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Liposome co-loaded with CD73 antibody and adriamycin, and preparation method and application of liposome

A doxorubicin and liposome technology, applied in the field of liposomes co-loaded with CD73 antibody and doxorubicin and their preparation, can solve problems such as drug resistance, reduce drug resistance and improve immune depletion state , the effect of promoting accumulation

Active Publication Date: 2021-09-28
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Doxorubicin (DOX) is an anthracycline drug with a broad anti-tumor spectrum, strong anti-tumor effect, and definite curative effect. It can not only kill tumor cells, but also promote immunogenic cell death (ICD) of tumor cells. Adriamycin, a chemotherapeutic drug used clinically, can cause serious side effects, such as hair loss, bone marrow suppression, and cardiotoxicity, especially cardiotoxicity is the most serious side effect of this type of drug, which limits its clinical application. And long-term doxorubicin chemotherapy can lead to drug resistance

Method used

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  • Liposome co-loaded with CD73 antibody and adriamycin, and preparation method and application of liposome
  • Liposome co-loaded with CD73 antibody and adriamycin, and preparation method and application of liposome
  • Liposome co-loaded with CD73 antibody and adriamycin, and preparation method and application of liposome

Examples

Experimental program
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Effect test

preparation example 1

[0035] Preparation of immunochemotherapeutic liposomes (CDL) co-loaded with CD73 antibody and DOX

[0036] The CD73 antibody used in the examples of the present invention is a commercial product purchased from BioXcell Biotechnology Co., Ltd., USA.

[0037] 1. Preparation of DOX-loaded liposomes (DOX Lip)

[0038] First, dissolve the appropriate concentration of DOPC:DOPE:CHEMS= molar ratio 2:6:1 in 3mL of chloroform, rotary evaporate, the temperature of the water bath is 22°C, and the rotation speed is 80rpm / min, finally forming a transparent spiral After the lipid film, dry thoroughly to remove the solvent.

[0039] 2. Add an ammonium sulfate solution (250mmoL, pH=5.5) properly preheated to 40°C for ultrasonication in a water bath, and repeat extrusion several times with a micro liposome extruder. The obtained solution was put into a treated dialysis bag (3000 Da), stirred and dialyzed in PBS solution for 4 hours at a constant speed, and the dialysate was changed every 2 h...

Embodiment 1

[0042] Example 1 Research on CDL Release Behavior in Vitro

[0043] Experimental method: Put the same volume of free doxorubicin (free DOX) and CDL preparation (preparation example 1) in the dialysis bag, put all the samples in PBS and shake at constant temperature to simulate the in vivo environment, 37°C, 100 r / min, the sample solution was sampled at different time points (0.25, 0.5, 1, 2, 4, 6, 8, 10, 12, 24, 48, 72 h, 96 h), and the PBS medium with the same pH was added again. Pass the sample solution taken out through a 0.22 μm organic filter membrane, use HPLC to detect and record its peak area, and analyze the release behavior of the sample in vitro. The results of CDL release behavior in vitro are as follows: figure 1 shown.

[0044] Depend on figure 1 It can be seen that the release rate of doxorubicin in CDL is significantly reduced. At pH=7.4, the cumulative release rate of the preparation at 72h was 23.15±0.58%, showing a significant sustained release effect. ...

Embodiment 2

[0045] Example 2 The Killing Effect of CDL on Triple Negative Breast Cancer Cells

[0046] Cell killing experiment: triple-negative breast cancer cells (4T1 cells) were mixed with 4×10 4 cells / mL, 200 μL / well were inoculated in a 96-well plate, and cultured overnight to make them adhere to the wall. The experimental groups were grouped as follows: blank liposomes, doxorubicin liposomes (DOX Lip), liposomes (CDL) co-loaded with doxorubicin and CD73 antibody, and the control group was added with complete medium, which were converted into DOX concentrations ( 0.002, 0.02, 0.2, 2, 20μg / mL) were added to the corresponding 96-well plate; the blank liposome group was added to the corresponding well at the concentration of (0.02, 0.2, 2, 20, 200μg / mL), and 100μL was added to each well drug volume. Place it in the incubator for 24h / 48h after the operation is completed. Add the prepared CCK-8 solution (10 μL / well) to each sample well and place it in the incubator for 1 hour. The max...

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Abstract

The invention provides a liposome co-loaded with CD73 antibody and adriamycin, and a preparation method and application of the liposome. A lipid core coated with the adriamycin is arranged in the liposome co-loaded with the CD73 antibody and the adriamycin, the outside of the lipid core is modified with the CD73 antibody, and the CD73 antibody is coupled to a phospholipid PEG active ester micelle and then is modified on the surface of the lipid core. The lipidosome co-loaded with the CD73 antibody and the adriamycin reduces the drug resistance and the toxic and side effects of the adriamycin in treatment of the triple negative breast cancer, improves in-vivo biological distribution of the adriamycin, improves bioavailability, and turns an immune microenvironment in a triple negative breast cancer tumor, so that the triple negative breast cancer tumor is converted into a thermal tumor, and a relatively good treatment effect for the triple negative breast cancer is realized.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a liposome co-carrying CD73 antibody and doxorubicin, a preparation method and application thereof. Background technique [0002] Breast cancer is the most common malignant tumor in women all over the world, and its morbidity and mortality rate ranks first among various cancers. Triple negative breast cancer (TNBC) is one of the subtypes. Its histopathological grade is poor, the degree of malignancy is high, and the tumor tissue grows rapidly. Once it develops, it progresses rapidly, and the tumor is prone to early recurrence The biggest feature of metastasis is its strong invasiveness. Simply put, it is particularly easy to transfer to other tissues of the body through blood or lymph, mainly manifested as visceral metastasis (lung metastasis and liver metastasis) and brain metastasis. And its clinical prognosis is poor, it is not sensitive to various treatment options, and the treatm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61K31/704A61K9/127A61K47/60A61P35/00
CPCA61K39/39558A61K31/704A61K9/1271A61K47/60A61P35/00A61K2300/00
Inventor 郭红梅祁同同林贵梅
Owner SHANDONG UNIV
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