Nanocomposite carrier for transfection, nucleic acid nanomedicine and preparation method and application thereof

A nanocomposite, nucleic acid nanotechnology, applied in the field of biomedicine, can solve the problems of limited application of siRNA delivery siRNA drugs, unstable lipid nanocarriers, etc., to improve transfection performance, avoid toxic side effects, and high therapeutic efficiency. Effect

Active Publication Date: 2022-07-19
INST OF CHEM CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problem that siRNA carriers in the prior art are difficult to deliver siRNA into organs other than the liver, resulting in limited application of siRNA drugs, the purpose of the present invention is to provide a nanocomposite carrier for transfection, nucleic acid nanomedicine and In its application, the nanocomposite carrier is used to load siRNA, protect siRNA and at the same time assist siRNA to cross the lung tissue barrier, so as to achieve efficient transfection (especially lung tissue transfection), and solve the problem of most lipid nanocarriers in lung tissue mucus. Instability issues in layers

Method used

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  • Nanocomposite carrier for transfection, nucleic acid nanomedicine and preparation method and application thereof
  • Nanocomposite carrier for transfection, nucleic acid nanomedicine and preparation method and application thereof
  • Nanocomposite carrier for transfection, nucleic acid nanomedicine and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: siRNA / Tetrapiperazine Epoxy C 60 Preparation of Derivatives / Polythiophene Polymer Nucleic Acid Nanomedicines

[0045] (a) Take 1 mL of 1 mM tetrapiperazine epoxy C 60 Derivatives (TPFE) aqueous solutions were added with different moles of 1 mL of polythiophene (PT), and the thiophene monomer concentrations were 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, and 8 mM PT aqueous solution, and mixed well by a shaker. , after standing for 3 minutes, use a 0.22 μm water-soluble filter membrane to filter, and finally obtain the molar ratio of TPFE and PT monomer of 1:1, 1:2, 1:3, 1:4, 1:5, 1: 6, 1:7, 1:8 nanocomposite carrier aqueous solution. The zeta potential of each nanocomposite carrier was measured by taking 1 mL of each solution using the Zetasizer Nano ZSP instrument, see figure 1 . From the measurement results of zeta potential, it can be seen that with the increase of PT molar amount, the zeta of the nanocomposite carrier gradually changes from positive t...

Embodiment 2

[0048] Example 2: Detection of hydrated particle size of nanomedicine and individual components

[0049] Take 1 mL of 0.01 mM TPFE aqueous solution, 0.02 mM PT aqueous solution of polythiophene monomer molar concentration, 0.01 mM TPFE-containing nanocomposite carrier prepared in Example 1 of the present invention, and 0.01 mM TPFE prepared in Example 1 of the present invention. The hydrated particle size of each material in ultrapure water was determined by dynamic light scattering (DLS), see figure 2 . Due to the electrostatic interaction and π-π interaction between the ternary components, the nucleic acid nanomedicine will self-assemble to form nanoparticles in water. The polydispersity of TPFE alone in water is relatively high, and it fails to form stable and uniform nanoclusters. The average hydrated particle size of PT alone in water is 362.8 nm. The hydrated particle size is 119.6 nm, and the average hydrated particle size of the nucleic acid nanomedicine composed of...

Embodiment 3

[0050] Example 3: Biosafety of nanocomposite carriers detected by CCK-8 method

[0051] The cytotoxicity of TPFE, PT and nanocomposite carrier (T&P) complexed with TPFE:PT monomer molar ratio of 1:2 on mouse melanoma cells (B16F10) was detected by CCK-8 method.

[0052] 96-well plate, 1x10 per well 4 When the confluence reaches 40% to 50%, add 200 μL of 25 μM, 50 μM, 75 μM, 100 μM, 125 μM, 150 μM TPFE and 50 μM, 100 μM, 150 μM, 200 μM, 250 μM, 300 μM PT and PT containing 25μM, 50μM, 75μM, 100μM, 125μM, 150μM TPFE nanocomposite carrier, the control group was added with 200μL of PBS, and incubated for 24h under normal cell culture conditions after adding the drug. After 24 hours, the medium containing the drug was aspirated, and 100 μL of CCK-8 working solution diluted 10 times with colorless DMEM was added to each well. The absorbance value of each well was used to calculate the cell viability, the cell viability of each group of cells=(experimental group-blank group) / (contro...

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Abstract

Embodiments of the present invention relate to a nanocomposite carrier for transfection, nucleic acid nanomedicine, a preparation method and application thereof, and belong to the field of biomedicine. The nanocomposite carrier for transfection includes fullerene derivatives and negative The charged high molecular polymer, the fullerene derivative and the negatively charged high molecular polymer are combined through electrostatic interaction and π-π interaction, and the combined nanocomposite carrier has a positive zeta potential, The nanocomposite carrier is combined with siRNA through electrostatic interaction and π-π interaction to obtain nucleic acid nanomedicine. The nanocomposite carrier of the present invention can load siRNA drug, can assist siRNA to pass through cell barrier, improve the transfection performance of siRNA, the formed nucleic acid nanometer drug has stable structure and good biocompatibility, and can effectively protect siRNA from being damaged by RNase. It can be degraded, and the siRNA can be transfected into the lungs by inhalation administration, so that the expression of the target gene in the lung tissue can be effectively inhibited.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a nanocomposite carrier for transfection, a nucleic acid nanomedicine and applications thereof. Background technique [0002] The information disclosed in this Background section is only for enhancement of understanding of the general background of the invention and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person of ordinary skill in the art. [0003] Lung cancer and lung metastases are malignant tumors with the highest incidence in the world, with extremely high recurrence and mortality rates. How to improve the cure rate of lung cancer and lung metastases has become an urgent problem to be solved. At present, the main treatment methods for lung cancer and lung metastases are surgery, chemotherapy, radiotherapy and immune checkpoint blockade therapy, and there are few treatments with sma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K47/52A61K47/59A61K47/61A61K9/72A61K31/713A61K47/64A61P11/00A61P35/00B82Y5/00
CPCA61K47/52A61K47/59A61K47/61A61K47/643A61K31/713A61P35/00A61P11/00A61K9/007B82Y5/00
Inventor 白春礼刘帅舒春英王春儒陈代钦李雪
Owner INST OF CHEM CHINESE ACAD OF SCI
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