| In recent years,clinical infection is becoming more and more prominent,and bacteria and fungal have become increasingly resistant,so it is necessary to develop new drugs with strong antibacterial activity,broad antibacterial spectrum,and good resistance.Dryopteris fragrans?L.?Schott,a member of the Dryopteris genus?Dryopteris Adans.?,has been used for a variety of dermatological diseases such as psoriasis,acne,rash and dermatitis.Pharmacological studies have shown that Dryopteris fragrans has various activities such as antibacterial,antipruritic and anti-allergic activities.More than ten phloroglucins and their derivatives were isolated from Dryopteris fragrans in the early stage of our research group.Through the screening of antibacterial spectrum,it was found that flavaspidic acid BB had significant antibacterial activity compared with the commonly used clinical external drugs,and had a better inhibition on the formation of biofilm,one of the main drug-resistant factors of bacteria It is planned to develop it into a new bacterial biofilm inhibitor anti-infection Class I new drug,which is supported by a major special project of Guangdong Provincial Department of science and technology.The purpose of this study is to complete the pharmaceutical research of the new drug according to the requirements of the Class I new drug application materials,including the research on the quality standard of the flavaspidic acid BB.The preparation process of flavaspidic acid BB cream was determined and its quality standard was established;in order to select the suitable enhancer for the in vitro transdermal absorption,the in vitro transdermal characteristics of flavaspidic acid BB cream were studied.At the same time,record the original data truthfully,carefully and normatively for on-site audit,so as to lay a foundation for the application of new drugs.This part of the study is as follows:1.Study on the quality standard of flavaspidic acid BB:Refer to the relevant provisions of Chinese Pharmacopoeia?2015 Edition?,the properties,physical and chemical identification,loss on drying,residue on ignition,heavy metals and other items were examined,and the relevant substances and contents were determined by ultra high performance liquid chromatography?UPLC?.The influence of external conditions such as high temperature,high humidity and strong light irradiation on the samples was investigated by the experiment of influencing factors.2.Study on the preparation of flavaspidic acid BB cream:By screening the matrix formulation for the preparation process,the method of adding the oil phase and the water phase,the orthogonal test is used to determine the amount of the oil phase and the emulsifier to optimize the optimal process of cream preparation.3.Study on the quality standard of flavaspidic acid BB cream:the indicators such as character,identification,particle size and microbial limit were investigated,the relevant substances and contents were determined by UPLC method,At the same time,accelerated stability and long-term stability were investigated according to the guiding principle of 9001,Part IV,Chinese Pharmacopoeia,2015 edition.4.Study on transdermal absorption of flavaspidic acid BB cream in vitro:Modified Franzdiffusion cell was adopted using abdominal skin of isolated male rat as transdermal barrier.The content of flavaspidic acid BB was determined by UPLC.The accumulative transdermal amount(Q24 h)and percutaneous permeability(Jss)within 24 h were calculated,and compared with Flavaspidic acid BB cream without transdermal enhancer,and calculated enhancement ratio?ER?.The results were shown as followed:1.Study on the quality standard of flavaspidic acid BB:flavaspidic acid BB was yellow powder,odorless,slightly hygroscopic,soluble in ethyl acetate and acetone,almost insoluble in water,melting point was 165??170?,adding ferric chloride test solution showed reddish-brown,with maximum absorption at a wavelength of 296 nm.By UPLC method,the content of impurity I in three batches of raw materials was 0.3294%?0.3485%,and the content of main component was 97.90%?98.36%.Under the conditions of high temperature,high humidity and strong light irradiation,the contents of the 5th day?97.51%,97.56%,97.40%?and the10th day?97.44%,97.24%,97.17?of the sample?No.:GC-CZ-180906?were close to 0 day.2.Study on the preparation of flavaspidic acid BB cream:Based on the matrix prescription and orthogonal factors,the optimal dosage of the matrix was 6 g vaseline,5 g stearyl alcohol,4 g peregal O-9 and 12 g glycerol.With peregal O-9 as the emulsifier and azone as the transdermal absorption promoter,the drug was dissolved in the oil phase,the emulsifying temperature was 65?.The best preparation process was obtained by slowly pouring oil phase into water phase.3.Study on the quality standard of the flavaspidic acid BB cream:the product was light yellow cream,the particle size was less than 180?m,Staphylococcus aureus,P.Aeruginosa and other molds were not detected.After the investigation of UPLC methodology,results showed that the content of impurity I in the three batches of cream was 0.3294%?0.3485%,and the content of flavaspidic acid BB was 97.05%?99.34%.Through the investigation of accelerated stability for 6 months and long-term stability for 12 months,the appearance of three batches of cream was light yellow,the particle size was less than 180?m,the impurity I and the percentage content fluctuation of main component labeling amount were all within the scope of formulation,which was in line with the provisions of the quality standard.Therefore,the validity period of this product is tentatively 12months.4.Study on transdermal absorption of flavaspidic acid BB cream in vitro:with 35%ethanol-normal saline solution as the receiving solution,The results showed that the Q24h of flavaspidic acid BB cream containing azone?1%,2%,3%,4%?,1%menthol,1%propylene glycol,1%oleic acid,1%azone+1%menthol,1%azone+1%propylene glycol,1%azone+1%oleic acid,1%menthol+1%propylene glycol and other 11 penetration enhancers were?82.96±7.15?,?80.17±0.66?,?78.22±1.87?,?73.53±1.24?,?35.65±2.23?,?34.02±1.73?,?42.68±2.66?,?33.94±1.37?,?34.16±1.54?,?46.78±1.21?,?43.66±1.69??g/cm2,respectively.Jss value were?5.26±0.10?,?4.69±0.12?,?4.45±0.45?,?4.00±0.06?,?3.74±0.33?,?3.23±0.18?,?3.73±0.53?,?3.14±0.47?,?3.54±0.11?,?3.98±0.34?,?4.34±0.14??g/cm2·h,respectively.ER were 2.055,1.831,1.738,1.564,1.462,1.263,1.456,1.227,1.385,1.557,1.698,respectively.The permeation test results of three batches of flavaspidic acid BB cream were 5.257±0.10,5.053±0.13,and 5.585±0.07.Conclusion:?1?According to the requirements of Class I new drug application,this study established the quality standard of the crude drug of flavaspidic acid BB.?2?The preparation process of flavaspidic acid BB cream was determined by screening the prescription matrix and optimizing the preparation process;at the same time,the quality standard of the cream was formulated.?3?Through the accelerated stability and long-term stability inspection,it was determined that the product was sealed and preserved in the shade,and the validity period was tentatively 12 months.?4?Through the study of transdermal characteristics in vitro,1%azone was used as the transdermal absorption enhancer of flavaspidic acid BB cream,and the cream can effectively produce local effects.The study of preclinical pharmacy was completed,which laid a foundation for the application of Class I new drugs. |
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