Novel tumor targeting contrast agent

A tumor targeting and contrast agent technology, applied in the field of biomedicine, can solve the problems of high dosage, low tumor targeting, toxic and side effects, etc.

Inactive Publication Date: 2012-05-23
杨静雯
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The contrast agents currently used in clinical practice mainly have problems such as high dosage, low tumor targeting, and certain toxic and side effects.

Method used

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  • Novel tumor targeting contrast agent
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Preparation of recombinant human heavy chain ferritin (Human H chain Ferritin, HFn)

[0035] The preparation method of recombinant human ferritin is mainly referenced (Masaki U., Michelle L.F., Mark A., Deborah A.W., Bridgid E.C., Susan B., Ann F.W., Larissa J., Mark J., Mark J.Y. and Trevor D ., J. AM. Chem. Soc. 2006, 128, 16626) described method. The specific implementation is as follows: design primers from normal human skeletal muscle cDNA and use PCR method to amplify human ferritin heavy chain gene. The upstream primer is: 5'-A GTC GCC CAT ATG ACG ACC GCG TCC; the downstream primer is: 5' ? GCC GGATCC TTA GCT TTC ATT ATC AC. The amplified target PCR fragment with a size around 550-600bp was recovered by NdeI / BamHI double enzyme digestion, and connected to the pET39b vector (Novagen) fragment that was also recovered by NdeI / BamHI double enzyme digestion. The HFn gene was identified by sequencing as correct. The resulting expression vector was tra...

Embodiment 2

[0039] Example 2. Preparation of human ferritin containing iron oxide core

[0040] This preparation process is always at N 2 environment. Add 8ml of degassed solution containing 100mM NaCl to N 2 Add 2ml of HFn with a concentration of 1mg / ml to a total of 2mg of samples (also dissolved in 100mM NaCl solution) into a protected airtight container, and titrate the pH of the system to 8.5 with 50mM NaOH while the circulating water maintains an ambient temperature of 65°C; Add 12.5 mM (NH4) at a molar ratio of 3000 iron atoms per clathrate cavity 2 Fe(SO4) 2 , and press H at the same time 2 o 2 :Fe(II) at a molar ratio of 1:3 was added with freshly prepared degassed H at a concentration of 4.17 mM 2 o 2 ; Fe(II) and H 2 o 2 Add to the reaction system at a fixed rate of 31.3ul / min, and at the same time use 50mM NaOH to titrate and stabilize the pH of the system at 8.5. After the reaction was completed, 200ul of 300mM sodium citrate was added to chelate excess free iron...

Embodiment 3

[0042] Example 3. MRI Scanning Imaging of Tumor-targeted Contrast Agent in Human Tumor-bearing Nude Mouse Model

[0043] Human lung cancer cell line SPCA1 was inoculated subcutaneously in nude mice (10 7 cells / mouse), and a total of 10 human-bearing lung cancer nude mice models were made. They were divided into two groups with 5 rats in each group, namely the observation group and the control group. When the tumors grow to 1-1.5cm in size, all nude mice were scanned by MRI, and the MRI signal intensity of the tumor lesions was determined to be 278. HFn ferritin, the tail vein of control group nude mice injected Fe with a diameter of 50nm at the ratio of 5mg Fe / kg body weight 3 o 4 Magnetic nanoparticles, MRI enhanced scan was performed half an hour later, and the MRI signal intensity of tumor lesions in the observation group was determined to be 451, while the MRI signal intensity of tumor lesions in the control group was 325.

[0044]Human liver cancer cell line hep3b ...

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Abstract

The invention provides a novel tumor targeting contrast agent, which is characterized in that an apoferritin cage cavity housing is assembled with heavy chain by human ferritin, and a pharmaceutically acceptable magnetic resonance imaging contrast reinforcing agent is coated in the apoferritin cage cavity, the formed compound is a novel MRI tumor imaging contrast agent possessing a tumor specific targeting function. The contrast agent of the invention has the characteristics of no toxicity or low toxicity, good tumor targeting, strong specificity, less usage amount, high in vivo stability and the like, and is especially suitable for magnetic resonance imaging contrast of tumor tissue like liver cancer tumor, melanin tumor, lung cancer tumor, breast cancer tumor, colon cancer/rectal cancer tumor, pancreas cancer tumor, stomach cancer tumor, ovary cancer, head and neck cancer, neuroglioma.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. Specifically, the invention provides a novel tumor targeting molecule, which is ferritin assembled from 24 protein subunit molecules, and the protein subunit molecule is human ferritin heavy and light chains. They can be assembled into a cage-like cavity structure of 24 subunits according to various molecular ratios, at least containing heavy chain subunits. The present invention also provides that the tumor targeting molecule can specifically recognize and bind to liver cancer tumors; melanoma tumors; lung cancer tumors; breast cancer tumors; colorectal cancer tumors; pancreatic cancer tumors; gastric cancer tumors; ovarian cancer, glioma and other malignant tumors tissue, the present invention also provides a pharmaceutical composition comprising the tumor targeting molecule; the present invention also provides a contrast agent for ultrasound imaging diagnosis and magnetic resonance imaging ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/14A61K49/22
Inventor 杨静雯
Owner 杨静雯
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