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A tumor-targeted gadolinium-based magnetic resonance imaging contrast agent and its preparation method

A contrast agent and gadolinium-based technology, applied in the field of contrast agents, can solve problems affecting clinical application, short residence time, increased injection dose, etc., and achieve good biocompatibility, good biocompatibility, and reduced dosage Effect

Active Publication Date: 2021-05-04
INST OF GEOLOGY & GEOPHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this type of contrast agent has a short retention time in the body. In order to achieve better imaging results, clinicians have to increase the injection dose, which increases the risk of side effects of free Gd ions in the body causing fibrosis to the kidneys, seriously affecting the kidneys. Clinical application

Method used

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  • A tumor-targeted gadolinium-based magnetic resonance imaging contrast agent and its preparation method
  • A tumor-targeted gadolinium-based magnetic resonance imaging contrast agent and its preparation method
  • A tumor-targeted gadolinium-based magnetic resonance imaging contrast agent and its preparation method

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Experimental program
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Embodiment 1

[0037] The preparation of embodiment 1, HFn-Gd

[0038] Using the full-length cDNA of human H subunit ferritin (HFn) as the amplification template, the HFn gene was obtained by polymerase chain reaction, and constructed on the pET11b plasmid, and then the constructed complete plasmid was transformed into Escherichia coli engineering bacteria BL21 , by culturing bacteria, inducing expression, disrupting bacteria, and separating and purifying to obtain recombinant HFn protein, the amino acid sequence of which is shown in SEQ ID NO:1.

[0039] Prepare DTPA and Gd ion solution with the same molar mass, stir and mix well to form the gadolinium chelate Gd-DTPA, use HFn and gadolinium chelate to condense the gadolinium chelate through the amide condensation reaction of the carboxyl group of the chelating agent and the amino group of the protein Connected to the surface of HFn, in the presence of condensing agents EDC and NHS, mix the gadolinium chelate with HFn, so that DTPA and HFn ...

Embodiment 2

[0040] The inspection of embodiment 2, HFn-Gd stability

[0041] In order to test the stability of HFn-Gd and confirm that there is no free Gd ion in the solution and serum, add the sample and arsenazoШ solution together, the free metal ion will combine with the solution and form an absorption peak at 650 nanometers, by Judging whether the arsenazoIII solution has an absorption peak at 650 nm indicates whether there is free Gd in the sample. figure 2 a shows the arsenazoIII solution (black line), arsenazoIII added commercial Gd-DTPA (blue line), freshly prepared HFn-Gd (green line) and HFn-Gd (red line) stored for two months The UV / Vis spectral absorption figure of sample, all samples have no absorption peak at 650 nanometers, have shown the stability of HFn-Gd preparation. figure 2 b shows the arsenazoIII solution (black line), arsenazoIII added fetal bovine serum (FBS, blue line), pre-incubated with FBS and commercial Gd-DTPA (red line) and pre-incubated with FBS and HFn-...

Embodiment 3

[0042] The longitudinal relaxation rate (r of embodiment 3, HFn-Gd and commercial Gd-DTPA 1 ) determination

[0043] The HFn-Gd material was first diluted to a final Gd concentration of 0-1 mM, and then assayed in an MRI system (Bruker, BioSpec70 / 20USR). Material T 1 The value is determined using a multi-slice-multi-echo sequence for T 1 Weighted imaging, the specific parameters are field of view (FOV) = 4cm × 4cm, matrix = 256 × 256, repetitiontime (TR) = 150ms, and TE = 1.21 ms (T 1 ). T of different concentrations of HFn-Gd 1 The reciprocal of the value and its concentration value are linearly fitted to obtain the r of the material 1 value. image 3 a is the measured r of different batches of HFn-Gd materials under a magnetic resonance instrument with a field strength of 7T 1 value, an average of 4.78mM -1 the s -1 , image 3 b for its T 1 Magnetic resonance imaging image, the greater the concentration, the lower the signal value and the brighter the image. ima...

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Abstract

The invention discloses a preparation method of a gadolinium-based magnetic resonance imaging contrast agent with tumor targeting and its biomedical application. The contrast agent can specifically bind to transferrin receptor 1 highly expressed on the surface of tumor cells, is composed of genetically engineered recombinant protein and gadolinium chelate, and has a higher activity than the commercial contrast agent Magnevist (Gd-DTPA). longitudinal relaxation rate. The present invention provides a comparison of the imaging effects of animal tumor models with Magnevist, indicating that the contrast agent in the present invention can achieve tumor-specific recognition and longer-term magnetic resonance imaging at a lower injection dose without repeated injections. The contrast agent in the present invention can be administered intravenously, and is applied to the early diagnosis of cancer, the further identification of tumor metastasis and the diagnosis of suspicious recurrence after operation.

Description

technical field [0001] The invention relates to the technical field of contrast agents, in particular, the invention discloses a tumor-targeting gadolinium-based magnetic resonance imaging contrast agent and a preparation method thereof. Background technique [0002] According to the World Health Organization, cancer is the second leading cause of death in the world, causing 8.8 million deaths in 2015. Malignant tumors seriously threaten human health and life. According to data from the World Cancer Foundation, in 2018, there were an estimated 18 million cancer cases worldwide, of which 9.5 million were men and 8.5 million were women. Cancer prevention has become one of the most significant public health challenges in the 21st century. In China, the "National Cancer Report 2019" released by the National Cancer Registry Center shows that an average of more than 10,000 people are diagnosed with cancer every day, and 7.5 people are diagnosed with cancer every minute. The situat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K49/14
CPCA61K49/085A61K49/14
Inventor 蔡垚潘永信张同伟曹长乾田兰香
Owner INST OF GEOLOGY & GEOPHYSICS CHINESE ACAD OF SCI
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