Nano-composite vector for transfection, nucleic acid nano-drug as well as preparation method and application of nano-composite vector and nucleic acid nano-drug

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

Active Publication Date: 2021-08-17
INST OF CHEM CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Nano-composite vector for transfection, nucleic acid nano-drug as well as preparation method and application of nano-composite vector and nucleic acid nano-drug
  • Nano-composite vector for transfection, nucleic acid nano-drug as well as preparation method and application of nano-composite vector and nucleic acid nano-drug
  • Nano-composite vector for transfection, nucleic acid nano-drug as well as preparation method and application of nano-composite vector and nucleic acid nano-drug

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Example 1: siRNA / tetrapiperazine epoxy C 60 Preparation of Derivative / Polythiophene Polymer Nucleic Acid Nanomedicine

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

Embodiment 2

[0048] Embodiment 2: Nano-medicine and the hydration particle size detection of individual components

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

Embodiment 3

[0050] Embodiment 3: CCK-8 method detects the biosafety of nanocomposite carrier

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

[0052] 96-well plate, according to each well 1x10 4 When the confluence reached 40%-50%, add 200 μL of 25 μM, 50 μM, 75 μM, 100 μM, 125 μM, 150 μM of TPFE and 50 μM, 100 μM, 150 μM, 200 μM, 250 μM, 300 μM of PT and containing 25 μM, 50 μM, 75 μM, 100 μM, 125 μM, 150 μM TPFE nanocomposite carrier, the control group was added with 200 μL of PBS, and incubated under normal cell culture conditions for 24 hours after adding the drug. After 24 hours, the medium containing the drug was sucked away, and 100 μL of CCK-8 working solution diluted 10 times with colorless DMEM was added to each well. After incubation for 1 hour under normal cell culture conditions, a multifunctional microplate reader was used to measure t...

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Abstract

Embodiments of the invention relate to a nano-composite vector for transfection, a nucleic acid nano-drug as well as a preparation method and application of the nano-composite vector and the nucleic acid nano-drug, belonging to the field of biomedicines. The nano-composite vector for transfection comprises a fullerene derivative and a high-molecular polymer with negative charges, wherein the fullerene derivative and the negatively-charged high-molecular polymer are combined through electrostatic interaction and pi-pi interaction, the zeta potential of the nano-composite vector formed by combination is positive; and the nano-composite vector and siRNA are combined through electrostatic interaction and pi-pi interaction to obtain the nucleic acid nano-drug. The nano-composite vector can load siRNA drugs, can assist siRNA in penetrating through cell barriers and improves the transfection performance of siRNA; and the formed nucleic acid nano-drug is stable in structure and good in biocompatibility, can effectively protect siRNA from being degraded by RNA enzyme, and achieves lung transfection of siRNA through respiratory tract inhalation drug delivery, and therefore, the expression of a target gene in a 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, nucleic acid nanomedicine and applications thereof. Background technique [0002] The information disclosed in this Background section is only for enhancing the understanding of the general background of the present invention and should not be taken as an acknowledgment or any form of suggestion that the information constitutes the prior art that is already known to those skilled in the art. [0003] Lung cancer and lung metastases are malignant tumors with the highest incidence rate in the world, and the recurrence rate and mortality rate are extremely high. 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 small interfe...

Claims

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Application Information

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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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