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Construction and application of nucleic acid aptamer that can bind to CD123 and its targeted drug delivery system

A nucleic acid aptamer and drug-carrying system technology, applied in the medical field

Active Publication Date: 2020-05-19
XIAN CHILDRENS HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no reports of CD123 nucleic acid aptamers at home and abroad

Method used

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  • Construction and application of nucleic acid aptamer that can bind to CD123 and its targeted drug delivery system
  • Construction and application of nucleic acid aptamer that can bind to CD123 and its targeted drug delivery system
  • Construction and application of nucleic acid aptamer that can bind to CD123 and its targeted drug delivery system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0023] 1. Screening: Use the SELEX method to screen nucleic acid aptamers that can bind to CD123

[0024] a) Main materials and reagents

[0025] cell line

[0026] Human acute myeloid leukemia MOLM-13, human blood leukemia stem cell line TF-1, human B-cell precursor leukemia RCH-ACV, mouse lymphoma cell line EL4.

[0027] bacteria

[0028] Competent cells Trans1-T1: purchased from Beijing Quanshijin Biotechnology Co., Ltd., long-term storage at -80°C, try to avoid repeated freezing and thawing.

[0029] b) Construction of DNA library and primers in vitro:

[0030] Design a DNA library containing random sequences in vitro by software such as Primer 5, so that the number of clones reaches 10 15 -10 18 One, to ensure the polymorphism of the nucleic acid aptamer; synthesize primers paired with the library, and modify biotin (Biotin) and fluorescein isothiocyanate (fluorescein isothiocyanate, FITC) at the 5' end of the primers respectively, for SELEX screening Prepare.

[0...

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PUM

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Abstract

The invention relates to the medical field and particularly relates to an aptamer capable of being bound with CD123, construction of a targeting drug delivery system of the aptamer and application of the aptamer. The aptamer is named as ZW25 and has a sequence represented by SEQ ID NO.1: 5'-TGCGTGTGTAGTGTGTCTGGGCTACATCGATGAGCTGCCTAGGGTCCCTCTTAGGGATTTGGGCGG-3'. The aptamer can be rapidly synthesized in quantity in vitro and has the advantages that a preparation method is simple, the modification is easy, carrier drugs can be easily carried, the stability is superior to that of an antibody, the storage is facilitated, tumor tissues are easily penetrated by the aptamer, the immunogenicity is low, and the like.

Description

technical field [0001] The invention relates to the field of medicine, in particular to the construction and application of a nucleic acid aptamer capable of binding to CD123 and its targeted drug delivery system. Background technique [0002] Acute leukemia in children is the most common type of malignant tumors in children and the leading cause of death in children due to diseases. It can be divided into acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). At present, only about 45% of children with AML have long-term disease-free survival, and the prognosis is worse than that of ALL. Relapse after chemotherapy and hematopoietic stem cell transplantation is a difficult point in the treatment of AML by modern medicine. Studies suggest that leukemia stem cells (LSCs) are an important cause of relapse. LSCs can self-replicate and renew, and conventional chemotherapy drugs are ineffective against them. Therefore, residual LSCs in patients with AML after treat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115A61K47/54A61K31/704A61P35/02
CPCA61K31/704C12N15/115C12N2310/16
Inventor 张李钰李安茂张艳敏武海滨杨颖李俏王猛
Owner XIAN CHILDRENS HOSPITAL
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