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Fusion membrane-coated bionic nanoemulsion as well as preparation method and application thereof

A technology of biomimetic nanometer and fusion membrane, which is applied in the field of biomimetic nanoemulsion and its preparation, can solve the problems of low clinical efficiency of immune checkpoint inhibitors, killing normal tissues, and limited targeting ability of tumor sites, etc., achieving good application prospects, Easy to operate and improve the effect of curative effect

Pending Publication Date: 2021-09-17
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of the above-mentioned deficiencies in the prior art, the purpose of the present invention is to provide a fusion membrane-wrapped biomimetic nanoemulsion and its preparation method and application, aiming to solve the low clinical efficiency of existing immune checkpoint inhibitors and the targeting ability of tumor sites Limited, photothermal therapy has the problem of killing normal tissues

Method used

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  • Fusion membrane-coated bionic nanoemulsion as well as preparation method and application thereof
  • Fusion membrane-coated bionic nanoemulsion as well as preparation method and application thereof
  • Fusion membrane-coated bionic nanoemulsion as well as preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0088] In this example, the preparation steps of the biomimetic nanoemulsion wrapped in the fusion membrane (lactate oxidase-albumin-perfluorotributylamine wrapped in the fusion membrane, denoted as PHL@PSHM) are as follows:

[0089] Mix human serum albumin and water, the concentration of the mixed solution is 20 mg / mL, a total of 60 mg human serum albumin solution, then slowly add 300 μL perfluorotributylamine and stir to obtain a stirred solution. According to the volume ratio of 20:1, 300 μL of 2 mg / mL lactate oxidase dissolved in deionized water was added to obtain a mixed solution. The above mixed solution was ultrasonically treated with a probe (265W) for 6 minutes to obtain lactic acid oxidase-albumin-perfluorotributylamine biomimetic nanoemulsion.

[0090] Extract the cell membrane with high expression of PD-1: Digest HEK293T cells with trypsin, centrifuge at 1500rpm for 5 minutes to get the cells, wash with PBS 3 times, resuspend with HM buffer, and homogenize 100 wit...

Embodiment 2

[0096] Toxicity Evaluation of Starvation Treatment on 4T1 Tumor Cells

[0097] The standard MTT method was used to evaluate the effect of starvation treatment on the survival rate of 4T1 cells. 4T1 cells at 5×10 per well 3 Density seeded into 96-well plates and placed at 37°C, 5% CO 2 conditions for 24 hours. Next, suck out the old culture medium in the 96-well plate, and add the perfluorotributylamine-human serum albumin-lactate oxidase (PFTBA@ HSA / LOx, PHL) in DMEM medium. After continuing to cultivate for 24 hours, suck out the old medium in the 96-well plate, add 100 μL of medium solution containing 5 mg / mL MTT to each well, and continue to cultivate for 12 hours. Then detect the OD value of each well on a Bio-TelEL microplate reader (the detection wavelength is 490nm), and calculate the cell survival rate with the following formula. Cell viability (%)=(OD490 value of sample / OD490 value of blank)×100%, see experimental results figure 2 .

[0098] Such as figure 2...

Embodiment 3

[0100] Evaluation of the Effect of Salmonella Outer Membrane Vesicles Inducing Tumor Hemorrhage

[0101] Female Balb / c mice (3 weeks, 15-20g), subcutaneously inject 2×10 6 4T1 tumor cells were used to establish a mouse subcutaneous tumor model. When the tumor volume exceeds 100mm 3 1-3 mg / kg of biomimetic nanoemulsion PHL@PSHM wrapped in fusion membrane was injected into the tail vein, and photoacoustic imaging experiments were performed. At 0, 2, 4, 12, and 24 hours after the tail vein injection of the drug in the tumor-bearing mice, the changes of the 800nm ​​wavelength photoacoustic signal value in the tumor were observed by photoacoustic imaging and quantitatively analyzed. The experimental results are shown in image 3 .

[0102] From image 3 It can be seen from A and B that after administration, the photoacoustic signal at 800nm ​​in the tumor gradually increases with time after administration, and the photoacoustic signal at the tumor site in the 1mg / kg and 3mg / kg...

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Abstract

The invention discloses a fusion membrane-coated bionic nanoemulsion as well as a preparation method and application thereof. The fusion membrane-coated bionic nanoemulsion is prepared by the following steps of: providing a bionic nanoemulsion, wherein the bionic nanoemulsion comprises a shell and a core positioned in the shell, the shell is composed of lactic oxidase and human serum albumin, and the core is composed of a perfluorinated compound; providing a fusion membrane, wherein the fusion membrane is formed by fusing an engineered cell membrane and a bacterial outer membrane vesicle; and coating the bionic nanoemulsion with the fusion membrane to obtain the fusion membrane-coated bionic nanoemulsion. The fusion membrane-coated bionic nanoemulsion disclosed by the invention can simultaneously realize the synergistic treatment of tumor by combining photothermal therapy, starvation therapy and immunotherapy, thereby having a good application prospect in the field of tumor diagnosis and treatment. Meanwhile, the preparation process disclosed by the invention is simple and convenient to operate, does not need complex and expensive equipment, and is easy to realize industrial production.

Description

technical field [0001] The invention relates to the field of medical nanomaterials, in particular to a biomimetic nanoemulsion wrapped in a fusion membrane and a preparation method and application thereof. Background technique [0002] Immunotherapy, as an emerging cancer treatment, can destroy the microenvironment of tumor immune tolerance and induce tumor-specific immune responses, with far fewer side effects than traditional chemotherapy and radiotherapy. Over the past decade, immune checkpoint inhibitors have achieved remarkable clinical results in various types of tumors. However, the overall response rate for immune checkpoint antibodies is relatively low. In addition, these antibodies lack tumor-targeting ability and are distributed in various tissues throughout the body after injection, which may cause certain side effects (such as cytokine storm). The high cost of antibodies also imposes a heavy financial burden on cancer patients. The synergistic effect of immun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/51A61K9/107A61K9/51A61K47/46A61K41/00A61K47/42A61P35/00C12N1/20C12N5/071C12R1/19C12R1/01C12R1/42C12R1/46A61K31/02A61K31/131
CPCA61K9/5176A61K9/1075A61K41/0052A61K38/51A61K31/02A61K31/131A61K47/42A61P35/00C12N1/20C12N5/0686C12N2509/00C12N2509/10A61K2300/00Y02A50/30
Inventor 黄鹏张一帆廖云燕林静
Owner SHENZHEN UNIV
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