Radionuclide marked microRNA-155 targeting probe and application thereof in tumor imaging

A radionuclide and probe technology, applied in the biological field, can solve the problems that depend on the effective selection of pathological tissues, are not suitable for screening and prevention, and cannot meet the needs of early diagnosis

Inactive Publication Date: 2015-05-20
PEKING UNIV FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These detection methods have the following problems: invasive, require puncture biopsy tissue or surgery to remove the lesion, and the operation process is relatively cumbersome, the result depends on the effective s

Method used

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  • Radionuclide marked microRNA-155 targeting probe and application thereof in tumor imaging
  • Radionuclide marked microRNA-155 targeting probe and application thereof in tumor imaging
  • Radionuclide marked microRNA-155 targeting probe and application thereof in tumor imaging

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Experimental program
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Effect test

Embodiment 1

[0070] Embodiment 1, miR-155 targeting probe 99m Preparation of Tc-AAM-155

[0071] miR-155 targeting probe 99mFor the specific synthesis and reaction scheme of Tc-AAM-155, see figure 1 , is to chemically modify the antisense oligonucleotide sequence of miR-155, these chemical modifications include the phosphatidyl (PS) modification of the phosphate backbone on each of the 6 bases at both ends of the probe and the 3 bases at each of the two ends of the probe A base was modified with 2'-methoxy (2'-OMe), and it was radionuclides by the bifunctional chelating agent S-acetyl-NHS-MAG3 (N-hydroxysuccinimidyl derivative of S-Acetyl mercaptoacetyltriglycine) 99m Tc mark.

[0072] targeted probe 99m The preparation method of Tc-AAM-155 is as follows:

[0073] 1. Acquisition of miR-155 target nucleic acid after chemical modification

[0074] 1. Determination of miR-155 target nucleic acid sequence

[0075] Determine the antisense oligonucleotide sequence of miR-155 as the experi...

Embodiment 2

[0123] Embodiment 2, miR-155 targeting probe 99m Functional verification of Tc-AAM-155

[0124] 1. Evaluation of serum stability

[0125] The miR-155 targeting probe prepared in Example 1 99m Tc-AAM-155 was placed in human fresh serum for 12 hours at room temperature, its radiochemical purity was determined by HPLC and its nucleic acid stability was determined by gel electrophoresis.

[0126] 1. Nucleic acid serum stability

[0127] Determination of probe stability within 12h in fresh human serum (0h, 2h, 4h, 6h, 8h, 10h, 12h)

[0128] (1) Take 80 μl-120 μl of human fresh serum and divide it into 8 parts;

[0129] (2) Will 99m Tc-AAM-155 (chemically modified AAM-155, the total amount is 4nmol) was dissolved in 45μl buffer, dissolved in serum according to the volume ratio of 1:2 or 1:3, each sample was about 440pmol, and the sample was added in reverse order. Add the sample at 12h first, and add the sample at 0h before running the glue. Mix well and let stand at room tem...

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Abstract

The invention discloses a radionuclide marked microRNA-155 targeting probe for tumor imaging. According to the microRNA-155 targeting probe, the terminal 5' of a microRNA-155 targeting nucleic acid is connected with an amino acid by use of a hexyl; the nucleotidesequence of the microRNA-155 targeting nucleic acid is sequence 2 in the sequence table; the first to sixth nucleotides and the 18th to 23th nucleotides from the terminal 5' of the sequence 2 are all modified by sulfo groups, and the first to third nucleotides and the 21th to 23th nucleotides from the terminal 5' are all modified by 2'-methoxyls. The 99mTc-AAM-155 targeting probe can be effectively applied to vital tumor imaging, and also is capable of tracing the distribution situation in vivo of miR-155 at the vital level.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a radionuclide-labeled microRNA-155 targeting probe and its application in tumor imaging. Background technique [0002] Malignant tumors are a class of diseases that seriously endanger human health. According to statistics, there are about 1.6 million new cancer patients in my country every year, and about 1.3 million people die from cancer every year. Early diagnosis of tumors can enable patients to receive timely treatment, increase the cure rate, improve prognosis, and reduce mortality. Therefore, the "three early" principles of early detection, early diagnosis, and early treatment have been widely valued in tumor diagnosis and treatment, and have always been important in tumor research. hot topics. Among the many diagnostic methods, the radionuclide tracer technology of nuclear medicine is unique due to its high sensitivity, non-invasiveness, dynamic monitoring of living body a...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10A61K51/06A61K103/10
Inventor 康磊王荣福徐小洁闫平张春丽霍焱
Owner PEKING UNIV FIRST HOSPITAL
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