A kind of APD hybrid nano system and its construction method and application

A construction method and nanotechnology, applied in the fields of nanotechnology, nanotechnology, nanomedicine, etc., can solve the problems of easy aggregation of gold rods, complicated operation, tedious operation, etc., and achieve the advantages of cell endocytosis, simple preparation method, and cell entry efficiency. high effect

Active Publication Date: 2021-01-05
SICHUAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] 2) The right-handed DNA tetrahedron will be degraded by deoxyribonuclease, and its stability is poor, which is not conducive to its long-term stable function
[0009] 3) The combination reaction of thiolated TDNs and gold nanorods has problems such as cumbersome operation, time-consuming, and easy aggregation of gold nanorods; the modification process of gold nanorods is complicated and cumbersome, and requires experimenters to have certain experience and skills, otherwise, it is easy to cause The modification of gold nanorods failed; the modification process of gold nanorods took a long time, ranging from a few hours to a day or two, which took a lot of precious time of the experimenters; at the same time, in the experiments or treatments that depended on DNA-modified gold nanorods, It takes a lot of time to prepare DNA-modified gold nanorods, with poor timeliness and low work efficiency

Method used

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  • A kind of APD hybrid nano system and its construction method and application
  • A kind of APD hybrid nano system and its construction method and application
  • A kind of APD hybrid nano system and its construction method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] In this embodiment, left-handed DNA is also expressed as L-DNA, and "L-" is used to indicate left-handed.

[0060] In this example, the APD hybrid nanosystem is prepared through the following steps.

[0061] (1) Mix 3.7 mL of 0.1 mol / L CTAB and 150 μL of 0.01 mol / L HAuCl 4 After mixing, add 0.85mL deionized water, then add 0.3mL 0.01 mol / L ice NaBH in ice bath 4 Aqueous solution, 1200 r / min rapid stirring for 2 minutes, standing for 2 hours will immediately generate CTAB-coated gold seeds. Add 1.4 mL of the prepared gold seed solution into the gold nanorod growth solution, stir rapidly at 450 rpm for 2 minutes, let it stand for 12 hours, then centrifuge at 10,000 rpm for 10 minutes, and resuspend with 0.01 mol / L CTAB solution Gold nanorods, stored at room temperature for future use. The growth liquid is: 300mLCTAB, the concentration of the substance is 0.1 mol / liter; 16mLHAuCl 4 , the concentration of the substance is 0.01 mol / L; 3mLAgNO 3 , the concentration of th...

Embodiment 2

[0071] This example mainly illustrates the anti-nuclease degradation ability of the APD nanosystem.

[0072] Using the same method to prepare left-handed DNA tetrahedron and right-handed DNA tetrahedron, the preparation method is the same as step (2) of Example 1. Right-handed DNA (D-DNA) was used to prepare right-handed DNA tetrahedrons (D-TDNs), and the base sequence of D-DNA was the same as that of L-DNA. The preparation method of APD is the same as that of Example 1, and will not be repeated in this example. D-TDNs, L-TDNs and APD were mixed with deoxyribonucleotide enzymes, incubated at 37°C for 8h, and EDTA was added to terminate the enzymatic degradation reaction. Subsequently, heparin sodium was added to APD and enzyme to destroy the electrostatic interaction and release D-TDNs. The remaining samples were not treated with sodium heparin.

[0073] Using polyacrylamide gel electrophoresis test to observe the movement of the sample, the results are as follows Figure ...

Embodiment 3

[0075] This example is an example of preparation of a targeted APD nanosystem.

[0076] Synthesis of folic acid-mercapto-containing polylysine conjugates: Feed folic acid, mercapto-containing polylysine, EDC and NHS at a molar ratio of 1:1:1.5:1.5, dissolve in 20ml of dimethyl sulfoxide, and stir magnetically at room temperature 24 hours. Then the reaction solution was transferred to a dialysis bag with a cut-off molecular weight of 6000 Da, immersed in ultrapure water for dialysis for 24 hours. When the dialysis time expired, the moisture in the sample was removed by freeze-drying to obtain a folic acid-polylysine conjugate.

[0077] Follow the steps below to prepare the nanosystem:

[0078](1) Prepare gold nanorods of specific size by seed crystal growth method. Add 3.7 mL of 0.1 mol / L CTAB and 150 μL of 0.01 mol / L HAuCl 4 After mixing, add 0.85mL deionized water, then add 0.3mL 0.01 mol / L ice NaBH in ice bath 4 Aqueous solution, stirred rapidly at 1200rpm for 2min, and...

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Abstract

The invention discloses an APD hybrid nano system and its construction method and application. The hybrid system is obtained by mixing and incubating nano-gold rods modified with thiol-containing cationic polymers and left-handed DNA tetrahedrons. The invention uses unmodified DNA, avoids the use of expensive thiol DNA, and is more conducive to reducing the cost of nanoscale drug carriers. The inventors have discovered and proved through experiments that the use of left-handed DNA can resist ribozyme degradation, and at the same time, after the polymer and DNA are compounded, the DNA can be protected, and the dual functions improve the stability of the system. The APD hybrid nanosystem also has the ability to enter cells efficiently. It can be used to load drugs, especially chemotherapy drugs, and has application prospects in the field of combined therapy of tumor hyperthermia and chemotherapy.

Description

technical field [0001] The invention belongs to the field of nanoscale drug carriers, relates to a hybrid nano system, and also relates to a construction method and application of the hybrid nano system. Background technique [0002] Professor Russell P. Goodman of the University of Oxford in the United Kingdom invented the "one-step method" to synthesize DNA tetrahedrons (TDNs), which is easy to operate and has a high yield. DNA tetrahedron has the advantages of good biocompatibility, strong loading capacity, designable structure, and uniform size. DNA tetrahedron can load a variety of chemotherapy drugs, DNA, RNA, and oligonucleotides, etc., and can be used in biosensors, biomolecules, etc. Detection, drug carrier and other biomedical fields. [0003] Gold nanorods have anisotropic shape and exhibit both transverse and longitudinal plasmon resonance (SPR) effects. By changing the aspect ratio of gold nanorods, the longitudinal SPR peak (LSPR) can be precisely tuned in th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/00A61K47/18A61K41/00A61K31/704A61P35/00B82Y5/00
CPCA61K9/0092A61K31/704A61K41/0052A61K47/186A61P35/00B82Y5/00A61K2300/00
Inventor 李莉燕建芹陈军何斌
Owner SICHUAN UNIV
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