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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  • 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 wogo

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

Example Embodiment

[0040] Example 1 Preparation and characterization of DNA tetrahedra

[0041] 1. Method

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

[0043] TDN is self-assembled by four uniquely designed single DNA strands (S1, S2, S3, S4) through a fast, simple and specific PCR program (95℃ for 10min, rapid cooling to 4℃ for 20min, 4℃ for long-term storage) self-assembly Synthetic. The four single strands are added to 96μl of TM buffer (10mMTris-HCl, 50mM MgCl) in an equimolar ratio (1μl of 100μM stock solution for each single strand) 2 , PH 8.0) In a 200μl EP tube, the reaction solution was heated to 95°C for 10 minutes, and then quickly cooled to 4°C to synthesize TDN.

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

[0045]

[0046]

[0047] 1.3 Polyacrylamide gel electrophoresis, dynamic light scattering (DLS), atomic force microscopy (AFM), transmission electron microscopy (TEM) and charge measurement to characterize TDN:

[0048] ①PAGE: The suc...

Example Embodiment

[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 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, and mix so that the concentration of wogonin in the mixed solution is 10μM, 25μM and 50μM, and the TDN concentration is 250nM; then Incubate directly on a constant temperature shaker at 4°C for 8h.

[0059] 2. Characterization of W

[0060] The Zeta potential (Zetasizer Nano ZS90) was measured for W. At the same time, AFM and TEM were used to characterize the morphology and size of W. The result shows that the potential of W is about 17 negative. Combining AFM and TEM results showed uniformly distributed fine particles with a W size of 1-3 nm. ...

Example Embodiment

[0066] Experimental example 1 cell experiment

[0067] 1. Cell type: chondrocytes.

[0068] 2. Experimental settings: When chondrocytes are cultured to generation P1, chondrocytes are digested and cultured in a 6-well plate. After 10% serum is cultured for 12 hours, the gradient is reduced to 8%, 2%, and 1%. After it drops to 1%, do the following:

[0069] Control group: culture for 38h.

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

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

[0072] In the experimental example, "inflammatory cartilage cells" refer to cartilage cells that have been treated with IL-1β (concentration 10ng / ml) for 24 hours.

[0073] 3. Related testing methods and results

[0074] (1) Detect the entry of TDN into normal chondrocytes...

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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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