Preparation And Quality Control Of Rana Antibacterial Peptide Liposomes

Rana antibacterial peptide is a kind of animal antibacterial peptide with highefficiency and broad-spectrum antibacterial effect extracted from the Rana skin.Studies have found that such antibacterial peptides have a strong inhibitory effect on Gram-positive and negative bacteria,and also have certain anti-inflammatory and analgesic effects.It is a good antibacterial drugs with great development prospects.This experiment is try to design a new formulation of Rana antibacterial peptides on the basis of maintaining the good biological activity,to solve the low bioavailability and short dosing period of Rana antibacterial peptides,and reducing the damage to the human body during the use of Rana antibacterial peptides.Liposomes are artificial biofilms.They are spherical lipid vesicles with a bilayer structure composed of phospholipids.They are not only highly biocompatible,non-toxic,non-immunogenic,and highly stable.It has slow release effect and it is safe and reliable for drug release systems.The Rana antibacterial peptide liposome prepared in this experiment has high encapsulation efficiency,good bacteriostatic effect,long bacteriostatic time,good stability,and high skin retention rate.This article includes the following:1.Rana antimicrobial peptides in vitro establishment of analytical methodology.In this experiment,the in vitro analysis measurement of Rana antibacterial peptides was established by BCA method.The standard curve established has good linearity and high reliability.The repeatability,method recovery and instrument precision of the Rana antibacterial peptides obtained by this method meet the standards for in vitro analysis and determination.The content of antibacterial peptides in the extract of Rana antibacterial peptide was determined by this method,and the determination method of the encapsulation efficiency of Rana antibacterial peptide liposomes was established by combining this method with ultrafiltration centrifugation.2.Preparation and Optimization of Rana Antibacterial Peptide Liposomes.The best preparation technology of Rana antibacterial peptide liposomes was selected from thin-film dispersion,ethanol injection method and thin film homogenization method--thin-film dispersion.The Rana antibacterial peptide liposomes prepared by this method have high encapsulation efficiency,moderate particle size and high stability.Then the effects of egg yolk lecithin content,phospholipid to cholesterol ratio,lipid-drug ratio,and phosphate buffer pH on the encapsulation efficiency of Rana antibacterial peptide liposomes were investigated by single factor analysis.And then screened out three main influencing factors(egg yolk lecithin content,phospholipid-cholesterol ratio,lipid-to-drug ratio)as optimization targets.Using response surface design analysis and fitting model,the best optimal formula for Rana antibacterial peptide liposomes was screened as: The content of egg yolk lecithin was 39.2 mg,the ratio of phospholipid to cholesterol was 3.84 / 1,and the lipid-drug ratio was 4.36 / 1.The theoretical encapsulation efficiency of Rana antibacterial peptide liposome was 79.08%,and the actual encapsulation efficiency was 78.98%.3.Pharmacodynamics of Rana Rana Antibacterial Peptide Liposomes.In the experiment,the bacteriostatic effect of Rana antibacterial peptide liposomes was examined through the bacteriostatic rate and bacteriostatic zone experiments.As a result,the Rana antibacterial peptide liposomes had good bacteriostatic effect and the antibacterial time was longer than Rana antibacterial peptide.Then the skin penetration rate of Rana antibacterial peptide liposome was measured through the transdermal rate experiment.The transdermal rate of Rana antibacterial peptide liposome was calculated to only 6%,and the skin residual rate was high,which effectively reduced the Rana antibacterial peptide Biological toxicity.4.Investigation on the Quality Standard and Stability of Rana Antibacterial Peptide Liposomes.The particle size and potential of Rana antibacterial peptide liposomes was measured by Zetasizer nanometer particle size potentiometer,and the morphology of Rana antibacterial peptide liposomes was observed by Scanning electron microscope.Then conducted influential factor experiments(including temperature,light,humidity,and freeze-thaw),accelerated experiments,and long-term experiments.Influential factor experiments found that Rana antibacterial peptide liposomes exposed to high temperature(60 ° C),high humidity(90%),and under the light conditions,the encapsulation rate and appearance did not change much;subsequent accelerated experiments and long-term experiments found that the appearance state and encapsulation efficiency of Rana antibacterial peptide liposomes did not change much.It shows that the Rana antibacterial peptide liposomes have good stability.