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Nano particle carrier having tumor and tumor-associated macrophage targeting multiple functions and preparation thereof

A functional nano, tumor-related technology, applied in the field of multiple functional nanoparticle carriers and their preparation, can solve the problem that chemotherapy drugs cannot specifically act on tumor cells, cannot detect early tumors in time, and cannot accurately locate and detect tumor metastasis Stove and other issues

Inactive Publication Date: 2013-11-27
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problems that early tumors cannot be detected in time, tumor metastases cannot be accurately located and found, and chemotherapy drugs cannot specifically act on tumor cells, and a new tumor and tumor-associated macrophage Cell targeting multifunctional liposome nanoparticle carrier and preparation method

Method used

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  • Nano particle carrier having tumor and tumor-associated macrophage targeting multiple functions and preparation thereof
  • Nano particle carrier having tumor and tumor-associated macrophage targeting multiple functions and preparation thereof
  • Nano particle carrier having tumor and tumor-associated macrophage targeting multiple functions and preparation thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0061] Preparation of multifunctional liposome nanoparticle carrier with tumor and tumor-associated macrophage targeting (see figure 1 )

[0062] (1) mix the following substances;

[0063] ●DOPE 3mM

[0064] ●DOPC 3mM

[0065] ●Cholesterol 3mM

[0066] ●DOPE-PEG 2000 0.45mM

[0067] ●DSPE-PEG 2000-NH2 0.45mM

[0068] ●Legumain-TAT 0.51mM

[0069] (2) Dissolve the above substances in 50ml of organic solvents such as methanol, ethanol, or chloroform, and place the mixed solution at 4°C overnight;

[0070] (3) Rotary evaporation of organic solvent, 40°C, 10 minutes, continuous nitrogen drying, 1 hour;

[0071] (4) Add 1ml of 300mM ammonium hydrogen phosphate solution and hydrate for 1-2 hours;

[0072] (5) Mix with a vortex instrument;

[0073] ⑹ water bath ultrasound 20min, probe ultrasound 5min;

[0074] (7) Unify the particle size of nanoparticles into particles of about 100nm by extrusion, and store at 4°C;

[0075] (8) Use the ammonium hydrogen phosphate concentra...

Embodiment 2

[0079] Analysis of the uptake ability of tumor cells to multifunctional nanoparticle carriers with tumor and tumor-associated macrophage targeting in vitro

[0080] (1) Spread 4T1 cells on the coverslip of a 24-well plate at a density of 2×105 cells / well for laser confocal microscope detection;

[0081] (2) Spread 6-well plates with 4T1 cells at a density of 1.5×10 6 Cells / well, flow cytometry detection;

[0082] (3) Use CoCl 2 Stimulate the 4T1 cells plated in steps (1) and (2) to induce a hypoxic microenvironment;

[0083] (4) 200 microliters of nanoparticle carriers were added after 24 hours of hypoxia induction, and the in vitro uptake ability of 4T1 cells was tested;

[0084] (5) After adding the nanoparticle carrier for 2 hours, remove the medium and wash the cells with PBS;

[0085] (6) The cells used in step (1) for laser confocal microscope detection are processed according to the following steps;

[0086] ●Fix with pre-cooled acetone at -20°C for 20 minutes;

...

Embodiment 3

[0101] Biodistribution of Nanoparticle Carriers Targeting Tumor and Tumor-Associated Macrophages in BALB / c Mice

[0102] (1) Inject 1X10 orthotopically under the No. 4 fat pad of 6-8 week-old female BALB / c mice 5 After one week, firefly luciferase-labeled 4T1 mouse breast cancer tumor cells will form an orthotopic mouse breast cancer tumor in the No. 4 fat pad of the mouse, and establish a BALB / c mouse orthotopic breast cancer model;

[0103] (2) According to the method in Example 1, multiple functional nanoparticle carriers with tumor and tumor-associated macrophage targeting properties were prepared;

[0104] (3) Treat BALB / c mice with tumor-bearing mice by tail vein injection, and divide them into 6 groups. The first group is the negative control PBS group; the second group is the non-targeting simple liposome carrier packaging Doxorubicin group; the third group was the group of Doxorubicin packaged by Legumain-targeted liposome carrier; the fourth group was the group of D...

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Abstract

The invention relates to a nano particle carrier having tumor and tumor-associated macrophage targeting multiple functions, which can be used for tumor therapy and in vivo imaging and positioning. The carrier is a liposome nano particle mediated by a tumor focus induction-activated cell-penetrating peptide, can be specifically enriched on the tumor focus through a circulating system after being subjected to intravenous injection, and simultaneously has the functions of molecule imaging and carrying multiple biomacromolecules and medicines. Concretely, by using a TAT cell-penetrating peptide of closed active center amino acid of a Legumain specificity-identified enzyme digestion substrate to connect PEG-modified liposome nano particles, after the circulating system reaches the tumor focus position, Legumain expressed by tumor and tumor-associated macrophage can separate the enzyme digestion substrate from the TAT active center, thereby realizing the enrichment on the tumor focus. The nano particle can pack various biomacromolecules or medicines, and has the functions of killing tumor, in vivo tumor tracing and performing early prevention on tumor.

Description

technical field [0001] The invention belongs to the technical field of chemical materials, and relates to a novel multifunctional nanoparticle carrier with tumor and tumor-associated macrophage targeting and a preparation method thereof. The structure of the targeting group of the liposome nanoparticle as well as its linking method and function are mainly clarified. Background technique [0002] At present, malignant tumors have become the number one cause of unnatural death in the world, seriously threatening the physical and mental health of human beings. Under today's technical conditions, early tumors cannot be detected in time, resulting in missing the best opportunity for treatment, and the inability to accurately locate and find tumor metastases also reduces the survival rate of patients with malignant tumors. At present, chemotherapy is often used for treatment. Most chemotherapy drugs cannot specifically act on tumor cells. While killing tumor cells, they also have...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/42A61K9/14A61P35/00
Inventor 刘赜谭小月熊敏刘淑吕丹向荣
Owner NANKAI UNIV
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