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Biological membrane-wrapped multi-mode tumor contrast agent as well as preparation method and application thereof

A biofilm and multi-modal technology, applied in the direction of general/multifunctional contrast agents, antineoplastic drugs, preparations for in vivo experiments, etc., can solve the problems of lack of early diagnosis methods and difficulties in early diagnosis of deep tumors, and achieve improved Tissue penetration and the ability to be taken up by target cells, enhance tumor targeting, and enhance the effect of contrast

Inactive Publication Date: 2018-09-18
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally speaking, there is still a lack of ideal and specific early diagnosis methods, especially for early diagnosis of deep tumors.

Method used

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  • Biological membrane-wrapped multi-mode tumor contrast agent as well as preparation method and application thereof
  • Biological membrane-wrapped multi-mode tumor contrast agent as well as preparation method and application thereof
  • Biological membrane-wrapped multi-mode tumor contrast agent as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Preparation of melanoma targeting contrast agent

[0037] (1) Preparation of superparamagnetic ferritin nanocages (HFn)

[0038] The positive strains expressing superparamagnetic ferritin nanocages were constructed by Hubei Bios Biotechnology Co., Ltd. The company provides services such as plasmid construction, and the cells are also derived from this company.

[0039] Extract RNA from human skeletal muscle cells, reverse transcribe it into cDNA as a template; amplify human FTH1 gene, PCR primers are as follows (black underline is restriction site)

[0040] F: 5'A GTC GCC CAT ATG ACG ACC GCG TCC 3'(Nde I)

[0041] R:5'GCC GGA TCC TTA GCT TTC ATT ATCA C 3’(Bam HI)

[0042]After the PCR product was purified and recovered, it was ligated to the pET-30a(+) carrier by double enzyme digestion, and the product was transformed into XL2-Blue competent cells, screened on a Kan+ resistance plate, and colonies were picked and identified before being sent for seque...

Embodiment 2

[0053] Example 2 Preparation of Glioma Targeted Contrast Agent

[0054] (1) Preparation of ferritin nanocages of superparamagnetic ferric oxide:

[0055] Preparation of ferritin nanocages of superparamagnetic ferroferric oxide: Add 8.0mL of degassed, 100mM NaCl solution to the reaction flask (argon protection, strictly anaerobic operation), followed by 2.0mg, 3.9nmol superparamagnetic iron Protein nanocages (HFn1mg / mL) were added, the temperature was controlled at 65°C, pH=8.5 12.5mM, 1566 μL of degassed ferrous ammonium sulfate, the stoichiometric ratio of H 2 o 2 :Fe 2+ =1:3 Prepare 4.17mM, 1566μL degassed hydrogen peroxide solution, 25mM, 1566μL NaOH solution for neutralizing the H generated during the reaction + To maintain pH = 8.5; the three were added at a constant rate of 31.3 μL / min at the same time, and the theoretical load was 5000 Fe; after all were added, the reaction continued for 5 minutes; after the reaction was completed, 200 μL (300 mM) sodium citrate was ...

Embodiment 3

[0062] The stability test of embodiment 3 nanoparticles

[0063] The superparamagnetic ferritin nanocages with theoretical loadings of 1000Fe, 3000Fe, and 5000Fe prepared in Example 1 were used respectively, and the whole blood of human type O blood was processed to obtain 10 mg of biofilm, which was dissolved in 1 mL of water and 100 μL of normal saline (including erythrocytes). Membrane, white blood cell membrane and platelet membrane), when the red blood cell expands and hemolyzes in hypotonicity, many membrane pores with a diameter of 20-50nm will be formed briefly on the cell membrane, add M-HFn, incubate for 30min, centrifuge, collect the precipitate, and finally obtain the theory of different loads RBC ghost suspension loaded with M-HFn. Next, the erythrocyte ghost was repeatedly extruded by 400nm, 200nm, 100nm or 50nm liposome preparation apparatus (Avanti mini extruder) to prepare RBC-1000Fe, RBC-3000Fe and RBC-5000Fe aqueous solutions respectively.

[0064] Take 1 m...

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Abstract

The invention provides a biological membrane-wrapped multi-mode tumor contrast agent as well as a preparation method and application thereof. The biological membrane-wrapped multi-mode tumor contrastagent comprises a biological membrane, a ferritin nanometer cage (M-HFn) of superparamagnetic ferroferric oxide and a fluorescent dye, wherein the ferritin nanometer cage (M-HFn) of the superparamagnetic ferroferric oxide and the fluorescent dye are wrapped with the biological membrane and are connected with each other to form a fluorescent dye-M-HFn compound, a bionic nanometer biological membrane vector delivery system in vivo is constructed, in vivo long circulation can be realized, targeting to a tumor is strengthened, non-specific accumulation is reduced, damages on normal tissues are reduced, and the contrast effect is strengthened at the same time. The invention also provides a preparation method of the biological membrane-wrapped multi-mode tumor contrast agent, and the preparationmethod has important application prospect and study value in the field of clinic oncology early diagnosis and treatment and the like.

Description

technical field [0001] The invention belongs to the field of nuclear magnetic resonance contrast agents, in particular to a biofilm-wrapped multimodal tumor contrast agent and its preparation method and application. Background technique [0002] Cancer is one of the biggest killers threatening human life around the world, and it is a major challenge in the field of medical research. Magnetic resonance imaging has become an important basis for the diagnosis of primary tumors and metastases. The specificity and sensitivity of current tumor detection methods are low, and it is impossible to achieve specific recognition of tumor cells and controllable delivery of drugs at the same time. With the continuous development of nanotechnology, nanofluorescent probes have been widely used in chemistry, biology, medicine and other fields, especially in the diagnosis, imaging and treatment of tumors. The detection and imaging of multiple tumor markers with high sensitivity and high sele...

Claims

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Application Information

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IPC IPC(8): A61K49/18A61K49/00A61K9/50A61K47/46A61P35/00
CPCA61K9/5068A61K49/0002A61K49/0021A61K49/0097A61K49/1866A61K49/1896A61P35/00
Inventor 戴箭宋镕光阮淼亮薛巍
Owner JINAN UNIVERSITY
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