Nano-drug carrier loading anticancer drugs, and preparation method and application of nano-drug carrier

A nano-drug carrier, anti-tumor drug technology, applied in the field of immunology and biomedicine, nanotechnology

Active Publication Date: 2019-03-26
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, excessive intake of drug carriers in the body will bring new side effects. Therefore, increasing the loading capacity of chemotherapy drugs of drug carriers is a problem that needs to be solved in the field of cancer treatment.

Method used

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  • Nano-drug carrier loading anticancer drugs, and preparation method and application of nano-drug carrier
  • Nano-drug carrier loading anticancer drugs, and preparation method and application of nano-drug carrier
  • Nano-drug carrier loading anticancer drugs, and preparation method and application of nano-drug carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1, Preparation and Characterization of Pyrococcus Furious Ferritin Nano-Drug Carrier Fn-Dox

[0033] Pyrococcus furiosus ferritin pfFn (Fn) is highly similar to ferritin from other species (human, mouse) in structure, and can self-assemble into a 24-mer spherical protein shell structure in a natural state, with an inner diameter of 8nm and an outer diameter of 12nm. As mentioned above, the structural characteristics of ferritin make it an obvious advantage as a drug carrier. According to the current research, the loading capacity of a single ferritin molecule to the first-line chemotherapy drug doxorubicin is about 20-40 molecules1-6, and the ferritin molecule modified by PAS polypeptide can load about 90 molecules of doxorubicin (Elisabetta Falvo. et al., Biomac, 2015). Our work found that using a relatively mild method of urea denaturation and renaturation to prepare ferritin doxorubicin nanoparticles, the doxorubicin loading capacity of Pyrococcus furiosus f...

Embodiment 2

[0044] Example 2. Preparation and characterization of liver cancer-targeted ferritin nano-drug carrier HccFn-Dox

[0045] By means of gene fusion, the SP94 liver cancer targeting peptide (SEQ ID NO: 2) is fused and expressed at the N-terminal of Pyrococcus furiosus ferritin (SEQ ID NO: 1) through the adapter protein (SEQ ID NO: 3), and we fused this The protein was named HccFn (SEQ ID NO: 4).

[0046] With the gradient urea dialysis method as described in Example 1, doxorubicin is loaded into the HccFn lumen ( figure 2 A). Superdex 200pg molecular sieves detect the ultraviolet absorption of A280nm and A485nm, and it can be seen that doxorubicin is successfully loaded into the inner cavity of HccFn ( figure 2 B). TEM ( figure 2 C) and dynamic light scattering ( figure 2 D) The analysis results also show that ferritin (Fn), HccFn and HccFn-Dox of Pyrococcus furiosus all maintain good uniformity and spherical shell structure. The results of native gel electrophoresis ( ...

Embodiment 3

[0048] Example 3. Liver cancer-targeting characterization of liver cancer-targeting ferritin nano-drug carrier HccFn-Dox in vivo and in vitro

[0049] 1. Cell targeting of HccFn-Dox

[0050] First, we use FITC to fluorescently label Fn and HccFn protein shells. The FITC fluorescent labeling method is as follows: Dissolve 1mg of Fn or HccFn protein shells in 0.1M carbonate buffer solution (pH9.0) (final protein concentration 1mg / ml), add 50 microliters of FITC solution (1mg / ml, dissolved in DMSO), and mix overnight at 4°C in the dark. Centrifuge at 12,000 g for 10 min to take the supernatant, and replace the PBS buffer with an ultrafiltration concentrator until the filtrate is colorless. FITC concentration was determined by UV absorbance (Nanodrop 2000).

[0051] Mix 1 μg / ml FITC-Fn and FITC-HccFn with 100 μL hepatoma cell HepG2 (2.5×10 6 cells / mL) were co-incubated for 30 minutes, observed by laser confocal scanning microscope, HccFn specifically bound to the surface of He...

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Abstract

The invention relates to a nano-drug carrier loading anticancer drugs, and a preparation method and application of the nano-drug carrier. The nano-drug carrier is a physiological polymer of pyrococcusfuriosus ferritin and a derivative thereof, wherein the derivative comprises fusion proteins modified based on the pyrococcus furiosus ferritin, and ferritin modified based on the amino acid sequenceof a protein shell inner cavity of the pyrococcus furiosus ferritin; and the anticancer drugs include but are not limited to small molecule drugs, oligonucleotides and functional peptide fragments.

Description

technical field [0001] The invention belongs to the technical fields of nanotechnology, immunology and biomedicine. Specifically, the present invention relates to a nano drug carrier loaded with antitumor drugs, its preparation method and application. Background technique [0002] Cancer has become one of the major diseases that are increasingly common and seriously threaten human life and quality of life. As the aging population continues to increase, the incidence of cancer is expected to continue to grow. Currently, there are three main methods of treating cancer: surgery, radiotherapy and chemotherapy. The choice of cancer treatment depends on the type, location, and spread of the cancer. Chemotherapy is a treatment method that uses chemical drugs to kill tumor cells and inhibit tumor growth. It is a systemic treatment that has therapeutic effects on primary tumors, metastases and subclinical metastases. For cancers that cannot be treated with surgery and radiation, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/42A61K45/00A61P35/00C07K14/195C12N15/70C12N15/31
CPCA61K9/5169A61K45/00A61P35/00C07K14/195C12N15/70
Inventor 阎锡蕴范克龙江冰
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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