DNA tetrahedron-wogonin compound and preparation method and application thereof

A technology for wogonin and complex, which is applied in the field of DNA tetrahedron-wogonin complex and its preparation, achieves the effects of improving bone damage, inhibiting inflammatory response, and promising industrialization prospects

Active Publication Date: 2019-04-16
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the anti-inflammatory effect of wogonin is not enough to make wogonin surpass the existing drugs and achieve further substantial therapeutic effect.

Method used

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  • DNA tetrahedron-wogonin compound and preparation method and application thereof
  • DNA tetrahedron-wogonin compound and preparation method and application thereof
  • DNA tetrahedron-wogonin compound and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Preparation and Characterization of Example 1 DNA Tetrahedron

[0041] 1. Method

[0042] 1.1 Preparation method of DNA tetrahedron (TDN)

[0043] TDN is self-assembled by four uniquely designed DNA single strands (S1, S2, S3, S4) through a fast, simple and specific PCR program (95°C for 10min, rapid cooling to 4°C for 20min, long-term storage at 4°C) Synthetic. The four single strands were added to 96 μl TM buffer (10 mM Tris-HCl, 50 mM MgCl 2 , pH 8.0) in a 200μl EP tube, the reaction solution was heated to 95°C for 10min, and then rapidly cooled to 4°C to synthesize TDN.

[0044] 1.2 The specific sequences of the four DNA single strands are as follows:

[0045]

[0046]

[0047] 1.3 Characterization of TDN by polyacrylamide gel electrophoresis, dynamic light scattering (DLS), atomic force microscopy (AFM), transmission electron microscopy (TEM) and charge determination:

[0048] ①PAGE: The successfully synthesized TDN was firstly characterized by 8% polyac...

Embodiment 2

[0055] Example 2 Preparation and characterization of wogonin (W) solution and TDN-wogonin complex (TWC)

[0056] 1. Preparation of W solution and TWC

[0057] Preparation of W solution: W was purchased from Coolaber (China, Beijing), and was directly dissolved in dimethyl sulfoxide (DMSO) to obtain a wogonin solution.

[0058] Preparation of TWC: Take an appropriate amount of wogonin solution, double distilled water (or TM buffer) and TDN solution, mix, so that the concentration of wogonin in the mixed solution is 10 μM, 25 μM and 50 μM, and the concentration of TDN is 250 nM; then Incubate directly on a constant temperature shaker at 4°C for 8h.

[0059] 2. Characterization of W

[0060] Zeta potential (ZetasizerNano ZS90) was measured for W, and the morphology, structure and size of W were characterized by AFM and TEM. The results show that the potential of W is negative about 17. Combining AFM and TEM results show that W is uniformly distributed tiny particles with a si...

experiment example 1

[0066] Experimental Example 1 Cell Experiment

[0067] 1. Cell type: Chondrocytes.

[0068] 2. Experimental setup: when the chondrocytes were subcultured to the P1 generation, the digested chondrocytes were cultured in a 6-well plate, and after 12 hours of culture with 10% serum, they were cultured with 8%, 2%, and 1% serum in gradients. After dropping to 1%, do the following:

[0069] Control group: cultivated for 38 hours.

[0070] Model group: After cultured for 14 hours, IL-1β (concentration: 10 ng / ml) was added to treat for 24 hours.

[0071] Experimental group (TDN, wogonin and TWC): After 12 hours of culture, TDN, wogonin or TWC solution was added for 2 hours, and then IL-1β (concentration: 10 ng / ml) was added to each group for 24 hours.

[0072] In the experimental examples, "inflammatory chondrocytes" refer to chondrocytes treated with IL-1β (concentration: 10 ng / ml) for 24 hours.

[0073] 3. Relevant detection methods and results

[0074] (1) Detection of TDN en...

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Abstract

The invention provides a nano molecule drug-carrying compound, which is formed by encapsulating a DNA tetrahedron and wogonin according to a molar ratio of 1: (40-200). The invention also provides a preparation method of the compound. The invention also provides the application of the compound in a medicament for the treatment of osteoarthritis. Compared with the single use of DNA tetrahedron andwogonin, the compound has better effect in the aspects of inhibiting inflammatory reaction and improving bone injury, and the compound has good industrialization prospect when being used for preparingmedicaments for treating osteoarthritis.

Description

technical field [0001] The invention relates to the field of osteoarthritis drugs, in particular to a DNA tetrahedron-wogonin complex and a preparation method and application thereof. Background technique [0002] Osteoarthritis (OA) is a degenerative disease, which is caused by many factors such as aging, obesity, strain, trauma, congenital abnormalities of joints, joint deformities and other factors. . OA can be divided into primary and secondary types. Primary OA mostly occurs in middle-aged and elderly people, without clear systemic or local inducements, and has a certain relationship with genetic and physical factors. Secondary OA can occur in young adults, and can be secondary to trauma, inflammation, joint instability, chronic and repeated cumulative strain, or congenital diseases. [0003] At present, the drugs for osteoarthritis are mostly anti-inflammatory drugs and analgesics, which can only relieve the pain of patients, and the substantial therapeutic effect i...

Claims

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Application Information

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IPC IPC(8): A61K47/26A61K31/352A61P19/08A61P19/02A61P29/00
CPCA61K31/352A61K47/26A61P19/02A61P19/08A61P29/00
Inventor 林云锋石思容
Owner SICHUAN UNIV
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