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Golgi apparatus and genetic engineering exosome hybrid membrane coated retinoic acid in-situ spray hydrogel vaccine, and preparation method and application thereof

A technology of genetic engineering and Golgi apparatus, applied in the suppression and/or treatment of postoperative recurrence of melanoma, in the field of retinoic acid in situ spray hydrogel vaccine, can solve the problem of low response rate of immunotherapy, recurrence of patients, etc. problem, to achieve the effect of prolonging the survival time and reducing the recurrence rate

Active Publication Date: 2021-07-02
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the response rate of patients to immunotherapy is still very low, and the vast majority of patients will relapse within a few months

Method used

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  • Golgi apparatus and genetic engineering exosome hybrid membrane coated retinoic acid in-situ spray hydrogel vaccine, and preparation method and application thereof
  • Golgi apparatus and genetic engineering exosome hybrid membrane coated retinoic acid in-situ spray hydrogel vaccine, and preparation method and application thereof
  • Golgi apparatus and genetic engineering exosome hybrid membrane coated retinoic acid in-situ spray hydrogel vaccine, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] A method for preparing retinoic acid biomimetic nanoparticles (GENPs) coated with hybrid membranes of Golgi apparatus and genetically engineered exosomes, comprising the following steps:

[0127] The preparation details of GENPs, such as figure 1 shown.

[0128] Step 1: preparing nanoparticles of retinoic acid;

[0129] Retinoic acid-loaded PLGA nanoparticles were prepared by the oil-in-water (O / W) emulsification solvent evaporation method. Dissolve 60 mg of PLGA and 2 mg of retinoic acid in 4 mL of dichloromethane. The organic phase was transferred to 8 mL of 1.5% PVA solution. Then, the solution was sonicated at 400 W for 5 minutes in an ice bath. The emulsion was then shaken overnight at room temperature to eliminate the organic solvent and form uniformly dispersed nanoparticles. Subsequently, the particle suspension was centrifuged at 16000g for 20 minutes. The prepared nanoparticles were then washed three times with an equal volume of deionized water.

[013...

Embodiment 2

[0142] Prepare a dosage form, which is retinoic acid biomimetic nanoparticles coated with Golgi apparatus and genetically engineered exosome hybrid membrane, and loaded with PVAA / CS gel (aPD-L1@GENPs@Gel);

[0143] The method of Example 1 was used to prepare retinoic acid biomimetic nanoparticles GENPs coated with hybrid membranes of Golgi apparatus and genetically engineered exosomes; the obtained GENPs and PD-L1 monoclonal antibodies were loaded into PVAA and CS respectively to obtain in situ spray Hydrogel vaccine, which inhibits the secretion of exosome PD-L1, revitalizes lymph node immune cells to trigger immune memory and then activates the Golgi apparatus of the systemic immune response and the retinoic acid biomimetic hydrogel coated with the hybrid membrane of genetically engineered exosomes vaccine. Specifically:

[0144] Dry PVA powder (5.0 g) was added to a three-necked flask containing 45 mL DMSO. The mixture was heated to 90°C under magnetic stirring to complet...

Embodiment 3

[0147] Golgi-disrupting ability of retinoic acid biomimetic nanoparticles (GENPs) coated with hybrid membrane of genetically engineered exosomes and exosomal PD-L1 inhibitory ability

[0148] To assess the cell adhesion of GENPs, B16-F10 cells were incubated with fluorescently labeled nanoparticles for 1 h at 4 °C. Such as image 3 As shown in a and 3b, although both GENPs and ENPs showed high fluorescence signals on B16-F10 cells, GNPs showed weak adhesion to B16-F10 cells, indicating that EM coating promoted the adhesion of nanoparticles to tumor cells. affinity.

[0149] Confocal microscopy was used to study the cellular internalization of GENPs. B16-F10 cells were incubated with FITC-RA, PLGA-RA, GNPs and GENPs for 6 hours, respectively. Such as image 3 As shown in c, various intensities of fluorescence were observed in the cytoplasm. In B16-F10 cells, both GNPs and GENPs showed high intracellular fluorescence of FITC-RA ( image 3 d), which indicates that the enhan...

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Abstract

The invention discloses a Golgi apparatus and genetic engineering exosome hybrid membrane coated retinoic acid in-situ spray hydrogel vaccine as well as a preparation method and an application thereof, and belongs to the technical field of medicines. In particular to a retinoic acid in-situ spray hydrogel vaccine which inhibits exosome PD-L1 secretion, reproduces lymph node immune cells to trigger immune memory so as to activate systemic immune response and is coated with a Golgi apparatus and genetic engineering exosome hybrid membrane and an application of the retinoic acid in-situ spray hydrogel vaccine in inhibition and / or treatment of postoperative recurrence of melanoma.

Description

technical field [0001] The invention belongs to the technical field of medicines, and in particular relates to a hybrid membrane-coated vitamin of Golgi bodies and genetically engineered exosomes, which inhibits exosome PD-L1 secretion, revitalizes lymph node immune cells, triggers immune memory, and activates systemic immune response. Formic acid in situ sprayed hydrogel vaccine and its use in the suppression and / or therapeutic suppression of postoperative recurrence of melanoma. Background technique [0002] Local residual tumor and microinfiltration of circulating tumor cells (CTCs) tend to induce tumor recurrence. Immunotherapy can effectively suppress cancer recurrence and metastasis, among which PD-L1 monoclonal antibody shows great promise in treatment. However, the response rate of patients to immunotherapy is still very low, and most patients will relapse within a few months. This is because PD-L1 is also located on the surface of extracellular vesicles, enhances ...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/395A61K31/203A61K9/06A61K47/36A61K47/32A61K9/51A61K47/46A61P35/00
CPCA61K39/00119A61K39/39558A61K31/203A61K9/06A61K47/36A61K47/32A61K9/5161A61K9/5176A61P35/00A61K2039/876A61K2300/00
Inventor 孙进叶皓何仲贵
Owner SHENYANG PHARMA UNIVERSITY
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