Nano defensin gel scaffold as well as preparation method and application thereof
A defensin and nanotechnology, which is applied in the direction of pharmaceutical formulas, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the problems of nano-silver/silver ion toxicity, unsuitable for large-scale use, etc., to promote the growth Migration of fibroblasts, ease of drug application, and accelerated wound repair
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0036] Example 1 Synthesis of Human Alpha Defensin 5 (HD5)
[0037]HD5 was synthesized in an Applied Biosystems 433A peptide synthesizer by tert-butoxycarbonyl (Boc) solid-phase peptide synthesis. First, 2.2 mmol of Boc amino acid was added to DMF for activation for 3 min, and then slowly added to 0.25 mmol of PAM resin for coupling for 10 min. TFA (100%) was used to remove the N-Boc group throughout the synthesis, DMF and DCM were used as batch detergents. After the peptide chains were assembled on the BOC resin, anhydrous HF was added to incubate at 0°C for 1 h to remove the protective group, and then added to cold ether for precipitation. Then, one-step renaturation was used to make the HD5 peptide chain form three pairs of key disulfide bonds, and 3 mM reduced glutathione, 3 mM oxidized glutathione and different concentrations of GuHCl were added to the crude peptide system. Simultaneously quickly add NaHCO 3 Adjust the final pH to 8.3. Final overnight incubation to fo...
Embodiment 2
[0038] The synthesis of embodiment 2 nano defensins (ND)
[0039] The peptide chain was assembled and synthesized according to the method described in Example 1. After the peptide chain was assembled, the Fmoc protecting group on the N-terminal lysine residue was eluted with a DMF solution containing 20% piperidine, and then in Applied Biosystems In the 433A polypeptide synthesizer, myristic acid (myristic acid) was coupled to the ε-amino group of lysine according to the same coupling strategy as described in Example 1. Add 25% N,N-dimethylformamide, 2M urea, 50mM Tris / HCl, 3mM reduced glutathione and 3mM oxidized glutathione to the crude peptide chain at 0.25mg / ml, and quickly add NaHCO 3 Adjust the final pH to 8.3 and incubate overnight to naturally oxidize to form three pairs of disulfide bonds. Finally, it was purified by reverse-phase high-performance liquid chromatography (RP-HPLC) and then freeze-dried to obtain the ND product powder.
Embodiment 3
[0040] Example 3 Preparation of nano defensin gel scaffold
[0041] Preparation process and formulation design such as figure 1 As shown, the [PEO] x -[PPO] y -[PEO] z The tri-block copolymer was mixed with sterilized water in an ice bath according to 18% (w / v), and then placed in a 4°C environment for full swelling for 12h; after swelling, placed in a -20°C environment for 5min, and then slowly stirred for 30s. Sonicate in an ice bath until the triblock copolymer dissolves to form a clear and transparent solution, add nano-defensins to make the system concentration 50 μg / ml, and mix thoroughly to obtain nano-defensins gel scaffold (50 μg / ml, 18%).
PUM
| Property | Measurement | Unit |
|---|---|---|
| particle size | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More