Development And Application Of Jiashi Yinchai Tuire Tang

Objective:To study the antipyretic,analgesic,antibacterial and anti-inflammatory effects of extracts from different prescription ratio and different extraction methods of Jiashi Yinchai Tuire Tang,and to realize the verification of prescription ratio and the preliminary exploration of extraction process.The response surface method was used to determine the optimum extraction process of Jiashi Yinchai Tuire Tang from the perspective of multiple indexes and comprehensive evaluation.The optimum forming process of Yinchai Tuire granules was optimized and the pilot production was studied.To formulate the quality standard(draft)of Yinchai Tuire granules.Carry out quality inspection and stability investigation on pilot products.To study the pharmacologic equivalence of Yinchai Tuire granule and Yinchai Tuire oral liquid,so as to provide reference for clinical rational drug use.To investigate the mechanism of Jiashi Yinchai Tuire Tang in the treatment of acute upper respiratory tract infection by means of network pharmacologyMethods:1.To investigate the effects of Jiashi Yinchai Tuire Tang 1-6 on the activity of staphylococcus aureus,the rat model of fever caused by dry yeast,the rat model of foot swelling caused by egg white and the writhing reaction caused by acetic acid in mice.Test samples 1-3:the ratio of Lonicerajaponica Flos and Bupleuri radix was 1:1,2:1 and 1:2,and the extraction method is water decoction.Test samples 4～6:the ratio of Lonicerajaponica Flos and Bupleuri radix was 1:1,2:1 and 1:2,and the extraction method was double formulation2.HPLC method was used for quantitative analysis,with the transfer rate of chlorogenic acid and baicalin as the response value,and the box-behnken Design(BBD)mode in response surface method,to optimize the extraction process parameters of Jiashi Yinchai Tuire Tang,such as boiling time(A),boiling times(B)and water content(C)3.The optimum preparation process of traditional wet granulation was screened by the parameters of molding rate,solubility and resting Angle.The amount,temperature and ratio of auxiliary materials were optimized by taking the molding rate as the index.Based on the results of pilot production,the forming process of Yinchai Tuire granules was established,and the main technical parameters and the types and dosage of auxiliary materials were determined4.The quality standard of antipyretic granules was studied from four aspects identification,fingerprint,inspection(preparation general inspection,arsenic salt inspection,heavy metal and harmful elements inspection,microbial limit inspection)and content determination.The identification includes character and TLC identification.HPLC method was used to establish the method for the determination of fingerprints.The similarity evaluation system for the similarity evaluation of 11 batches of Yinchai Tuire granules was used to evaluate the similarity of the fingerprints of 11 batches of Yinchai Tuire granules According to the requirements of "granules" in the fourth general rule of”Chinese pharmacopoeia"(2015 edition),the particle size,water content,drying weight loss,solubility,loading quantity difference and microbial limit of antipyretic particles of silver firewood were examined.Arsenic salt was examined by Gutzeit Test.The detection of heavy metals was carried out by sodium colorimetry.The harmful elements Cu,Pb and Cd were determined by atomic absorption spectrophotometry,Cu by flame method,Pb and Cd by graphite furnace method.The effective components of honeysuckle,forsythia,radix Scutellariae and radix bupleuri were determined by HPLC5.According to the quality standard(draft),Under the condition of commercially available packaging,the 6-month stability test and acceleration test were carried out on three batches of pilot test products of Yinchai Tuire granules[the acceleration test conditions were temperature(40±2℃)and humidity(75±5%),and the long-term stability test conditions were temperature(25±2℃)and humidity(60±10%)]6.Through in vitro antibacterial experiment,using staphylococcus aureus as the test strain,the antibacterial effects of Yinchai Tuire granules and oral liquid were compared;The antipyretic effects of Yinchai Tuire granules and oral liquid were compared by dry yeast-induced rat model.The anti-inflammatory effects of y Yinchai Tuire granules and oral liquid were compared by the models of egg white induced swelling of rats’ feet and ear swelling of mice caused by xylene.The analgesic effects of Yinchai Tuire granules and oral liquid were compared by means of writhing reaction induced by acetic acid in mice and hot plate method7.Collect the active ingredients related to the medicinal taste contained in the prescription of Jiashi Yinchai Tuire Tang through the pharmacology database of traditional Chinese medicine system(TCMSP).Swiss Target Prediction database(Swiss Target Prediction)Acute upper respiratory tract related targets were obtained through the treatment target database(TTD),Drugbank,and DisGeNET.The Venn diagram obtains the relevant intersection.The target protein interaction was analyzed using the STRING website,and the results were visualized using Cytoscape 3.7.1.Gene ontology(GO)analysis and KEGG pathway enrichment analysis were performed using DAVID databaseResults:1.In each experiment,the differences between the Tests 1-6 and the model group were statistically significant(P<0.05).Jiashi Yinchai Tuire Tang extracts with different prescription ratios and different extraction processes all have certain antibacterial,antipyretic,anti-inflammatory and analgesic effects.When Lonicerae Japonicae Flos:Radix Bupleuri 1:1 and the extraction process is steam distillation+water boiling,the antibacterial,antipyretic,anti-inflammatory and analgesic effects are more obvious2.A method was established for the determination of chlorogenic acid and baicalin in the extract of Jiashi Yinchai Tuire Tang.The optimal extraction conditions of Jiashi Yinchai Tuire Tang were as follows:boiling time 60min,boiling twice,adding 12 times of water3.The best techniques for traditional dry granulation were:the dosage of extract powder and auxiliary material was l:0.3,the auxiliary material was icing sugar:dextrin was 2:1,the dosage of aspartame was 1%,and the appropriate amount of ethanol was 90%.The optimal spray pelletizing process was 1.20 relative density of the extract,90-40 inlet temperature-outlet temperture,10 Hz spray rate,sugar powder:dextrin ratio was 1:1.Less spray pelletizing process,the whole process of pelletizing is closed operation,less dust,is conducive to reduce the loss of effective ingredients,more in line with GMP standards,and at present,most pharmaceutical factories use spray pelletizing equipment,so the pelletizing method is determined as spray pelletizing4.This product is yellowish-brown,taste slightly sweet,slightly bitter.In the TLC identification,the TLC identification method of 9 drugs including radix bupleuri,radix Scutellariae,honeysuckle,forsythia,radix purpura,lysimachia leaf,licorice root,rhizoma cyperi,and burdock was determined.In the fingerprint study,12 common peaks were identified and 8 common peaks were identified,which were peak 1 neocorbic acid,peak 2 chlorogenic acid,peak 4 cryPtochlorogenic acid,peak 7 forsythioside A,peak 8 isochlorogenic acid B,peak 9 isochlorogenic acid C,peak 10 baicalin and peak 12 baicalin The test results of particle size,water content,drying weight loss,solubility,load difference and microbial limit of Yinchai Tuire granules all meet the requirements under "granules" of general rule 0104 of the fourth part of the 2015 edition.The inspection limits of arsenic salt and heavy metal were less than 3ppm.The average contents of each element in the inspection of harmful elements were Cu 1.83mg/kg,Pb 0.32mg/kg and Cd 0.22mg/kg.A multi-index HPLC method was established for the determination of Yinchai Tuire granules.Finally,it was determined that each bag of Yinchai Tuire granules contained honeysuckle and arctium dock seeds with chlorogenic acid(C16H18O9),and the content should not be less than 6.3mg.Forsythia with forsythia ester glycoside A(C29H36O15),not less than 11.8 mg;Containing baicalin(C21H18O11),not less than 40.0 mg.The quality standard of antipyretic granules of silver firewood(draft)was established5.The appearance,characteristics,identification,solubility,moisture content,particle size,and content determination of Yinchai Tuire granules all meet the quality standard(draft)6.The differences between the model group and the Yinchai Tuire granules and oral liquid group were statistically significant(P<0.05).There was no significant difference between Yinchai Tuire granules and oral liquid group,which had pharmacological equivalence7.There were a total of 322 active components of traditional Chinese medicine in Jiashi Yinchai Tuire Tang,corresponding to 1088 different targets,and 623 targets related to acute upper respiratory tract infection were collected.Jiashi Yinchai Tuire Tang mainly ACTS on TNF,toll-like receptor,nod-like receptor,MAPK and T cell receptor signaling pathways,which are mainly involved in inflammation and immune regulation.Conclusion:The extracts of Jiashi Yinchai Tuire Tang with different prescription ratio and different extraction techniques have different antibacterial,antipyretic,anti-inflammatory and analgesic effects When the ratio of Lonicera japonica Flos and Bupleuri radix was 1:1,and the extraction process was double,the antibacterial,antipyretic,anti-inflammatory and analgesic effects were more obvious.The optimized extraction process and molding process are stable,feasible and operable,and suitable for mass production.The quality standard(draft)of Yinchai Tuire granules was established and its stability was investigated.Both Yinchai Tuire granules and oral liquid have different degrees of antibacterial,anti-inflammatory,antipyretic and analgesic effects.In the treatment of acute upper respiratory tract infection,Jiashi Yinchai Tuire Tang function correlation between pathway,removing inflammation,restrain the activation of the role of cytokines,alleviating ultra immune response,and the traditional Chinese medicine compound of corresponds to the overall regulatory characteristics,embodies the Chinese medicine composition more-the role of multiple targets-channel characteristics.