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Preparation method of DNA targeting nano medicine-carrying molecule for brain tumor

A DNA-targeting, nano-drug-loading technology, applied in anti-tumor drugs, pharmaceutical formulations, inactive components of polymer compounds, etc., can solve the problems of unseen patents, high penetration rate of brain malignant glioma, and insufficient drug treatment efficiency and other problems, to achieve the same size and structure, avoid damage, and overcome the effects of multi-drug resistance

Inactive Publication Date: 2015-05-27
SHANGHAI JIAO TONG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Brain malignant glioma seriously threatens human health due to its high penetration rate, recurrence rate and mortality rate. Traditional surgical treatment cannot completely remove tumor tissue, and the efficiency of drug treatment is less than 30%.
However, no relevant patents have been seen on DNA biomacromolecular materials combined with target molecules as drug carriers

Method used

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  • Preparation method of DNA targeting nano medicine-carrying molecule for brain tumor
  • Preparation method of DNA targeting nano medicine-carrying molecule for brain tumor
  • Preparation method of DNA targeting nano medicine-carrying molecule for brain tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] figure 1 Schematic diagram for the construction of the DNA-targeted nano-drug-carrying molecules, each take 2 μL of single-stranded DNA (50 μM) TSP-1, TSP-2, TSP-3, N 3 -TSP-4 was added to 42μL Tris-MgCl (Tris 10mM, MgCl 2 50mM, pH8) solution. The mixed solution was placed in a PCR instrument at a reaction temperature of 95° C., cooled rapidly to 4° C. after 10 minutes, and continued to react for 30 minutes. A single strand of DNA can self-assemble into a three-dimensional DNA tetrahedral configuration through complementary base pairing.

[0039] Take 180 μL of PBS solution (pH7.3), 40 μL of CuSO 4Aqueous solution (0.1 mM), 40 μL of TCEP aqueous solution (0.1 mM) and 40 μL of TBTA solution (10 μM, dissolved in DMSO) were prepared as a reaction solution. Take 100 μL of the prepared TDN solution (2 μM) and 200 μL of the target peptide solution (2 μM) whose amino acid sequence is RGERPPR, and add them to the above reaction solution, shake at 37°C for 5 hours at a const...

Embodiment 2

[0042] figure 1 Schematic diagram for the construction of the DNA-targeted nano drug-carrying molecules, each take 2 μL of single-stranded DNA (50 μM) TSP-1, TSP-2, Cy3-TSP-3, N 3 -TSP-4 was added to 42μL Tris-MgCl (Tris 10mM, MgCl 2 50mM, pH8) solution. The mixed solution was placed in a PCR instrument with a reaction temperature of 95° C., cooled to 4° C. after 10 minutes, and the reaction was continued for 30 minutes. The DNA single strand can self-assemble into a DNA tetrahedral three-dimensional configuration with a Cy3 fluorescent signal through complementary base pairing.

[0043] Take 180 μL of PBS solution (pH7.3), 40 μL of CuSO 4 Aqueous solution (0.1 mM), 40 μL of TCEP aqueous solution (0.1 mM) and 40 μL of TBTA solution (10 μM, dissolved in DMSO) were prepared as a reaction solution. Take 100 μL of the prepared TDN solution (2 μM) and 200 μL of the target peptide solution (2 μM) whose amino acid sequence is RGERPPR, and add them to the above reaction solution, ...

Embodiment 3

[0046] figure 1 Schematic diagram for the construction of the DNA-targeted nano-drug-carrying molecules, each take 2 μL of single-stranded DNA (50 μM) TSP-1, TSP-2, FAM-TSP-3, N 3 -TSP-4 was added to 42μL Tris-MgCl (Tris 10mM, MgCl 2 50mM, pH8) solution. The mixed solution was placed in a PCR instrument with a reaction temperature of 95° C., cooled to 4° C. after 10 minutes, and the reaction was continued for 30 minutes. DNA single strands can self-assemble into DNA tetrahedral three-dimensional configuration with FAM fluorescent signal through complementary base pairing.

[0047] Take 180 μL of PBS solution (pH7.3), 40 μL of CuSO 4 Aqueous solution (0.1 mM), 40 μL of TCEP aqueous solution (0.1 mM) and 40 μL of TBTA solution (10 μM, dissolved in DMSO) were prepared as a reaction solution. Take 100 μL of the prepared TDN solution (2 μM) and 200 μL of the target peptide solution (2 μM) whose amino acid sequence is RGERPPR, and add them to the above reaction solution, shake a...

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Abstract

The invention discloses a preparation method of a DNA (Deoxyribonucleic Acid) targeting nano medicine-carrying molecule for a brain tumor. The preparation method comprises the steps that a DNA single strand is added to a Tris-MgCl solution and mixed, and reacts to form a DNA tetrahedron solution; the DNA tetrahedron solution and a targeting peptide solution are added to a mixed solution of a PBS (Phosphoric acid solution), a CuSO4 solution, a TCEP (Tris-(2-carboxyethyl)-phosphine) solution and a TBTA (Tert-Butyl 2,2,2-trichloroacetimidate) solution, and subjected to thermostatic reaction; a targeting DNA tetrahedron solution is obtained; a tumor medicine is added to the targeting DNA tetrahedron solution; constant temperature oscillation, centrifugation, and supernatant removal are conducted; and an obtained sediment is the DNA targeting nano medicine-carrying molecule. According to the preparation method, peptide molecules having specificity in the tumor are modified on DNA tetrahedrons via a point-and-click reaction, so that construction of a targeting DNA nano-carrier is realized; and the targeting DNA medicine-carrying molecule has extremely high specific recognition function, low cytotoxicity and good structural stability.

Description

technical field [0001] The invention belongs to the field of life medicine, and relates to a preparation method of DNA-targeted nano drug-carrying molecules for brain tumors. Background technique [0002] Brain malignant glioma seriously threatens human health due to its high penetration rate, recurrence rate and mortality rate. Traditional surgical treatment cannot completely remove tumor tissue, and the efficiency of drug treatment is less than 30%. Once a tumor occurs, the blood-brain barrier in the human body will be destroyed, and the EPR effect, that is, the enhanced osmotic retention effect will allow more nanoparticles to pass through the blood-brain barrier. However, in the process from the peripheral tumor tissue to the internal tumor tissue, the EPR effect will gradually weaken, and the internal pore size will also decrease to 7-100nm. Therefore, in the study of brain tumors, higher requirements will be placed on the size and penetrability of materials. [0003]...

Claims

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Application Information

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IPC IPC(8): A61K47/42A61K31/704A61P35/00
Inventor 何丹农夏智伟王萍严一楠刘婷金彩虹
Owner SHANGHAI JIAO TONG UNIV
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