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Preparation and applications of pH and ATP response-type nano carrier

A nano-carrier and carrier technology, which is applied in the field of biomedicine, can solve the problems of lack of tissue-specific toxicity, and achieve the effects of efficient and targeted inhibition of tumor proliferation, increased biocompatibility, and efficient and rapid release

Active Publication Date: 2019-06-04
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The effectiveness of these delivery systems has been proven, however they are mostly exogenous, lack tissue specificity and are potentially toxic

Method used

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  • Preparation and applications of pH and ATP response-type nano carrier
  • Preparation and applications of pH and ATP response-type nano carrier
  • Preparation and applications of pH and ATP response-type nano carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Preparation of pH and ATP-responsive nano drug delivery system:

[0047] Preparation of the main reaction solution:

[0048] Annealing buffer (200mM KCl, 4mM MgCl 2 , 28mM Tris-HCl): Accurately weigh 0.0190g MgCl 2 , 0.7456g KCl, 0.1695g Tris, add an appropriate amount of water to dissolve, adjust the pH to 8.0 with hydrochloric acid, and dilute to 50mL.

[0049] 1. Nucleic acid sequence annealing

[0050] ATP aptamer-i-motif and cDNA single-stranded DNA dry powder were added to nuclease-free water, respectively, and dissolved into nucleic acid solutions with a concentration of 100mM.

[0051] Add ATP aptamer-i-motif solution, cDNA solution, and annealing solution into the PCR tube at a ratio of 1:1:1 and shake and mix evenly. The mixed liquid is annealed in the PCR amplification instrument (heating at 95°C for 10mins, then slowly lowering to room temperature) to form a stem-loop structure of an ATP aptamer double helix structure (ATP-aptamer-cDNA, hereina...

Embodiment 2

[0057] Example 2 Investigation of the Related Properties of pH and ATP Responsive Nanocarriers

[0058] 1. Investigation of the responsive release of the nanosystem Aaim@DNM

[0059] ATP response: Dilute Aaim@DNM to 2.5ml with PBS buffer solution with a pH of 7.4, measure its fluorescence value, then add a certain amount of 400mM ATP solution one by one, the fluorescence measurement parameters are the same as in Example 1, and measure the change of its fluorescence intensity. The result is as Figure 3-a shown. Similarly, the detection result of the fluorescence intensity change at pH=5.6 is as follows Figure 3-b shown.

[0060] Depend on Figure 3-a and Figure 3-b It can be seen that with the increase of ATP, the greater the change of fluorescence intensity, the more DNM released by Aaim@DNM.

[0061] pH response: Dilute Aaim@DNM to 2.5 ml with PBS buffer at pH 3.8, 5.6, 6.8, and 7.4, respectively, and measure its fluorescence value. Fluorescent detection results su...

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Abstract

The invention provides preparation and applications of a pH and ATP response-type nano carrier. The pH and ATP response-type nano carrier comprises a nucleic acid nano carrier and an anti-tumor drug immobilized on the nucleic acid nano carrier; the anti-tumor drug is anthracycline antibiotics; and the nucleic acid nano carrier comprises an ATP aptamer double helix structure and an i-motif stem loop structure. The pH and ATP response-type nano carrier provided by the invention utilizes the characteristics of tumor cell high-concentration ATP to promote dissociation of aptamers and complementarysequences thereof; and utilizes the acidic environment of tumor cells to promote the i-motif conformational change and to realize the efficient and rapid release of drugs in cells.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the preparation and application of a pH- and ATP-responsive nano-carrier. Background technique [0002] Chemotherapy and radiotherapy are the main means of cancer treatment. Traditional small-molecule chemical drugs have good therapeutic effects, but have disadvantages such as poor water solubility, low bioavailability, and strong toxicity, and require large doses of drug administration, which brings great toxic and side effects to patients (Chari R V J, Miller M L, Widdison W C. ChemInform Abstract: Antibody—Drug Conjugates: An Emerging Concept in Cancer Therapy [J]. Cheminform, 2014, 45(22).). Taking the powerful anti-tumor drug daunomycin as an example, it mainly forms a stable complex by intercalating into the double strand of DNA, affecting the structure and function of DNA, and preventing DNA replication and RNA synthesis. However, due to its lack of tumor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/51A61K47/26A61K31/704A61P35/00
Inventor 赵平林慧超
Owner GUANGDONG PHARMA UNIV
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