Biomimetic multifunctional lipoprotein nanoparticle modified by biological peptide and preparation method and application thereof
A lipoprotein and biological peptide technology, applied in the field of biomimetic multifunctional lipoprotein nanoparticles and their preparation, can solve the problems of difficult to determine the specific process of complex diseases, difficult to achieve disease treatment effects, no clear reports, etc. and drug targeting, improving bioavailability, and avoiding biosafety issues
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Embodiment 1
[0051] Example 1: Preparation of MnAs co-precipitated core and hydrophobic modification
[0052] Take 7.5mL of cyclohexane, 1.5mL of triton, and 1mL of n-hexanol, and mix with magnetic stirring for 10min to form a transparent and stable reverse microemulsion system. Weigh an appropriate amount of manganese chloride tetrahydrate, add 1 mL of ultrapure water, and vortex to dissolve to prepare a manganese chloride solution. Weigh an appropriate amount of ATO, add 1mL of 1M NaOH solution, vortex to dissolve, add concentrated hydrochloric acid, adjust the pH to 7.4, and prepare an ATO solution. 100 μL of manganese chloride solution was added dropwise to the inverse microemulsion system, and magnetically stirred for 15 min. 100 μL of ATO solution with the same molar concentration was added dropwise, and magnetically stirred for 2 hours to form a pale yellow MnAs co-precipitation.
[0053] 100 μL of 20 mmol / L 1,2-oleoylphosphatidic acid sodium salt (DOPA) chloroform solution was ad...
Embodiment 2
[0054] Example 2: Preparation process of carrying MnAs and modifying functional peptide Mel biomimetic lipoprotein nanoparticles (Mel-LNPs / MnAs)
[0055] Weigh 1.5 mg of DMPC, add 0.5 mL of MnAs co-precipitation solution prepared under "Example 1" to dissolve, add dropwise to D4F solution (0.75 mg, 1.5 mL) and stir for 2 h, then add α-Mel solution (0.5 mg, 0.5mL) was stirred and emulsified for 2h. Sonicate (195W) for 10 min, and remove the organic solvent by rotary evaporation. Pass through a 0.8 μm filter membrane, centrifuge ultrafiltration (5000r / min, 10min) with a 30k Da ultrafiltration tube to remove free drug, and resuspend in PBS buffer to obtain Mel-LNPs / MnAs.
[0056]The mass ratio of D4F polypeptide to phospholipid was selected as 0.5:1, and the amount of phospholipid (DMPC / PC) and phospholipid used in the preparation of Mel-LNPs / MnAs was further investigated. After dilution with water, the particle size, polydispersity index (PDI) and Zeta potential of the prepara...
Embodiment 3
[0065] Example 3: Investigation on the Properties of Biomimetic Multifunctional Lipoprotein Nanoparticles Modified by Biological Peptides
[0066] The optimal prescription was selected for the preparation of Mel-LNPs / MnAs, and the following investigations were carried out.
[0067] 1.1 Morphology of Mel-LNPs / MnAs
[0068] Observation with a high-resolution transmission electron microscope showed that figure 2 As shown in A, it can be seen that the shape of the Mel-LNPs / MnAs particles is regular and spherical, and the phenomenon of adhesion and aggregation between the particles is less. After zooming in, it can be seen that there are a certain number of black lattices distributed in the middle of the particles, indicating that the MnAs co-precipitation is better loaded in the LNPs.
[0069] Under the same field of view, the two elements of Mn and As were scanned, and the energy spectrum point scanning test was carried out. The results are as follows figure 2 As shown in B,...
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