Researchon Preparation Of The Sophora Flavescens Ait. Alkaloids Transfersome Gel And Their Clinical Application

Bovine mastitis is one of the three major diseases which was harmful to the development of dairy farming seriously.It has a high incidence,widespread epidemic,strong contagion and harmfulness.Therefore,effective control of bovine mastitis plays an important role in improving dairy production performance,reducing economic loss and promoting continuous healthy development of dairy industry.At present,the most commonly used therapeutic regimen was antibiotic treatment.This method is effective in the initial stage of application,but long-term application leads to drug residue,bacterial resistance and relapse.With the increasing awareness of food and drug safety and the rapid upgrading of modern separation and purification technology of traditional Chinese medicine efficacy components,the natural products of traditional Chinese medicine have won the public's recognition with its high cost performance and safety.Conventional methods of administration,such as oral and injection,often cause adverse reactions such as gastrointestinal stimulation,liver first-pass effect,poor compliance,difficult to use and other problems.Percutaneous administration is the more direct,effective,safe and convenient way to treat dairy cow mastitis,but it is difficult to break and through the barrier of cuticle and to improve the efficiency of drug percutaneous absorption,which limits its application.No Chinese herbal extract of cow mastitis was found on the market and in the field of scientific research.So far,The transdermal drug type which used in dairy mastitis has not been found on the market or in the field of scientific research.The purpose of this study was to provide a stable performance,high security and effectiveness preparation for the treatment of dairy cow mastitis in clinical practice.The research has been prepared a SFAA transfersome gel with antibacterial,anti inflammatory,anti swelling,analgesic,antiviral and antifungal functions for the treatment of dairy cow mastitis.The physical and chemical properties,pharmacological activities,stability and safety of the preparation,as well as the clinical efficacy of this preparation were tested.This study provides a reference for the development of new drugs in prevention and treatment of cow mastitis,also provides data support for clinical medication of it.In this study,high performance liquid chromatograph?HPLC?was used to establish an analytical method for the determination of matrine?MAT?and Oxymatrine?OMT?in the prepa-ration.The prescription and process parameters of the SFAA transfersome were optimized by the two regression orthogonal rotation combination design.Physical and chemical properties of the SFAA transfersome were tested.The micromirror was used to observe the appearance character,the particle size and Zeta potential were detected by the laser particle size analyzer,the deformability was evaluated by measuring the relative permeable rate,and the embedding ratio of the SFAA transfersome was measured by the HPLC which established by the application.The preparation parameters of the SFAA transfersome gel were optimized by the response surface method.The release properties and transdermal absorption effects were evaluated by the cumulative release degree,the cumulative permeability,the residual amount of the skin and the steady infiltration rate in vitro,and the inhibitory effects on the common pathogens causing dairy cow mastitis were evaluated by MIC and MBC test.The stability was tested by the viscosity,relative permeable rate and encapsulation rate,the safety was evaluated by skin irritation test,allergy test and acute toxicity test.The clinical contrast test was carried out with the SFAA transfersome gel and Wan Ru kang for the recessive mastitis,and with SFAA transfersome gel and penicillin for the clinical mastitis.The results of the related research are as follows:A HPLC detection method for determining the content of main drug components in SFAA transfersome was established:the calibration curves of MAT and OMT were in good linearity over the rang of 0.074⁰.370?g·mL^-11 and 0.263¹.315?g·mL^-1,respectively.The method has good reproducibility,high precision and high recovery rate,can be used for determine the embedding ratio of SFAA transfersome.The optimal formulation for preparing SFAA transfersome gel was phospholipid mass concentration of 2.75%,phospholipid mass ratio of 3:1,1,2-propylene glycol 0.5%,hydration time 34.4 min;Transmission electron microscopy?TEM?observation showed that:the shape of their particles was spherical or similar to spherical under microscope,which was smooth with an average diameter of 184.2±2.7 nm,Zeta potential of-47.4±2.3 mV,and a through membrane ratio relative of 76.09±0.34%,RSD=1.00%,n=3.The embedding ratio of MAT and OMT,which comprised in SFAA transfersome,were 80.43±0.98%and 74.70±1.08%respectively.The optimal proportion of matrix components was made from carbomer 0.68%,azone 3.52%,and glycerine3.56%.It shows that the response surface analytical method is reliable and can be used in the process of prescription optimization,and the optimal for mulation of SFAA transfersome gel based the optimal for mulation is well distributed and transparent,it has proper consistency and well rheopexy.Compared with the common gel group,the SFAA transfersome gel group was 2.22 times the rate of cumulative transmission at 24 h?p<0.05?,it was 5.27 times the rate of retention in the skin layer?p<0.05?,and it was 1.51 times the rate of steady state penetration rate.The MICs of SFAA to E.coli,S.aureus,S.agalactiae and S.dysgalactiae,were 3.13 mg·mL^-1,6.25 mg·mL^-1,0.78 mg·mL^-1,0.78 mg·mL^-11 respectively,and the MBCs were 6.25 mg·mL^-1,6.25 mg·mL^-1,1.56 mg·mL^-1,0.78mg·mL^-1,respectively.The SFAA transfersome gel had effective transdermal absorption and antibacterial effects on the 4 kinds of bacterial pathogens of cow mastitis.The SFAA transfersome gel can stable storage under 4?or 25?at room temperature until180d,and the embedding ratio drops quickly under 40?,29.85 percents was descended in 180 d.The results showed that this preparation under the condition of low temperature of 4?or room temperature 25?had a high stability,can deposit 180d,the quality can be controlled.The cutaneous stimulation test showed that the average score,which displayed skin irritation degree,of the high doses gel was 0.375<0.4,and the other groups were under 0 all.It means that applied SFAA transfersome gel of low,middle,or high dose in the rats,there were no local erythema,edema and irritation no matter in intact-skin and damaged-skin,and with no statistically significant difference compared with the control group?P>0.05?.The hypersensitivity test showed that the sensibilization rate of the gel was 0.And all of the rats had no death and obviously toxic symptoms,the body mass had an normal increase,there was no statistically significant difference of body mass between the blank groundmass group and observation group?P>0.05?.The gel of different dose has no acute toxicity,cutaneous stimulation and irritability in experimental animals.It was safety for external medication.The SFAA transfersome gel on mastitis cure rate was 86.67%,the effective rate was 100%.The cure rate is higher than the control group Wan Ru Kang 20 percentage points;according to clinical mastitis,respectively by SFAA transfersome gel and mycillin clinical application of comparative test,the results showed that SFAA transfersome gel on clinical mastitis has a good effect,the cure rate was 73.33%,efficiency was 93.33%,the cure rate is higher than the control group mycillin 6.66 percentage points.Quality standard formulation of SFAA transfersome:Appearance should be spherical or ellipsoid,smooth and not sticky.The average particle size should between 180 nm and 190 nm,the Zeta potential should between-42 mV and-50 mV,and the encapsulation efficiency should between 72%and 77%;Quality standard formulation of SFAA transfersome gel:The appearance characters should be white,translucent,uniform fine,a slightly thicker colloidal preparation.The pH should be 6.0⁷.0,the viscosity should be 45⁴6 Pa·s,and the microbial limit:the number of bacteria should be less than 100 cfu·g^-1,mold and yeast count should be less than 100 cfu·g^-1,and staphylococcus aureus and pseudomonas aeruginosa should be not detected.