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Antisense oligonucleotide probe contrast agent marked by superparamagnetism iron oxide and production of the same

An antisense oligonucleotide and superparamagnetic technology, which is applied in the field of cancer gene targeting contrast agent and its preparation, can solve the problems of inability to display antisense activity, achieve clear anatomical structure, high signal-to-noise ratio, and achieve early Effect of Specific Diagnosis

Inactive Publication Date: 2010-01-20
CHONGQING MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, when the antisense oligonucleotide is less than 15bp, it cannot show antisense activity

Method used

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  • Antisense oligonucleotide probe contrast agent marked by superparamagnetism iron oxide and production of the same
  • Antisense oligonucleotide probe contrast agent marked by superparamagnetism iron oxide and production of the same
  • Antisense oligonucleotide probe contrast agent marked by superparamagnetism iron oxide and production of the same

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 preparation method

[0031] A non-limiting preparation method of the contrast agent of the present invention: comprising the following steps:

[0032] (1). Preparation of the carrier: one-step co-precipitation method is used to complete the synthesis of ferric oxide. The reaction principle is: Fe 2+ +2Fe 3+ +8OH - → Fe 3 o 4 +4H 2 O; adding dextran while synthesizing ferric oxide to obtain the ferric oxide carrier of the outsourcing dextran;

[0033] (2). The synthesis of the antisense oligonucleotide fragment of the c-erbB2 oncogene whose oligonucleotide sequence is CTCCATGGTGCTCAC. The antisense oligonucleotides in the examples were provided by Shanghai Sangon Co., Ltd.

[0034] (3). Synthesis of contrast agent: the specific steps are: the ferric iron tetroxide carrier coated with dextran obtained in the purification step (1): take 250 μL of the carrier after passing through the column; add 10 μL of 0.1M sodium periodate, The hydroxyl group of the...

Embodiment 2

[0036] Embodiment 2 biological and physical properties detection:

[0037] See attached figure 2 , the order of the 5 spotting wells in the figure is from left to right: the 1st and 2nd wells are antisense oligonucleotide standard products 1ug and 3ug, the 3rd well is the contrast agent of the present invention, and the 4th and 5th wells are carriers Mixed with antisense oligonucleotides. It can be seen from the figure that the contrast agent of the present invention has a high degree of purification and stable properties.

[0038] See attached image 3 , where Figure A is the spectrum of the antisense oligonucleotide standard, and Figure B is the spectrum of the vector, and their peak shapes are good without interference from other peaks. Figure C is the mixture of carrier and antisense oligonucleotides. The two are completely separated, and the peak shape is good without tailing. The recovery rate is 100%, which proves that the gel column used in this experiment can be u...

Embodiment 3

[0040] Embodiment 3 Magnetic property detection:

[0041] See attached Figure 5 Through oscillating sample magnetometer and magnetic resonance detection, the saturation magnetization of the sample is 69.32875emu / g Fe, the specific saturation magnetization is 68.41215emu / g, the specific residual magnetization is 30.4753emu / g, the residual magnetization is 19.25782Gs, and the relaxation The rate is 0.154×10 6 mol -1 sec -1 . The magnetization curve is a single curve passing through the origin, indicating that the contrast agent of the present invention has superparamagnetism.

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Abstract

The invention discloses a trans-oligonucleotide probe contrast-medium to mark superparamagnetism iron oxide, which consists of carrier constituted by gluglucosan enveloped by ferroferric oxide nanometer particle with superparamagnetism and trans-oligonucleotide segment of c-erbB2 cancer gene, wherein the trans-oligonucleotide connects the gluglucosan in the carrier at covalent bond. The invention also relates to a making method of the contrast-medium. The invention uses trans-gene technique of molecular biological domain into imaging diagnosis to combine the superiority and specificity of two advanced techniques, which increases the ratio of target / non-target of probe to the maximum degree to scan the size, position and anatomical relationship of adjacent structures of tumour through magnetic resonance, in order to build new imaging method of early-stage tumor diagnosis of specificity on the gene level. The invention can be applied to do early-stage imaging diagnosis for malignant tumour, epoophoron cancer, uterine neck cancer, esophagus cancer and cervical scale cancer.

Description

technical field [0001] The invention relates to the technical field of medical detection reagents, in particular to an oncogene targeting contrast agent capable of specifically imaging tumors in magnetic resonance imaging and a preparation method thereof. Background technique [0002] Malignant tumors are common and frequently-occurring diseases that threaten human health, and are increasing year by year. Early and specific diagnosis of malignant tumors is the key to improving the survival rate and quality of life of patients. Traditional imaging diagnosis methods are mainly aimed at imaging after the tumor has formed a solid mass, but at this time the patient is already in the middle and late clinical stage, the treatment effect is not good, and the prognosis is poor; in addition, the contrast agent gadolinium ion currently used for magnetic resonance imaging It is an extracellular non-specific contrast agent, which stays in the human body for a short time, and cannot be us...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K49/06A61K49/08
Inventor 文明李少林李必波柏玮彭志平黎孝平
Owner CHONGQING MEDICAL UNIVERSITY
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