection kit (CY-1173) was purchased from Japan CycLex Company; pcDNA3-AKT-T7 and pcDNA3-CA-AKT-T7 plasmids were purchased from Changsha Youbao Biological Company; CaMKII antibody (Cat.No.ab52476) and phospho-T286CaMKII antibody (Cat.No.ab32678) were purchased from Abcam Company; Cleaved-PARP, cleaved-caspase3, Blc-2 antibodies were purchased from Cell Signaling Technology Company; GAPDH antibody (Cat.No.MAB374) was purchased from Millipore Company; cell-related culture reagents were purchased from Invitrogen Company.

[0063] 2. Experimental method

[0064] (1) Preparation and characterization of fullerene C60 nanocrystals

[0065] Tetrahydrofuran (THF) to water method, that is, first dissolve 5 mg of ...

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Abstract

The invention relates to the application of a fullerene C60 nanocrystal combined with a CaMKII inhibitor in osteosarcoma. The application is to enhance the ability of the C60 nanocrystal to kill osteosarcoma cells and reduce the Dosage of crystals. The experimental results showed that: (1) Inhibiting the activity of CaMKIIα could enhance the ability of C60 nanocrystals to kill osteosarcoma cells. (2) The dosage of C60 nanocrystals can be reduced by combining C60 nanocrystals with CaMKII inhibitors. (3) The combination of C60 nanocrystals and CaMKII inhibitor enhances the killing of osteosarcoma cells by inhibiting the AKT signaling pathway.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of a combination of fullerene C60 nanocrystals combined with CaMKII inhibitors. Background technique [0002] Due to its remarkable geometric structure, fullerene C60 has many unique physical and chemical properties (photo-induced oxygen free radicals, active chemical reaction ability, etc.), and has a very wide range of applications in the field of biomedicine. For example, C60 can generate singlet oxygen, cut DNA, induce autophagy, and regulate immunity. And because of the active π-bond structure on the surface of C60, it is easy to introduce other structures on its surface, resulting in a large number of C60 derivatives. Derivatives of C60 also have outstanding applications in drug delivery, gene therapy, and tumor targeting. [0003] CaMKII is a class of calcium ion / calmodulin (Ca 2+ / CaM)-dependent protein kinases, mainly including several subtypes suc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K33/44A61K9/14A61K45/06A61P35/00
CPCA61K9/14A61K33/44A61K45/06A61P35/00A61K2300/00
Inventor 许婧尹飞王卓莹华莹奇蔡郑东
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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