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Drug-loaded nano-micelle preparation as well as preparation method and application thereof

A drug-loaded nano and micelle technology, applied in the field of biomedicine, to achieve high affinity, improve solubility and biological stability, and improve the effect of binding ability

Inactive Publication Date: 2021-08-20
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no use of peptide drugs and their micellar preparations for the treatment of pancreatic cancer.

Method used

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  • Drug-loaded nano-micelle preparation as well as preparation method and application thereof
  • Drug-loaded nano-micelle preparation as well as preparation method and application thereof
  • Drug-loaded nano-micelle preparation as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] This example is used to illustrate the preparation method of the polypeptide drug of the present invention and its micelle preparation.

[0085] (1) Preparation method of PEGylated phospholipid molecular solution: dissolve PEGylated phospholipid powder in appropriate amount of ultrapure water, and obtain 1 mM PEGylated phospholipid molecular solution after complete dissolution.

[0086] (2) E5 polypeptide molecular solution: dissolve the powder of E5 polypeptide in an appropriate amount of ultrapure water, and sonicate for 5 minutes to obtain a 500 μM E5 polypeptide molecular solution.

[0087] (3) Mix the two molecular solutions in step (1) and step (2), incubate in a water bath, and let stand to obtain a drug-loaded nano-micelle solution of the polypeptide.

[0088] (4) Sterilize the drug-loaded nanomicelle solution obtained in step (3) after standing still, and then filter with a 0.22 μm filter membrane.

Embodiment 2

[0089] Example 2: Determination of the expression level of CXCR4 on the cell surface

[0090] When measuring the expression of CXCR4 on the cell surface by flow cytometry, divide the cells into 1.5mL centrifuge tubes, about 500,000 cells per tube, and incubate the antibody (primary antibody, mouse anti-human) of chemokine receptor CXCR4 in the experimental group ), pipette the cells evenly, incubate at room temperature for 1 hour, then centrifuge the cell suspension, discard the supernatant, resuspend with PBS, and centrifuge again, repeat this three times, wash away non-specifically bound antibodies, and then add 200 μL to prepare AF488-labeled goat anti-mouse secondary antibody solution, the same secondary antibody solution was also added to the control group, the cells were blown evenly, and incubated together at room temperature for 30min, then the cell suspension was centrifuged, the supernatant was discarded, and resuspended with PBS , centrifuge again, and repeat this...

Embodiment 3

[0091] Embodiment 3: E5 polypeptide and CXCR4 affinity determination

[0092] Flow cytometry was used to detect the binding ability of the E5 polypeptide and different chemokine receptor CXCR4 expression cell lines. Divide the cells into 1.5mL centrifuge tubes, about 500,000 cells per tube, discard the supernatant after centrifugation. Prepare FITC-E5 peptide solutions with concentrations of 2, 4, 6, 8, and 10 μM, respectively, and add 50 μL to the centrifuge tubes where the cells were collected. In the control group, no peptide was added, and 200 μL of PBS buffer was added, and gently blown with a pipette Incubate at 37°C for 2 hours until the cells are suspended evenly. Add an appropriate amount of PBS buffer to resuspend the incubated cells, centrifuge, discard the supernatant, resuspend the cells with an appropriate amount of PBS buffer, centrifuge again, repeat 3 times, wash away non-specifically bound polypeptides, and finally wash with 200 μL of PBS The buffer was r...

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Abstract

The invention provides a drug-loaded nano-micelle preparation as well as a preparation method and application thereof. The invention also provides a preparation method of the polypeptide drug and the nano-micelle preparation thereof. Wherein the drug-loaded nano-micelle preparation is formed by assembling poly(ethylene glycol)-phosphatidylethanolamine and a cancer therapeutic polypeptide. The polypeptide drug can be specifically combined with a chemokine receptor CXCR4, and migration of pancreatic cancer cells induced by a chemotactic axis CXCL12 / CXCR4 is inhibited. The polypeptide drug and the micelle preparation thereof provide a feasible method and technology for treatment of pancreatic cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a drug-loaded nano-micelle preparation, a preparation method and application thereof. Background technique [0002] Pancreatic cancer is the most malignant tumor of the digestive tract, with a very high lethality rate, and its five-year survival rate is less than 10%. Chemotherapy is the most effective treatment for pancreatic cancer except surgery. However, even the first-line chemotherapy drug gemcitabine is far less effective than clinical expectations. [0003] Chemokine CXCL12 and its receptor CXCR4 play an important role in the development of many cancers. CXCR4 is highly expressed in pancreatic cancer, breast cancer, non-small cell lung cancer, and esophageal cancer, and is associated with lymph node metastasis and poor prognosis. Among them, CXCR4 is highly expressed in pancreatic cancer tissues, and is closely related to the differentiation, metastasis, growth an...

Claims

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Application Information

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IPC IPC(8): A61K9/107A61K9/08A61K9/19A61K47/60A61K38/16A61P35/00A61P35/04
CPCA61K9/08A61K9/1075A61K9/19A61K38/16A61K47/605A61P35/00A61P35/04
Inventor 杨延莲李诗琪方小翠王琛
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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