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PH-induced drug sustained-release deoxyribonucleic acid (DNA) nanostructure as well as preparation method and application thereof

A nanostructure and DNA molecule technology, applied in DNA preparation, biochemical equipment and methods, recombinant DNA technology, etc., can solve problems such as influence and side effects, and achieve the effect of high pH response sensitivity, rapid response, and increased drug loading.

Inactive Publication Date: 2019-04-26
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, one of the disadvantages of chemotherapy is that while anticancer drugs attack tumor cells, they also affect non-pathological normal cells and cause serious side effects

Method used

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  • PH-induced drug sustained-release deoxyribonucleic acid (DNA) nanostructure as well as preparation method and application thereof
  • PH-induced drug sustained-release deoxyribonucleic acid (DNA) nanostructure as well as preparation method and application thereof
  • PH-induced drug sustained-release deoxyribonucleic acid (DNA) nanostructure as well as preparation method and application thereof

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preparation example Construction

[0035] In a first typical embodiment of the present disclosure, a method for preparing a pH-induced drug sustained-release DNA nanostructure is provided, the method comprising the following steps:

[0036] Synthesis of long single-stranded DNA by rolling circle amplification reaction;

[0037] Complementary base pairing combination with another G- and C-rich DNA single strand (loading strand or loading strand) to obtain DNA molecular aggregates with alternating single and double strands, which is a pH-induced slow-release drug DNA nanostructure;

[0038] Wherein, the double-stranded part of the DNA molecular aggregate contains a site capable of loading drugs;

[0039] When the pH is lowered, the rolling circle amplification product is triggered to fold to form a triple helix configuration, the double strand of the DNA molecular aggregate unwinds, and the G and C-rich DNA single strand (loading strand or loading strand) is released, and the intercalated DNA is released. drug. ...

Embodiment 1

[0075] Preparation method of a DNA nanostructure for pH-induced sustained drug release

[0076] Experimental principle such as figure 1 As shown, long single-stranded DNA structures are formed by rolling circle amplification reactions. The template strand is a DNA single strand with a length of 81 bases modified with a phosphate group at the 5' end. There are 14 and 15 bases at the 5' end and 3' end respectively, which are complementary to the primer strand and annealed together with the primer strand. After gradually cooling down to 25°C, the template chain and the primer chain are hybridized and circularized. Under the action of T4 DNA ligase and ATP, the phosphate group at the 5' end of the template chain and the hydroxyl group at the 3' end form a phosphodiester bond, thus forming a rolling circle. Amplified circular template. After adding phi29 DNA polymerase and dNTPs, the rolling circle amplification reaction is initiated to form long single-stranded DNA. The loading...

Embodiment 2

[0078] A pH-induced drug sustained-release DNA nanostructure, the method for realizing the slow-release of anticancer drug doxorubicin mainly includes the following steps:

[0079]1. Prepare a circular template: first, the primer strand (primer strand) and the padlock probe (template strand) with a phosphate group modified at the 5' end are mixed in 1×T4 DNA ligase buffer (40mM Tris-HCl, 10mM MgCl 2 , 10mM DTT, 0.5mMATP, pH 7.8) annealed together. The annealing procedure was to heat to 95°C for 5 minutes, then cool to 25°C at 0.1°C / s and continue to stabilize for 3 hours. Then add T4 DNA ligase to the hybridized DNA solution, react at 16° C. for 12 hours, and then heat at 65° C. for 10 minutes to inactivate T4 DNA ligase.

[0080] 2. Rolling circle amplification to synthesize long single-stranded DNA: add 1×DNA polymerase buffer (33mM Tris-acetate, 10mMMg(CH 3 COO) 2 , 66mM CH 3 COOK, 1% Tween 20, 1mM DTT, pH7.9), add the prepared circular template solution, phi29DNA polym...

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Abstract

The disclosure relates to a pH-induced drug sustained-release deoxyribonucleic acid (DNA) nanostructure as well as a preparation method and application thereof. Long single-stranded DNA is synthesizedby using a rolling circle amplification technology, and is complemented and hybridized with G and C-rich DNA single strand (loading strand) so as to obtain DNA molecular aggregates with alternating single and double strands; the DNA molecular aggregates can carry a large amount of Dox. When the pH of the system decreases, the rolling circle amplification product is folded to form a triple helix configuration, the double strands of the DNA molecule aggregates are melted, the G and C-rich DNA single strand is released, and the inserted Dox is also released, so that the drug release is completed. The reversible intercalation and release of the Dox can be achieved by adjusting the pH. The method utilizes the DNA nanostructure with good biocompatibility as a drug carrier to increase the drug loading amount, and has the advantages of being low in cost, simple to operate, high in sensitivity to pH response, rapid in response, and the like.

Description

technical field [0001] The disclosure relates to a pH-induced drug slow-release DNA nanostructure and its preparation method and application. Background technique [0002] The information disclosed in this Background section is only intended to increase the understanding of the general background of the disclosure, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes prior art that is already known to those skilled in the art. [0003] Chemotherapy is the main way to treat cancer at present, but chemotherapy drugs will spread throughout most of the tissues and organs of the whole body along with the blood circulation. Therefore, one of the disadvantages of chemotherapy is that while anticancer drugs attack tumor cells, they also affect non-pathological normal cells and cause serious side effects. For example, doxorubicin (Dox), an anthracycline widely used in cancer chemotherapy, can induce cardiomyopathy leading...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/54A61K31/704A61P35/00B82Y5/00C12N15/10
CPCC12Q1/6844A61K31/704A61K47/549A61P35/00B82Y5/00C12Q2531/125C12Q2525/307
Inventor 毕赛李园芳岳淑珍潘思羽
Owner QINGDAO UNIV
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