A kind of nano therapeutic agent and its preparation method and application
A therapeutic agent, nanotechnology, applied in the field of biomedical nanomaterials, can solve problems such as low therapeutic effect
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Embodiment 1
[0049] Example 1: Synthesis of Sorafenib-glucose oxidase nano-therapeutic agent (SRF@GOx)
[0050]Add 20 mg of glucose oxidase (GOx) to 10 mL of phosphate buffered saline (PBS, pH 7.4) so that the concentration of glucose oxidase in the obtained mixture is 2 mg / mL, stir and add β-mercaptoethanol, and then slowly add 600 μL of 5 mg / mL of sorafenib (SRF, soluble in ethanol), after the dropwise addition, the above mixture was centrifuged at 6000rpm for 5min, and the precipitate was collected for redissolution to obtain sorafenib-glucose oxidase nanoparticles.
[0051] TEM image of the synthesized sorafenib-glucose oxidase nanotherapeutic agent (SRF@GOx) figure 1 shown. from figure 1 It can be seen that the synthesized sorafenib-glucose oxidase nano-therapeutic agent is in the shape of a sphere, and the particle size of the nano-therapeutic agent is between 100nm and 150nm.
Embodiment 2
[0052] Example 2: Analysis of Zeta potential changes before and after synthesis of Sorafenib-glucose oxidase nanotherapeutic agent (SRF@GOx)
[0053] Zeta potential changes of sorafenib, glucose oxidase and sorafenib-glucose oxidase nano-therapeutic agent were detected by Zeta potential meter.
[0054] Zeta potential diagram of the synthesized sorafenib-glucose oxidase nanotherapeutic agent (SRF@GOx) figure 2 shown. from figure 2 It can be seen that Sorafenib is positively charged (about 5.5mV), and glucose oxidase is negatively charged (about-23.2mV), and the potential value of the synthesized Sorafenib-glucose oxidase nano-therapeutic agent is about-8.7mV, It shows that the formed nanoparticles are the product of combination of Sorafenib and glucose oxidase.
Embodiment 3
[0055] Example 3: Identification of Effects of Different Treatments on the Secondary Structure of Glucose Oxidase
[0056] The effect of different treatments on the secondary structure of glucose oxidase was evaluated by circular dichroism spectrometer. The secondary structures of simple glucose oxidase, sorafenib-glucose oxidase nano-therapeutic agent and the enzyme in its release solution were determined respectively. The result is as image 3 Shown, the Sorafenib-glucose oxidase nano-therapeutic agent that forms can not detect the secondary structure of glucose oxidase substantially, but, in the supernatant after Sorafenib-glucose oxidase nano-therapeutic agent releases, can It was detected that the glucose oxidase secondary structure was restored.
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