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Nano vaccine as well as preparation method and application thereof

A nano-vaccine and nano-particle technology, applied in the field of biomedicine, can solve problems such as poor solubility and application limitations, and achieve the effects of low toxicity and side effects, large drug loading, and significant lethality.

Active Publication Date: 2022-03-15
UNIV OF ELECTRONIC SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the poor solubility of NLG-919 and BLZ-945, their application in therapeutic delivery is greatly limited

Method used

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  • Nano vaccine as well as preparation method and application thereof
  • Nano vaccine as well as preparation method and application thereof
  • Nano vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Preparation of nano-vaccine, verification of pH-responsiveness of nano-vaccine

[0036] A nano-vaccine in response to acid stimulation, which is prepared through the following steps:

[0037] (1) Pour 5 mL of chloroform into a clean round bottom flask with a graduated cylinder, then add PEG-hyd-PCL (25 mg) and PEI-PCL (12.5 mg) into the round bottom flask, add magnets and stir on a magnetic stirrer until dissolved; after fully dissolved, evaporate in a rotary evaporator at 60°C. After the chloroform was completely evaporated to form a thin film, 3ml of deionized water at a temperature of 65°C was slowly dropped into it while stirring, and further stirred at room temperature for 4h, then dialyzed and washed to obtain unloaded hybrid micelles (HM).

[0038](2) Accurately weigh 0.25mg BLZ-945 and 0.5mg NLG-919 and place them in a centrifuge tube, add 0.05mL DMSO, vortex to fully dissolve, then add 1mg of HM prepared in step (1) to the solution , mixed evenly, ...

Embodiment 2

[0041] Example 2: In vitro anti-tumor activity of nanomicelle vaccine complex (BN@HM-OVA) and its cellular immune response analysis

[0042] (1) Antigen uptake by myeloid-derived dendritic cells (BMDCs)

[0043] Whether DCs cells can successfully uptake antigens is the key to promoting their maturation. Mouse bone marrow cells were induced to BMDCs on day 5 in vitro, and BMDCs were stained with cell membrane dye Dil. According to 1×10 6 / mL BMDCs The stained BMDCs were co-incubated with 10 μL BN@HM-OVA-FITC (OVA-FITC concentration 10 μg / mL) in different pH (pH 7.4, pH 6.8) media for different periods of time (2h, 6h). The uptake of BN@HM-OVA-FITC by BMDCs was detected by laser scanning confocal microscopy.

[0044] From image 3 The respective confocal images showed that BN@HM-OVA-FITC could be efficiently taken up by BMDCs in a time-dependent manner after 2 h and 6 h incubation at pH 7.4 and 6.8. Furthermore, regardless of the incubation time, BMDCs at pH 6.8 internalize...

Embodiment 3

[0051] Embodiment 3: nano vaccine tumor live treatment study

[0052] (1) Anti-tumor effect of nano-vaccine complexes in prophylactic administration mode on tumor-bearing mice

[0053] To evaluate the effectiveness of the nanovaccine in tumor prevention, six-week-old C57BL / 6 mice were immunized using a homologous prime-boost regimen. Mice were divided into six groups (n=6 per group), one group was injected with Saline, one group was injected with Free OVA, and four groups were injected with HM-OVA for immunization (OVA: 5 nmol per mouse), and at designated time points Injected subcutaneously into both footpads of mice. Three weeks after the last immunization, each mouse was subcutaneously inoculated with 2×10 5 E. G7-OVA cells. Starting on the 6th day after tumor inoculation, every 4 days, 3 groups of mice immunized with HM-OVA were injected with Free NLG-919, FreeBLZ-945 and Free NLG-919+Free BLZ-945, total Dosing 5 times. Tumor growth was monitored every two days using ...

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Abstract

The invention discloses a nano vaccine and a preparation method and application thereof.The nano vaccine prepared through the method can sequentially respond to a tumor weak acid microenvironment, can remodel a tumor immunosuppressive microenvironment and can carry various therapeutic agents and antigens, a new thought is provided for design of a nano vaccine platform, and the nano vaccine has the advantages of being high in practicability and good in application prospect. The invention can be used for combined immunotherapy of tumors.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a nano vaccine and its preparation method and application. Background technique [0002] In recent years, cancer immunotherapy has shown great potential to improve the efficacy of cancer treatment. Among different types of cancer immunotherapy strategies, cancer vaccines have been extensively studied for their ability to elicit tumor-specific immune responses. Although cancer vaccines have shown some degree of anti-tumor efficacy in some cases, they have been affected by rapid clearance of soluble antigens, insufficient antigen internalization by antigen-presenting cells (APCs), and inefficient maturation of dendritic cells (DCs). , and the activation of effector T lymphocytes and the low infiltration efficiency of tumors lead to unsatisfactory therapeutic effects of cancer vaccines. Therefore, a size-variable antigen carrier with positive surface charge activat...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K31/4439A61K31/4188A61K9/107A61K47/34A61P35/00B82Y5/00B82Y40/00
CPCA61K39/0011A61K31/4439A61K31/4188A61K9/1075A61K47/34A61P35/00B82Y5/00B82Y40/00A61K2300/00
Inventor 杨红谢小雪刘贻尧李亭亭吴春惠李顺秦翔
Owner UNIV OF ELECTRONIC SCI & TECH OF CHINA
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