Study On Preparation Of Chongqi Colon Targetting Pellet Capsules And The Interfering On UC Model Induced By DSS In Mice

Objective:To study the compound preparation of Chinese medicine for ulcerative colitis?UC?"Chongqi Colon Targetting Pellet Capsule?PA-CTPC?",and to investigate its preparation technology,quality standard,stability and preliminary pharmacodynamics.Methods:1.With the extraction rate of active ingredients polyphyllin ?and ? as index,the ethanol extraction process of Paridis Rhizoma was optimized by L₉?3⁴?orthogonal test,the water sedimentation and drying process of alcohol extract of Paridis Rhizoma was optimized by single factor test,and the extraction rate of total polysaccharides of Astragali Radix as index,the water extraction process of Astragali Radix was optimized by L₉?3⁴?orthogonal test,and the single factor test was used to optimize the extraction process of Astragali Radix Ethanol precipitation and drying process of water extract;taking roundness and yield of pellets as evaluation indexes,the types and dosage of excipients and concentration of wetting agent were investigated by single factor method,and extrusion-spheronization process of pellets was optimized by Box-Behnken response surface method;pellet coating formulation was optimized by single factor method;Cumulative release of polyphyllin ?and ? was evaluated as index,release properties of pellets in vitro.2.The content of the main effective components of polyphyllin ?and ? in the preparation was determined by high performance liquid chromatography?HPLC?.The content of total polysaccharides of Astragali Radix in the preparation was determined by phenol-concentrated sulfuric acid method.The draft quality standard of PA-CTPC was formulated,and the preliminary stability test of the preparation was carried out.3.The UC model of mice was induced by 4%DSS solution drinking freely.After successful modeling,UC mice were treated with corresponding doses of saline and PA-CTPC respectively.Colorectal tissues were taken after the mice were executed.The intervention effect of PA-CTPC on UC was investigated by disease activity index,pathological morphology and the degree of intestinal mucosal damage.Results:1.The optimum preparation process of total saponins of polyphyllin was as follows:extracting Paridis Rhizoma pieces,crushing them into the crudest powder,refluxing with 6 times 80%ethanol for 3 times,1.5 hours each time;combining ethanol extract,filtrate decompression,recovering ethanol and concentrating it to relative density of about 1.10?60??,adjusting pH to 8.0⁹.0 with Ca?OH?₂ saturated water solution,centrifuging,extracting supernatant,and adding 4 times water for refrigeration.The precipitation was dried by vacuum drying?65⁷0?,-0.08MPa?and then crushed.The best preparation process of Astragali Radix is:Astragali Radix pieces,add 8 times amount of water to reflux 3 times,1.5h each time,merge the extract,filtrate,filtrate decompress and concentrate to relative density of about 1.10?60??,centrifugation,take supernatant,add alcohol to make alcohol content 70%,static cool storage?2¹0??24h,filtration,precipitation with appropriate water mixing,spray drying,crushing.The optimum formulation of pellets as follows:the ratio of microcrystalline cellulose to powder is about1:1,85%ethanol as binder,extrusion rate is 50 r/min,rolling speed is 1150 r/min,and rolling time is 4 min.The yield of pellets is 83.67%,roundness is 17.75°.The optimum formulation of pellets coating is:Eudragit S100 is coating solution,coating weight gain is15%,plasticizer TEC dosage is 10%,anti-adhesive dosage of glyceryl monostearate is 2%.The coated pellets have good colonic targeting drug release characteristics when the dosage is 2%.The accelerated stability test of PA-CTPC for 6 months and the long-term stability test for 12 months showed good quality stability.2.According to the determination results of the main active ingredients by HPLC,the sum of the contents of polyphyllin ?and ? in the prescription should not be less than 10mg/g,and the content of total polysaccharides of Astragali Radix should not be less than60 mg/g,according to the determination results of the main components by ultraviolet spectrophotometry.3.Compared with the blank group,the DAI score of the model group was significantly higher?P<0.01?,and the damage of colon tissue was obvious?P<0.01?.Under the optical microscope,it was found that the mucosal epithelium and intestinal gland of the model group were severely damaged,a large number of lymphocytes infiltrated into the submucosa?P<0.01?,and the content of D-LA and DAO increased significantly?P<0.01?.Compared with the model group,the DAI scores of PA-CTPC in middle and high dose groups were significantly lower?P<0.05?.The naked eye observation showed that only a few ulcers existed in colon mucosa of mice in middle and high dose groups of PA-CTPC,and no edema,congestion and granular erosion were found.The glands of mucosa were arranged neatly under optical microscope,and only a few inflammatory cells infiltrated,which was not significant compared with the blank group.The difference was significant?P>0.05?.Compared with the model group,the pathological score of colon tissue was significantly lower?P<0.05?,the contents of D-LA and DAO in serum were significantly decreased?P<0.05?,and the content of DAO in serum of mice in high dose PA-CTPC group was significantly decreased?P<0.01?.There was no significant difference in DAI score,colon histopathological score,D-LA and DAO content between PA-CTPC low dose group and model group?P>0.05?.Conclusion:The preparation process of PA-CTPC is simple and effective,the quality standard control method is accurate and reliable,and the preparation is stable.Preliminary pharmacodynamic studies showed that PA-CTPC could improve the survival status of UC mice induced by 4%DSS to a certain extent,delay and inhibit the development of UC in mice.