Double-target tumor nanoliposome and preparation method thereof

A nano-liposome, dual-targeting technology, applied in the field of medicine, to achieve the effect of excellent specific binding force and small molecular weight

Inactive Publication Date: 2011-09-21
SHANGHAI PULMONARY HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Due to the heterogeneity of tumor vascular endothelial cells under the influence of the microenvironment, different types of tumors express different types and quantities of antigens and receptors, and the affinity of the same ligand or antibody to different types of tumor vascular endothelial cells is also different. There are large differences, and no target molecule has been found that can recognize a variety of tumors with high specificity and high affinity

Method used

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  • Double-target tumor nanoliposome and preparation method thereof
  • Double-target tumor nanoliposome and preparation method thereof
  • Double-target tumor nanoliposome and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Synthesis steps of single-target and dual-target tumor polypeptides

[0040] The dual-targeting peptide was synthesized by 9-fluorenylmethyloxycarbonyl (FMOC) solid-phase synthesis method, purified by high performance liquid chromatography (HPLC), and identified by mass spectrometry. Concrete synthetic steps are as follows:

[0041] 1) Resin swelling: put FMOC-AA-Wang-Resin resin into the reaction tube, add dimethylformamide (N, N-dimethylformamide, DMF) (15ml / g) for 30min; deprotection: discard DMF, Add 20% piperidine DMF solution (15ml / g) for 5min, then add 20% piperidine DMF solution (15ml / g) for 15min after discarding;

[0042] 2) Detection: Take out the piperidine solution, take more than a dozen resins, wash them three times with ethanol, add ninhydrin, potassium cyanide, and phenol solution one drop each, heat at 105°C-110°C for 5 minutes, and turn dark blue as a positive reaction.

[0043] 3) Washing: DMF (10ml / g) twice, methanol (10ml / g) twice, DMF...

Embodiment 2

[0052] Example 2: Observation of the binding force between the dual-targeting polypeptide and cells by flow cytometry

[0053] A549 cells and HUVEC were cultured in 6-well plates, and when the cells>90% of the culture plate, fluorescein isothiocyanate (FITC)-labeled single-target polypeptide (FITC- ATWLPPR, FITC-ARYC RGD CFDG) and dual targeting peptides (FITC-ARYC RGD CFD ATWLPPR ) was diluted with serum-free DMEM culture solution to a final concentration of 2 μmol / L and added to each well, incubated in a 5% constant temperature incubator at 37°C for 3 hours, washed with PBS buffer (pH 7.4), and digested with 0.25% trypsin for each well , collect the cells, wash once with 0.5ml Coulter Isoton III diluent, add 0.5ml Coulter Isoton III diluent, vortex and mix well, then use the F1 channel to detect the relative fluorescence intensity of the cells on the machine (results see figure 2 ).

[0054] The results can be seen in FITC-ARYC RGD CFDG and FITC-ARYC RGD CFD ATWL ...

Embodiment 3

[0055] Example 3: Observation of the binding ability of dual-targeting polypeptides and cells with a fluorescent microplate reader

[0056] A549 cells and HUVEC were seeded in 24-well plates. When the cells were >90% of the culture plate, the FITC-labeled polypeptide was diluted with serum-free DMEM culture solution to a final concentration of 5 μmol / L and added to each well, and cultured at 37°C and 5% constant temperature. Incubate in the box for 3 hours, wash with PBS buffer (PH7.4) for 3 times, add 200 μL of cell lysate to each well to lyse the cells, take 100 μL and add it to a fluorescent microplate plate, use a fluorescent microplate reader to detect the fluorescence intensity, the absorption wavelength is 485 / 20nm, The emission wavelength is 528 / 20nm, see the attached results image 3 .

[0057] The results showed that FITC-ARYC RGD CFDG, FITC-ATWLPPR and FITC-ARYC RGD CFD ATWLPPR The relative fluorescence intensity was significantly higher than that of FITC-label...

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Abstract

The invention belongs to the field of biomedicines, and particularly discloses a double-target tumor nanoliposome containing ARYCRGD CFDATWLPPR polypeptide; and the invention further discloses a preparation method for the double-target tumor nanoliposome, and applications of the nanoliposome in preparing antitumor medicaments.

Description

technical field [0001] The invention belongs to the field of medicine, and more specifically, the invention discloses a tumor-targeting nano liposome and a preparation method thereof. Background technique [0002] In recent years, the morbidity and mortality of malignant tumors have shown an obvious upward trend, and have become a major disease that threatens human health and life. According to a report by the International Agency for Research on Cancer, the number of cancer cases worldwide doubled between 1975 and 2000, with approximately 12 million new cases and more than 7 million deaths each year. In 2010, cancer will become the leading cause of death in the world. In 2030, the number of cancer patients will triple, and the number of new cases will increase to 20-26 million. [0003] Chemotherapy is one of the main methods in the comprehensive treatment of malignant tumors. A reasonable chemotherapy strategy can significantly prolong the survival time of cancer patients...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/127A61K47/48A61K47/42A61K49/14C07K7/08A61P35/00
Inventor 周彩存孟淑燕粟波李玮宋胤周蔚
Owner SHANGHAI PULMONARY HOSPITAL
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