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MiRNA (micro Ribonucleic Acid) vector based on lanthanide oxygen-fluorine nanocrystal and application of miRNA vector

A nanocrystal and carrier technology, applied in the field of miRNA carrier and drug carrier materials based on lanthanide oxyfluoride nanocrystals, can solve the problems of poor therapeutic effect, poor biocompatibility, and insufficient miRNA binding ability, etc., and achieve enhanced effects , efficient loading, and good biocompatibility

Active Publication Date: 2022-05-24
陕西万佳态元生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a miRNA carrier based on lanthanide oxyfluoride nanocrystals, which can be used to load miRNA to solve the problems of poor biocompatibility, insufficient miRNA binding ability, and miRNA intracellular transport in miRNA clinical treatment drugs. Insufficient ability, poor actual treatment effect, etc.

Method used

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  • MiRNA (micro Ribonucleic Acid) vector based on lanthanide oxygen-fluorine nanocrystal and application of miRNA vector
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  • MiRNA (micro Ribonucleic Acid) vector based on lanthanide oxygen-fluorine nanocrystal and application of miRNA vector

Examples

Experimental program
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Effect test

Embodiment 1

[0034] This example provides the preparation and characterization of LnP.

[0035] Preparation of LnP: The synthesis of Ln was free oleylamine GdOF: 45% Ce, 15% Tb, the prepared Ln was redispersed in PBS (pH=7.4) buffered saline. The PLL and LDN were coupled by a coordinate bond through the amino group in the polymer. 5 mg of Ln were dissolved in 10 mL of PBS buffer containing 10 mg of PLL. After incubation at 30°C for 60 min with stirring, LnP was collected by centrifugation at 10,000 g.

[0036] Characterization of LnP: The chemical structures of Ln, PLL and LNP were analyzed by infrared spectroscopy. The morphology was observed by transmission electron microscope. Zeta potential and hydrodynamic dimensions were determined with a laser particle sizer. Fourier transform infrared spectroscopy (FTIR) confirmed the coordination bond between PLL and Ln, 1659cm -1 This is also proved by the obvious reduction of the characteristic infrared absorption peaks of amino groups (see...

Embodiment 2

[0038] This example provides the preparation and characterization of LnP-loaded miR-30c.

[0039] The temperature was 37 °C, miR-30c was electrostatically adsorbed for 30 min, and miR-30c was loaded onto the nanoparticle LnP to form a nanoparticle / miR30c complex.

[0040] Preparation of miR-30c complexes: Before preparing miR-30c complexes, miR-30c was centrifuged at 12000 rpm for 10 min and dissolved in fresh DEPC water. LnP and PLL solutions were added to an equal volume of miR-30c solution (0.264 mg / mL), and the miR-30c complexes with different specific gravities were calculated. The miR-30c complex was prepared in a HEPES environment (PH=6.84) at 37°C. The particle size and Zeta potential of various miR-30c complexes were measured by hydrodynamic particle size distribution DLS. All complexes were prepared in 1 mL of ultrapure water containing 2.6 μg miR-30c (w / w=12.5 w / w) for differential scanning calorimetry analysis.

[0041] Characterization of LnP-loaded miR-30c: Th...

Embodiment 3

[0046] This example provides a verification test that LnP efficiently transports MiR-30c in cells.

[0047] The cellular uptake of LnP / miR-30c, PEI25K / miR-30c and Lipo2000 / miR-30c was assessed by monitoring the green fluorescence of FAM-labeled miR-30c on a confocal laser microscope (CLSM, FV1200, Olympus) (see Figure 4 ). In order to screen the optimal weight ratio of LnP:miR-30c for efficient internalization of cells, this example first tested the cellular uptake of LnP / miR-30c at different weight ratios (50:1-400:1). Laser confocal images (see Figure 4 A) and the green fluorescence intensity analysis shown (see Figure 4 B), LnP / miR-30c with a weight ratio of 300:1 had the strongest cellular internalization. In addition, LNP / miR-30c (weight ratio 300:1) also showed the brightest green fluorescence in MC38 cells compared to Lipo2000 / miR-30c and LNP / miR-30c (see Figure 4C). These results suggest that LnP is an efficient miR-30c vector and an optimal alternative to com...

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Abstract

The invention provides a miRNA (micro Ribonucleic Acid) carrier based on lanthanide oxygen fluorine nanocrystals and application of the miRNA carrier. The invention discloses a miRNA (micro Ribonucleic Acid) carrier based on lanthanide oxyfluoride nanocrystals, and an application of the carrier loaded anti-tumor miRNA as a miRNA therapeutic drug. According to the miRNA carrier based on the lanthanide oxygen fluorine nanocrystal, provided by the invention, efficient loading of miRNA can be realized, and miR-30c can be protected in cells for more than 24 hours. The carrier loaded with the anti-tumor miRNA is used as a miRNA treatment drug, so that the miRNA function is reserved, and meanwhile, the carrier still has good biocompatibility.

Description

technical field [0001] The invention belongs to the technical field of nano-composite materials, and relates to a drug carrier material, in particular to a miRNA carrier based on lanthanide oxygen-fluorine nanocrystals and its application. Background technique [0002] The Wnt signaling pathway plays a crucial role in embryonic development, tissue homeostasis, and stem cell proliferation and differentiation in all animals. However, aberrant activation of this signal underlies a variety of human cancers, and dysregulation of the Wnt signaling pathway allows the transcriptional activator β-catenin to accumulate in the cytoplasm and nucleus, thereby promoting the expression of genes involved in tumor cell proliferation and migration. In this process, β-catenin co-activators (including Bcl9), as part of the abnormal activation of Wnt signaling pathway, are overexpressed in a variety of malignant tumors, and form stable complexes with β-catenin to promote these oncogenes express...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/02A61K47/62A61K31/7105A61P35/00
CPCA61K47/62A61K47/02A61K31/7105A61P35/00Y02A50/30
Inventor 闫瑾尤伟名
Owner 陕西万佳态元生物科技有限公司
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