| Phloroglucinol is a pure smooth muscle spasmolytic agent of non-atropine and non-atropine,which can act directly on the smooth muscle of gastrointestinaltract and genitourinary tract.Compared with the commonly used clinical atropine and anisodamine antispasmodic drugs,phloroglucinol has the characteristics of rapid antispasmodic and analgesic effect,remarkable effect and good tolerance.Phloroglucinol has extremely high specificity.Phloroglucinol has been widely used in gynecology,urology and liver enterology.The Spasfon-Lyoc injection was first approved for marketing in France in 1993.The production company is France Spanton.At present,there are oral freeze-dried tablets,injections,freeze-dried powder injections and anorectal suppositories in foreign market.According to the inquiry on the website of CFDA,only domestic injections of phloroglucinol are available.In 2005,five companies had obtained production approvals;The are 48 companies currently registered formulations which are injections,orally disintegrating tablets and oral freeze-drying tablets.There is no application for the listing of phloroglucinol suppositories in China.This subject refers to France's Spafon suppositories as a reference preparation to study the formulation and is intended to prepare phloroglucinol suppositories with the same efficacy as the original suppositories.and compares its quality with the in vivo pharmacokinetics of the animal which providing domestic research and development for the suppository formulation of the variety.Based on laboratory data,According to the new chemical registration category,it is classified into 3 categories,imitating foreign dosage forms.Objective The purpose of this study is to study the screening process for preparation of phloroglucinol suppositories in the laboratory,It establishs effective and controllable quality research methods for phloroglucinol suppositories,and investigates the stability of suppositories under different conditions,and conducts a preliminary pharmacokinetic study in rabbits for phloroglucinol.It provides laboratory data for research and development of phloroglucinol suppositories.Methods1.Prescription process research The hot melt method was used to prepare phloroglucinol suppositories.The prescription composition of self-made phloroglucinol suppositories was determined according to the prescription composition of the original phloroglucinol suppository.The type and amount of surfactants and the time of ultrasonic agitation were selected by single factor test.The star point-response surface method was used to determine the prescription process of phloroglucinol suppository.It is optimized for the formulation process of the trisphenol suppository,It is determined the best prescription process and verified the best prescription.2.Quality research Taking the original preparation agent as a reference,according to the“Quality Testing of Suppositories of the Fourth Edition of the Chinese version of the 2015 edition of the Chinese Pharmacopoeia”?0107?,the appearance,weight difference,fusion time limit,and content of self-made phloroglucinol suppositories were studied.The dissolution rate has also been compared in more detail.3.Stability of the study The appearance,content,related substances,melt time,release,were used as the evaluation index,The influence of temperature,humidity,and light on the stability of phloroglucinol suppositories was investigated through the influencing factors.4.Pharmacokinetics study A LC-MS/MS method was established to determine the plasma concentrations of phloroglucinol in rabbits and a methodological survey was conducted.The pharmacokinetic characteristics of rabbits in vivo were investigated after rectal administration of phloroglucinol suppositories to healthy rabbits.Through DAS 3.0software,the pharmacokinetic data of self-made phloroglucinol suppositories and original pyrogallol suppositories were compared.Results1.Prescription process research The best prescription of the suppository prescription is:the main drug phloroglucin ol116.7mg,trimethylphloroglucinol 116.7mg,the matrix is a fat-soluble matrix?A?,?SDS?0.55%;the best preparation process is as follows:the matrix in the beaker of which the prescription amount of phloroglucinol,trimethylphloroglucinol,SDS is used,and the prescription quantity is called the beaker is heated and melted in a 60?water bath until the matrix is melted and then added to the SDS and continuously stirred until it is dissolved.Phloroglucinol and trimethylphloroglucinol were added to the ultrasonic instrument for ultrasonic agitation for 35 minutes.After stirring,it should be cooled to room temperature and then filled in a bolt coated with lubricant?soap:glycerin:alcohol 1:1:5?.Put the plug in the refrigerator at-2?for 15 minutes.Take out the demoulding,that is.The phloroglucinol suppository prepared by the prescription has smooth appearance,no holes,no cracks,uniform color distribution,uniform distribution and moderate hardness.2.Quality research This product is a milky white torpedo suppository with smooth appearance,no pores,no cracks,uniform color distribution,uniform distribution of the drug,moderate hardness.The results of the three batches of suppository weights prepared in the laboratory showed that the average weights for 20170213,20170807,and 20170614 were 1.6552 g,1.6590 g,and 1.6558 g,respectively.The weight difference limits were 1.5311 to1.7793 g,1.5346 to 1.7834 g,and 1.5316 to 1.7799 g,respectively;The weight of each suppository of the sample was within the limit of the weight difference of each batch of suppository,which was in line with the requirements of the 2015 edition of the Chinese Pharmacopoeia.The time of fusion?the results showed that 3 batches of suppositories all softened within 20 minutes?,and the results were in line with the 2015 edition of the Chinese Pharmacopoeia.Dissolution was measured using a paddle method.Phosphate buffer p H 7.4containing 0.05%Tween-80 was used as the dissolution medium.The stirring paddle speed was 100 r/min.Dissolution measurement results showed that the self-made phloroglucinol suppository and the original pyrogallol suppository had a f2factor of 53.70 for the phloroglucinol dissolution curve and a trimethylphloroglucinol dissolution curve for the f2 factor of 67.11.The dissolution behaviors of the two are relatively consistent.The content of phloroglucinol was determined by HPLC,and the chromatographic column was Nuanalytical Cis?250 mm x 4.6 mm,5?m?.The mobile phase was acetonitrile and potassium dihydrogen phosphate solution?pH 3.0?,gradient elution,detection wavelength 265 nm,column temperature of 30?.Phloroglucinol and trimeth ylphloroglucinol tR were 4.1 min and 9.3 min;phloroglucinol has a good linear relation ship with the peak area in the range of 6.0240.16?g.m L-1concentration,trimethylphloroglucinol has a good linear relationship with the peak area in the range of 6.0540.32?g.m L-1concentration.The linear equations are y=45.919x+0.080?R2=0.9999?,y=49.424x-0.291?R2=0.9997?.The average yield was 99.97%?RSD=0.67%,n=9?,100.34%?RSD=1.2%,n=9?.The average content of phloroglucinol in the three batches of samples?20160701,20160702,20160703?was100.3%,the RSD was 0.61%,the average content of trimethylphloroglucinol was99.96%,and the RSD was 0.93%.3.Stability study The selfmade phloroglucinol suppository and the original research phloroglucinol suppository were investigated for stability investigation.Influencing factors test results are shown below.high temperature:The appearance of selfmade suppository and the original research were melted and deformed,and the content has no obvious change.High humidity:the color of theself-made suppository and the original research have changed deeply,the content of the main drug was reduced,and the dissolution rate of the main drug was slowed down.Light:the color of the selfmade suppository and the original research have changed deeply,the content of the main drug was reduced,and the dissolution rate of the main drug was slowed down.It is indicated that the storage conditions of phloroglucinol suppository need to choose the appropriate temperature and humidity,and avoid the light.4.Pharmacokinetic study In pharmacokinetic study established a method for the determination of phloroglucinol in rabbits,LC-MS/MS liquid phase and mass spectrometry conditions,and PG mass charge ratio:m/z 125-m/z 57;the linear range of PG plasma sample was 5.00-2500.00 ng.m L-1;endogenous substances did not interfere the determination of PG;the precision is good,the recovery rate of the method is good,and the recovery rate meets the requirements;the matrix effect is negligible.The pharmacokinetic parameters of the selfmade phloroglucinol suppository were as follows:AUC?0-t?:1519.36±186.55?g/L*h,Cmax:2657.65±222.91?g/L,tmax:0.25 h,t1/2:3.99±0.15 h.The pharmacokinetic parameters of Spasfon suppository were as follows,AUC?0-t?:1758.79±357.55?g/L*h,Cmax:2392.42±291.91?g/L,tmax:0.75 h,t1/2:1.54±0.72 h.The test of AUC?0-t?and Cmaxax showed that there was no significant difference in AUC?0-t?and Cmaxax between the two preparations.P>0.05.It shows that the self-prepared suppositories are basically the same as the original suppositories,and the bioavailability is equivalent.The tmaxax rank sum test results showed that P<0.05,the difference in tmaxax between the two preparations was statistically significant.The tmaxax of the homemade suppository was less than the tmaxax of the original suppository.It shows that the self-made suppository has a faster onset time.Conclusions1.The phloroglucinol suppositories prepared in this study which have smooth appearance,uniform cut surface,no obvious pores inside,moderate hardness,and the weight difference and melting time limit conform to pharmacopoeia requirements.In vitro drug dissolution was consistent with the original study.2.The RP-HPLC determination method was established and the methodological research results meet the measurement requirements.The average amount of phloroglucinol in the three batches of samples prepared in the laboratory was100.30±0.61%,and the average content of trimethylphloroglucinol was 99.96±0.93%.3.The stability test showed that the phloroglucinol suppository was sensitive to humidity and unstable under high humidity conditions,and it should be stored under dry conditions;it is sensitive to high temperature and need low temperature preservation,it is sensitive to light and need to be protected from light.4.Pharmacokinetic experiments showed that the self-prepared suppository and the original suppository are bioequivalent in vivo,the self-prepared suppository could quickly take effect in rabbits,and the onset time of the self-prepared suppository was faster than that of the original phloroglucinol suppository. |
PDF Full Text Request