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A kind of protonated morpholine modified phosphorus-containing dendrimer and its preparation and application

A dendrimer, morpholine technology, applied in biochemical equipment and methods, bone/connective tissue cells, prostheses, etc., to achieve the effects of simple preparation process, easy operation and good application prospects

Active Publication Date: 2020-10-27
DONGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is no report on the use of phosphorus-containing dendrimers to promote osteogenic differentiation of mesenchymal stem cells

Method used

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  • A kind of protonated morpholine modified phosphorus-containing dendrimer and its preparation and application
  • A kind of protonated morpholine modified phosphorus-containing dendrimer and its preparation and application
  • A kind of protonated morpholine modified phosphorus-containing dendrimer and its preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Weigh the second generation phosphorus-containing dendrimer G2-P(S)Cl 2 20mg (4.19μmol) was dissolved in 1mL of anhydrous tetrahydrofuran (THF), and 43.85μL (251.73μmol) of N, N-diisopropylethylamine (DIPEA) was added under the condition of ice bath under nitrogen protection, and then slowly dropped Add 16.51 μL (125.86 μmol) of 4-(2-aminoethyl)morpholine. The ice bath was removed 5 minutes after the dropwise addition was completed, and stirring was continued at room temperature for 3 hours. The reaction solution was spin-dried by a rotary evaporator, and the residual substance was dissolved in 1-2 mL of DMSO to obtain a crude product solution containing G2-Mor. The above liquid was transferred to a reproducible dialysis bag with a molecular weight cut-off of 1000, dialyzed in phosphate buffer solution (pH=7.4) for one day (2L×3) and distilled water for two days (2L×6), and then freeze-dried Purified samples were processed and stored at 4°C.

[0050] See attached ...

Embodiment 2

[0053] The morpholine-modified phosphorus-containing dendrimer (G2-Mor) prepared above was dissolved in anhydrous tetrahydrofuran, and a diethyl ether solution (0.1 M) of hydrochloric acid was added dropwise until no precipitation occurred. The precipitate was obtained by filtration and washed with ether to remove excess hydrochloric acid, and dried in a vacuum oven to obtain the purified protonated product G2-Mor + .

Embodiment 3

[0055] Bone marrow mesenchymal stem cells BMSCs were used as model cells to test the cytotoxicity of the materials prepared in Example 2. Take 1.5×10 4 BMSCs were planted in a 96-well plate and cultured in 100 μL DMEM (L) culture medium supplemented with 100 U / mL penicillin, 100 U / mL streptomycin and 10% FBS at 37 °C and 5% carbon dioxide concentration Cultivate for 24h. Then the medium was changed to G2-Mor + Serum-free medium with concentrations of 0nM, 50nM, 100nM, 500nM, 1000nM, 2000nM and 3000nM were co-cultured with cells for 24h, then washed with PBS, then added 10μL of CCK-8 solution and 90μL of complete medium, and continued to culture for 2h. Finally, use a multifunctional microplate reader to test the absorbance value, and the test wavelength is 450nm. The results are as attached to the instruction manual. figure 2 shown. The results showed that with the increase of the material concentration, the material had almost no effect on the viability of the cells, indi...

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Abstract

The present invention relates to a protonated-morpholine-modified phosphorus-containing dendrimer and preparation and application thereof, morpholine is grafted to the surface of the phosphorus-containing dendrimer, and then protonated to obtain the protonated-morpholine-modified phosphorus-containing dendrimer. The morpholine-modified phosphorus-containing dendrimer is a good reagent for promoting the osteoinductive differentiation of mesenchymal stem cells, and has a good application prospect in bone tissue repair and reconstruction and other aspects.

Description

technical field [0001] The invention belongs to the field of functional dendrimer complexes and their preparation and application, in particular to a phosphorus-containing dendrimer modified by protonated morpholine and its preparation and application. Background technique [0002] Diseases such as bone injury have become a serious global public health problem. According to the degree of bone injury, it can be divided into mild bone injury and large bone defect. For the repair of large bone injuries, bone tissue engineering methods have emerged as the times require. In recent years, stem cells have gradually become a hot spot in bone tissue engineering research. Stem cells are a class of cells that can self-renew and have multilineage differentiation potential. According to the different development time, stem cells can be divided into embryonic stem cells and adult stem cells, and adult stem cells include mesenchymal stem cells (MSCs, mesenchymal stem cells) and other ste...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G83/00C12N5/077A61L27/54A61L27/38A61L27/18
CPCA61L27/18A61L27/3834A61L27/3847A61L27/3895A61L27/54A61L2300/412A61L2430/02C08G83/004C12N5/0654C12N2500/30C12N2506/1353C08L87/00
Inventor 曹雪雁李爱军范钰陈亮史向阳王乐
Owner DONGHUA UNIV
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