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Tumor cell nano-gold vesicles, immune nano-gold vesicles, preparation method and application

A tumor cell and nano-gold technology, applied in the field of anti-tumor drugs and carriers, can solve the problems of ineffective inhibition of tumor metastasis and recurrence, potential safety hazards, and weak killing effect

Active Publication Date: 2020-10-30
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention solves the technical problems of potential safety hazards in the traditional preparation process of gold nanoparticles in the prior art, weak killing effect in tumor treatment, and inability to effectively inhibit tumor metastasis and recurrence

Method used

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  • Tumor cell nano-gold vesicles, immune nano-gold vesicles, preparation method and application
  • Tumor cell nano-gold vesicles, immune nano-gold vesicles, preparation method and application
  • Tumor cell nano-gold vesicles, immune nano-gold vesicles, preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Preparation of tumor cell nano-gold vesicles

[0039]The specific synthesis steps are as follows: add chloroauric acid solution (0.5mM) to the B16F10 cells grown to 80% and starve and mix evenly, place at 37 ° C, 5% CO 2 Cultivate in a culture environment for 4 days, then suck out the supernatant, centrifuge at 600g for 10 minutes to remove cell debris, repeat twice, collect the supernatant at 10,000g, centrifuge for 30 minutes to collect and purify the gold nanovesicles, and centrifuge the supernatant twice The second time, the collected pellets were combined and resuspended, washed with 1 ml of phosphate buffer at 600 g and centrifuged at a low speed for 10 minutes, and the supernatant was the tumor cell gold nanovesicles.

[0040] The shape of nanoparticles was observed by optical microscope and transmission electron microscope. It can be seen that the gold nanovesicles exocytized by tumor cells are purple-red. figure 1 The gold nanovesicles showing exocy...

Embodiment 2

[0043] Embodiment 2: Preparation of nano gold vesicles with immune function

[0044] The specific operation steps for the preparation of nano-gold vesicles are as follows: add 20 micrograms of tumor cell gold nano-vesicles prepared in Example 1 to the blank medium of dendritic cells DC2.4 grown to 80%, incubate for 6 hours and then replace For a new blank medium, irradiate under 8W UV light for 1 hour, then incubate at 37°C, 5% CO 2 Continue to culture in the environment. After 24 hours, collect the supernatant, 600g, 10 minutes of low-speed centrifugation to remove cell debris, repeat twice, and then 13000g of supernatant, 30 minutes of high-speed centrifugation to collect purified gold nanovesicles, supernatant repeated centrifugation Twice, the combined precipitates were resuspended in 250 microliters of phosphate buffer, washed by centrifugation at 600g for 10 minutes to obtain gold nanovesicles with immune function after being treated by immune cells.

[0045] Through th...

Embodiment 3

[0049] Example 3: Study on the in vitro photothermal conversion effect of nano-gold vesicles

[0050] In order to investigate the light-to-heat conversion ability of immune gold nanovesicles, we conducted a light-heating experiment on gold nanovesicles with an Au concentration of 100ug / mL. The specific steps are: set the power to 2W / cm 2 The near-infrared light (NIR) directly irradiates the nano-gold vesicle solution, and an infrared camera is used to monitor the temperature change of the solution within 3 minutes. The results are shown in Figure 8, which shows that the nano-gold vesicles have good light-to-heat conversion ability. Figure 8(A) is the infrared photo of the photothermal conversion of the immune nano-gold vesicles; Figure 8(B) is the corresponding temperature rise curve of the immune nano-gold vesicles. The thermal killing temperature can realize the killing effect of light and heat on tumor cells.

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Abstract

The invention discloses a tumor cell nanogold vesicle and a preparation method and application thereof, and a tumor cell immune nanogold vesicle and a preparation method and application thereof. The tumor cell nanogold vesicle adopts a core-shell structure consisting of an outer-layer vesicle and an inner core, the outer-layer vesicle is a cell membrane wrapping the inner core, the inner core consists of nanogold particles, and the outer-layer vesicle carries a tumor cell specific antigen. The tumor cell immune nanogold vesicle carries a peptide segment having the tumor cell specific antigen, and reserves characteristics of immunocyte. According to the nanogold vesicle generated through tumor cells, provided by the invention, on the base of keeping original thermal treatment of nanogold, the tumor specific antigen carrying mother cells is given to the vesicle, an organism is stimulated to produce immune response, relevant immune cells and factors are released for performing immunization killing on tumors, besides, system immune defence is activated, tumor metastasis is restrained, and tumor recurrence is restrained in a long-acting manner. The carried antigen information can be further presented to the immunocyte after being treated through antigen presenting cells, so that immune response is induced, and besides, immune treatment and thermal treatment of the tumor are realized.

Description

technical field [0001] The present invention relates to the field of anti-tumor drugs and carriers, more specifically, to a nano-gold vesicle exocytosed by tumor cells that carries tumor antigens and has light-to-heat conversion capabilities, immune nano-gold vesicles, preparation methods and applications thereof . Background technique [0002] Gold nanoparticles have high photothermal conversion ability, good stability, low toxicity and adjustable surface properties, and are widely used in various treatments of tumors, such as tumor photothermal therapy, tumor photodynamic therapy, tumor diagnosis and anti-tumor therapy. Delivery of tumor drugs, etc. However, a variety of chemical reagents including sodium borohydride and other toxic reagents need to be added in its chemical preparation process, which greatly limits its safe production and clinical application. Moreover, the drugs based on gold nanoparticles studied in recent years are mostly used in single-method anti-tu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/00A61K41/00A61K9/127A61P35/00
CPCA61K9/1273A61K39/0011A61K41/0052A61P35/00A61K2300/00
Inventor 张志平章丹
Owner HUAZHONG UNIV OF SCI & TECH
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