Application of medicine or product prepared by aptamer and used for treating multiple myeloma

A multiple myeloma and nucleic acid aptamer technology, applied in the fields of biomedicine and clinical medicine, can solve problems such as difficult treatment and poor treatment effect, and achieve high affinity and high specificity effects

Inactive Publication Date: 2015-02-25
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, traditional chemotherapy and hematopoietic stem cell transplantation are commonly used clinical treatments, but their therapeutic effect has been poor. In addition, the emergence of clinical multidrug resistance makes treatment more difficult, and the 5-year survival rate of patients is only 40%. Therefore, there is an urgent need to adopt new methods and means to treat MM patients and improve the survival period of patients.

Method used

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  • Application of medicine or product prepared by aptamer and used for treating multiple myeloma
  • Application of medicine or product prepared by aptamer and used for treating multiple myeloma
  • Application of medicine or product prepared by aptamer and used for treating multiple myeloma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] In order to clarify whether the nucleic acid aptamer TY04 has an effect on the proliferation of multiple myeloma cell lines, we used the CCK-8 method to detect the 4μM TY04 treatment of multiple myeloma cell lines (MM.1S, H929, KM3 , OPM2) and normal human immortalized B lymphocytes (the concentration of the tested cells was 1×10 5 Individuals / ml) The relative survival rate after 96h. After three independent repeated experiments, it was found that the relative survival rates (%) of normal human immortalized B lymphocytes and different multiple myeloma cells (MM.1S, H929, KM3, OPM2) treated with TY04 were: 144± 13.81, 47.72±5.7, 69.42±15.1, 58.23±0.65, 57.08±4.24. Compared with normal human immortalized B lymphocytes, TY04 can lead to a significantly lower survival rate of multiple myeloma cell lines, with a statistical difference (Pfigure 1 Generally speaking, if the IC50 value at the cell level is within 10 μM, it has anti-tumor development potential). Further use CC...

Embodiment 2

[0025] In order to clarify whether TY04 inhibits the proliferation of MM.1S has sequence specificity, we further tested multiple myeloma MM.1S cells treated with different concentrations of ssDNA library and TY04 (0, 0.5, 1.0, 2.0, 4.0 μM) The concentration of cells was 3 x 10 5 Individuals / ml or so) growth after 96h. After three independent repeated experiments, the data were statistically analyzed, and the results were as follows: image 3As shown, it shows that compared with TY04, the ssDNA library has no effect on the proliferation of multiple myeloma MM.1S cells, further indicating that the nucleic acid aptamer TY04 has sequence specificity for multiple myeloma MM.1S cells.

Embodiment 3

[0027] In order to clarify the binding of nucleic acid aptamer TY04 to multiple myeloma MM.1S cells, the nucleic acid aptamer TY04 with FITC fluorescent labeling was mixed with multiple myeloma Tumor MM.1S cells (the concentration of the test cells is 1×10 6 cells / ml) after incubation, the binding of TY04 to the surface of MM.1S cells was detected by flow cytometry. The result is as Figure 4 As shown, the fluorescent nucleic acid aptamer TY04 has significant binding to multiple myeloma MM.1S cells, and the binding strength increases with the increase of the concentration. In order to detect whether the binding of nucleic acid aptamer TY04 to multiple myeloma MM.1S cells is specific, the ssDNA library with FITC fluorescent labeling was mixed with multiple myeloma MM.1S cells at different concentrations (0, 25, 125 nM). After the cells were incubated, the binding of TY04 to the surface of MM.1S cells was detected by flow cytometry, and the binding of fluorescent aptamer TY04 ...

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Abstract

The invention provides the application of a medicine or a product prepared by aptamer and used for treating multiple myeloma. The aptamer adopts TY04, and belongs to single strand DNA containing 83 basic groups. The action principle of the aptamer is that the aptamer can present cell specificity and sequence specificity and be stably combined with protide substances on the surface of multiple myeloma cells, operate the expression of related protein of cyclin B, CDK1, ERK1 / 2 and gamma-tubulin by lowering the cell cycle, and retard the process of multiple myeloma cell cycle to G2 / M phase, thereby restraining the multiplication of the multiple myeloma cells. Presently, the aptamer for treating multiple myeloma is not reported at home and abroad. The discovery and application of the aptamer can provide a novel strategy for treating multiple myeloma.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and clinical medicine, and relates to the use of a nucleic acid molecule related to the treatment of multiple myeloma in the preparation of drugs or products for treating multiple myeloma. Background technique [0002] Nucleic acid aptamer (aptamer) is a short single-stranded DNA or RNA molecule with recognition function screened by an artificial screening technique that simulates the natural evolution process-exponential enrichment ligand system evolution (SELEX), and can form a specific DNA or RNA molecule by folding. The three-dimensional structure binds to the target molecule. It has the characteristics of high affinity, high specificity, non-immunogenicity, easy synthesis, transformation and labeling, good biochemical stability, reversible denaturation and renaturation, and can be amplified and cut by enzymes, especially some nucleic acids Aptamers can specifically bind to their target p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61P35/00
Inventor 刘静谭蔚泓叶茂宋健辉戴红娟
Owner CENT SOUTH UNIV
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