Preparation And Efficacy Evaluation Of Enteric-coated Pellets Of Anti-hepatic Fibrosis Active Site From Periplaneta Americana

ObjectiveThe previous research of the research group found that the extract of Periplaneta americana has the anti-hepatic fibrosis effect.This study uses the anti-hepatic fibrosis active site（PA-B）of Periplaneta americana as the raw material to prepare enteric pellets and gastric pellets（not wrapped the enteric coating film）,and uses liver fibrosis rats to investigate the effects of gastric and enteric absorption on drug efficacy and thus laid a foundation for the research and development of the anti-hepatic fibrosis drug of Periplaneta americana.Method1.Using the Central Composite Design of three factors and five levels of response surface method,through the overall evaluation of molding rate and moisture-absorption rate as the evaluation index,the process of optimizing the proportion of moisture resistance excipients is improved.The active site pellets of Periplaneta americana were prepared by the coating pan method,and wrapped the enteric coating film of eudragit L-100 to determine the physical indexes and cumulative releasing of pellets.The method of releasing refers to the regulations of the Chinese Pharmacopoeia 2015Releasing.The enteric properties of the pellets were determined by in vitro release and preliminary targeting evaluation in vivo.2.The models of liver fibrosis induced by intraperitoneal injection of carbon tetrachloride were randomly divided into model group,positive group,enteric and gastric excipients group,enteric and gastric pellets group,raw material group,and blank group（No modeling）.The liver tissue was stained with HE and Masson,and the pathological changes of liver fibrosis in each group were observed under a microscope.Measurement of liver function indicators in serum of rats in each group through the microplate test:Alanine aminotransferase（ALT）,Aspartate aminotransferase（AST）,ELISA method was used to determine serum on liver fibrosis indicators:Hyaluronic acid（HA）,Laminin（LN）,Procollagen-Ⅲ（PC-Ⅲ）,Type IV collagen（COL-Ⅳ）.Alkaline hydrolysis method was used to determine the hydroxyproline（HYP）in liver tissue,evaluating the intervention effect of the drug on liver fibrosis rats.Results1.The influence of each factor on the response value is obtained through response surface design:drug-excipients ratio（A）>ethanol concentration（C）>excipients ratio（B）;the optimal process is:excipients:active site=2.84:1;MCC:Starch=1.37:1;ethanol concentration 77.4%.This process was verified,and the molding rate was60.48%,the moisture-absorption rate was 5.84%,and the overall rating was 0.6835.2.The coating pan method is used to prepare the enteric pellets and gastric pellets.The fluidity,roundness and moisture content of the pellets are in compliance with the regulations.The release of three batches of enteric pellets in 0.1 mol/L hydrochloric acid was less than 10%,and the release in p H 6.80 PBS was more than 80%.The similarity factors f₂among three batches of release were 96,92,and 88,all of which were greater than 50,indicating that the releases were similar between batches.Targeted evaluation in vivo showed that entericpellets could smoothly pass through the upper digestive tract to the small intestine to release drugs.3.The results of liver histopathology showed that compared with the model group,the fibrous connection around the duct was weakened,and the degree of liver fibrosis was significantly reduced.The results of index measurement showed that,compared with the model group,the levels of various detection indicators in the positive group,the raw material group,the gastric drug group,and the enteric drug group were reduced,which had statistical significance（P<0.05,P<0.01）.Compared with the gastric drug group,the level of AST in the positive group was higher（P<0.05）,and the other indexes were not statistically significant.Compared with the gastric drug group,the levels of AST,ALT,HA,PC-â…¢,and HYP in the gastric drug group were lower than those of the enteric drug group,and the levels of LN,COL-â…£were higher than those of the enteric drug group,there was no statistical difference（P>0.05）.There was no significant difference between the gastric drug group and the raw material group.Conclusion1.Optimizing the process of preparing anti-hepatic fibrosis active site anti-moisture granules of Periplaneta americana were obtained based on the normalized value of molding rate and moisture-absorption rate as the evaluation index;The active site pellets of Periplaneta americana were prepared successfully by the coating pan method,In vitro and in vivo tests have shown that enteric pellets are not destroyed in the stomach and the drug was releaseed in the intestine.2.The drug group can alleviate the pathological changes of liver in rat liver fibrosis model induced by carbon tetrachloride and reduce various fibrosis indexes.There were no significant differences in the effects of different absorption pathways of Periplaneta americana on liver fibrosis in rats.Therefore,the active site of Periplaneta americana can be prepared into a related gastric drug,which is simpler than the preparation method of the enteric drug,and the coating step is omitted,which can not only reduce the economic cost of the preparation production,but also not reduce the drug’s effect on intervention of liver fibrosis.Through the above studies,experimental basis is provided for the development of new anti-liver fibrosis drugs of Periplaneta americana.