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AuNPs/miR-140 compound as well as preparation method and application thereof

A compound and pharmaceutical preparation technology, applied in the field of medicine, can solve the problems of increasing the risk of cardiovascular disease, indigestion, side effects, etc., and achieve the effects of inhibiting inflammation, repairing cartilage damage, and promoting cartilage repair

Active Publication Date: 2020-08-14
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the side effects are relatively small compared with other NSAIDs, the inventors found that Lnxc can still cause gastrointestinal side effects after oral administration, including gastrointestinal pain, indigestion, nausea, vomiting, etc.
In addition, according to the FDA notification, taking NSAIDs can also increase the risk of cardiovascular diseases, such as myocardial infarction and stroke

Method used

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  • AuNPs/miR-140 compound as well as preparation method and application thereof
  • AuNPs/miR-140 compound as well as preparation method and application thereof
  • AuNPs/miR-140 compound as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Example 1 Preparation of AuNPs

[0084] Add 20 mL triple distilled water into the round bottom flask, and then add 10 mg / mL HAuCl to it under the condition of stirring in a water bath at 60 °C 4 solution 400 μL, then add 800 μL PEI aqueous solution with a concentration of 3 mg / mL, and react for 1 h to obtain AuNPs.

[0085] The effect of PEI volume on the preparation of AuNPs: Add 20 mL triple distilled water to the round bottom flask, and then add 10 mg / mL HAuCl to it under the condition of stirring in a water bath at 60 °C 4 Solution 400μL, and then add 100μL, 200μL, 400μL, 800μL, 1.6mL, 3.2mL, 6.4mL of PEI aqueous solution with a concentration of 3mg / mL, react for 1h, and finally dilute the obtained solution by 2 times and then conduct wavelength analysis at 300-700nm. Scanning, according to the characteristic curve of ultraviolet-visible absorption of different solutions, determine the optimal PEI volume.

[0086] Fix the reaction temperature and time constant, ...

Embodiment 2

[0101] Example 2 Preparation of gold nanoparticles / miR-140 complexes

[0102] Connect miR-140 to AuNPs by electrostatic adsorption. After mixing different concentrations of miR-140 solutions and AuNPs solutions, immediately vortex for 30s to mix well, and then incubated for 30min to obtain different mass ratios of AuNPs / miR- 140 complexes.

[0103] 1. Screening for the optimal mass ratio of the complex (W / W=Au / miR-140)

[0104] Agarose gel retardation electrophoresis: The ability of AuNPs to compress miR-140 was investigated by agarose gel retardation electrophoresis experiments. Accurately weigh 0.2g of agarose and place it in a 50mL Erlenmeyer flask, and add 20mL of 1×TAE electrophoresis buffer to it, heat the Erlenmeyer flask in a microwave oven until the agarose boils and dissolves, and wait for the agarose to cool to 50-65°C Then add 2 μL type Ⅰ nucleic acid staining agent to the triangular beaker and mix well, then gently pour the agarose into the horizontal groove w...

Embodiment 3

[0114] Example 3 In vitro evaluation of AuNPs / miR-140 complexes

[0115] The complex of AuNPs / miR-140 must have small cytotoxicity and high cell transfection ability to play the gene regulation role of miR-140, thereby exerting the effect of treating OA. In this example, chondrocytes were cultured in vitro, and then the cytotoxicity, cell transfection ability and in vitro gene regulation effect of the AuNPs / miR-140 complex were investigated by MTT assay, cell transfection experiment and qPCR experiment.

[0116] Reagents and Instruments

[0117] cy3-miR-140 (Shanghai Jima Pharmaceutical Technology Co., Ltd.); Tetramethylazozolium salt (MTT, Sigma, USA); DMSO for cell culture (Beijing Soleibao Technology Co., Ltd.); 4% paraformaldehyde ( Beijing Suolaibao Technology Co., Ltd.); DAPI (Beijing Suolaibao Technology Co., Ltd.); GAPDH, COL2 primers, U6, miR-140 primers (Shanghai Jima Technology Pharmaceutical Co., Ltd.); TRNzol Universal reagent (Tiangen Biochemical Technology ...

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Abstract

The invention provides an AuNPs / miR-140 compound as well as a preparation method and application thereof. The AuNPs / miR-140 compound is obtained by compounding gold nanoparticles AuNPs and miR-140 through electrostatic adsorption, wherein the mass ratio of the AuNPs to the miR-140 is (5-28.8):1. The AuNPs / miR-140 compound can efficiently deliver the miR-140 into cartilage cells, the cell transfection rate is close to 100%, the AuNPs / miR-140 compound has no obvious toxicity to the cartilage cells, the expression of the miR-140 in the cartilage cells can be up-regulated, the expression level ofCOL2A mRNA in the cartilage cells can be increased, and the AuNPs / miR-140 compound has a repairing effect and a certain anti-inflammatory capability on the cartilage cells. A pharmaceutical composition of the AuNPs / miR-140 compound and lornoxicam has remarkable effects in the aspects of protecting cartilage, promoting cartilage repair and preventing and treating arthritis diseases.

Description

technical field [0001] The invention relates to the field of medicine, in particular to an AuNPs / miR-140 complex 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 present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Osteoarthritis (OA) is a common degenerative disease of articular cartilage. Obesity, aging, strain, trauma, congenital abnormalities of the joints and many other factors can cause the degeneration and loss of articular cartilage, which in turn leads to the regeneration of joint margins and subchondral bone and triggers synovial inflammation. OA can cause joint pain, deformity, limited mobility and muscle wasting in patients, and has become one of the most common disabl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/52A61K47/58A61K31/7105A61K48/00A61K31/542A61P19/02A61P19/04A61P19/08A61M5/20A61M5/31
CPCA61K31/542A61K31/7105A61M5/20A61M5/31A61M2005/2006A61K47/52A61K47/58A61P19/02A61P19/04A61P19/08A61K2300/00
Inventor 黄桂华杨雪华黄小雨
Owner SHANDONG UNIV
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