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Immune-enhanced exosome hydrogel compound as well as preparation method and application thereof

A technology of immune enhancement and exosomes, which is applied in the direction of non-active ingredients of polymer compounds, drug combinations, and pharmaceutical formulations, and can solve the problems of low tumor adhesion, weak retention, and low targeting efficiency of ordinary hydrogels. To achieve the effect of overcoming uneven distribution of drugs, enhancing dispersion stability, and improving dispersion stability

Active Publication Date: 2021-06-11
AFFILIATED HOSPITAL OF JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Purpose of the invention: In order to solve the above technical problems, the present invention provides an immune-enhancing exosome hydrogel complex and its preparation method and application. The hydrogel complex is a brand-new chemotherapy + immune enhancer combination This solution can overcome the disadvantages of poor stability, low targeting efficiency, and weak retention of drug-loaded exosomes, as well as the shortcomings of ordinary hydrogels such as low tumor adhesion, non-injection, and non-responsive drug release, and improve the efficacy of drugs in tumor and tumor inflammatory environments. effective accumulation concentration, reduce systemic toxic and side effects, play a synergistic killing effect, and have significant therapeutic effects on various solid tumors

Method used

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  • Immune-enhanced exosome hydrogel compound as well as preparation method and application thereof
  • Immune-enhanced exosome hydrogel compound as well as preparation method and application thereof
  • Immune-enhanced exosome hydrogel compound as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0063] Example 1: Preparation and characterization of mannan peptide-hyaluronic acid hydrogel loaded with doxorubicin-M1 type RAW264.7 macrophage exosomes

[0064] (1) Under the condition of high glucose DMEM medium with 10% fetal bovine serum and 1% double antibody, in 5% CO 2 Mouse macrophages were cultured in a constant temperature incubator at 37°C. After the cells were completely attached to the wall and in a good growth state, 500 ng / ml LPS and 2 ng / mL IFN-γ were added and cultured for 24 hours to induce the formation of M1-polarized RAW264.7 macrophages. The original medium was sucked off, washed twice with PBS, and then serum-free high-glucose DMEM medium was added to culture for 48 hours. Collect 100mL of medium and use ultracentrifugation to separate M1-macrophage exosomes: centrifuge at 300g for 15min at 4°C, and take the supernatant; centrifuge at 3000g for 30min, take the supernatant; centrifuge at 10000g for 90min, and resuspend the pellet in 1×PBS Centrifuge at...

Embodiment 2

[0069] Example 2: Preparation and characterization of mannan peptide-hyaluronic acid hydrogel loaded with doxorubicin-M1 bone marrow-derived macrophage exosomes

[0070] (1) Take the leg bones of 4-6 week-old mice, wash out the bone marrow with PBS buffer, and centrifuge; use ACK to lyse red blood cells, and centrifuge in DMEM medium. Culture in 1.5ml six-well plate with high-glucose DMEM medium (10% fetal calf serum, M-CSF 20ng / ml) for 3 days, wash away floating cells, replace with new medium, and obtain adherent mouse bone marrow-derived macrophages . Under the condition of 10% fetal bovine serum high glucose DMEM medium and 1% double antibody, in 5% CO 2 Mouse macrophages were cultured in a constant temperature incubator at 37°C. After the cells were completely attached to the wall and in a good growth state, 500ng / ml LPS and 2ng / mL IFN-γ were added, and cultured for 24 hours to induce the formation of M1-polarized bone marrow-derived macrophages. The original medium was ...

Embodiment 3

[0073] Example 3: Dispersion stability, biosafety, degradation performance and drug release profile of exosome gel complexes

[0074] (1) The doxorubicin-M1 type RAW264.7 macrophage exosomes prepared in Example 1 were ultrasonically dispersed in a commercially available cross-linked hyaluronic acid gel (HA gel, Bloomage Biotechnology, product number TL100, Chinese patent ZL2015101091144), prepare DOX@M1-exos-HA gel. In order to investigate the dispersion stability of DOX@M1-exos in mannan peptide-hyaluronic acid hydrogel and ordinary cross-linked hyaluronic acid gel, the prepared DOX@M1-exos-HA-gel and DOX@M1 -exos-OHA / Man gel was placed in a refrigerator at 4°C, and the samples were observed on 0d and 7d respectively, such as Figure 9 As shown, DOX@M1-exos in ordinary cross-linked hyaluronic acid gels aggregated and precipitated after 7 days, while DOX@M1-exos in mannan peptide-hyaluronic acid hydrogels dispersed uniformly, presumably due to The mannose receptor on the sur...

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Abstract

The invention discloses an immune-enhanced exosome hydrogel compound as well as a preparation method and application thereof, the compound is prepared by the following steps: encapsulating a drug by an M1 type macrophage exosome to form a medicine-M1 type macrophage exosome, and then forming hydrogel through chemical crosslinking of mannatide and hyaluronic acid, and loading the medicine-M1 type macrophage exosome in the hydrogel to form the hydrogel medicine delivery system. The mannatide-hyaluronic acid hydrogel prepared by the invention not only has the advantages of acid-sensitive drug release, injectability, self-healing and the like of Schiff base type gel, but also has the effects of enhancing anti-tumor immunity, increasing exosome stability, tumor adhesiveness, biodegradability, biosafety and the like; and the drug-M1 type macrophage exosome can be slowly released after tumor local injection administration, and the drug-M1 type macrophage exosome can permeate into tumor and tumor inflammation areas in a targeted manner, so that tumor immunity is activated, and tumor is cooperatively killed.

Description

technical field [0001] The invention discloses an immune-enhancing exosome hydrogel complex, a preparation method and application thereof, and belongs to the technical field of hydrogel complexes. Background technique [0002] Malignant tumors seriously threaten human health, and existing treatment options such as surgery, chemotherapy, radiotherapy, targeted therapy, and immunotherapy all have certain limitations. Although the combination therapy of chemotherapy and immunization has been commonly used in tumor therapy, it mostly adopts a systemic drug regimen, which has disadvantages such as low tumor selectivity, weak immune response efficiency, and high toxicity and side effects, which limit its clinical application. [0003] In recent years, local drug delivery to tumors, as an effective improvement scheme for systemic treatment, has become one of the research and development hotspots in the field of anti-tumor. Exosomes are nanovesicles secreted by cells with a bilayer...

Claims

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Application Information

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IPC IPC(8): A61K35/15A61K9/06A61K31/12A61K31/122A61K31/185A61K31/337A61K31/352A61K31/575A61K31/704A61K45/06A61K47/36A61K47/42A61P35/00A61P37/04
CPCA61K9/06A61K31/12A61K31/122A61K31/185A61K31/337A61K31/352A61K31/575A61K31/704A61K35/15A61K45/06A61K47/36A61K47/42A61P35/00A61P37/04A61K2300/00
Inventor 宋金方倪江叶琳岚马昂李霞丁永娟
Owner AFFILIATED HOSPITAL OF JIANGNAN UNIV
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