The Preclinical Pharmacy Research Of SEM Tablets Based On Network Pharmacology And A Classical Prescription (HH)

Objective:According to the research ideas of the components of traditional Chinese medicine,the effective components were screened from the classic prescription Huanghuai（HH）compound by using network pharmacology technology,and preclinical pharmaceutical research was carried out on them.Methods:1.In the research of network pharmacology,TCMSP,Uni Prot,Gene Cards,STRING and DAVID databases were used to construct the"component-target-pathway"network of HH for the treatment of diseases through Cytoscape 3.7.2 software.The topological parameters were obtained by network analysis,and the key targets were selected.The key components in the network and the key targets were theoretically verified by molecular docking;the content of each key component in the prescription was determined by HPLC,and the new compositions were combined according to the content proportion of each component（referred to as FEM and SEM respectively）2.Use aborted uterus test,capillary method and tail-docking method,combined with changes in biochemical index levels to study its hemostatic activity.The anti-inflammatory activity was studied by the experiment of egg white-induced plantar swelling in mice.The pharmacological activity of HH,FEM and SEM was compared.3.According to the physical parameters（Forming rate,hygroscopicity,angle of repose,Karl coefficient）of oral solid preparation of traditional Chinese medicine,the central composite design response surface method was used to optimize the preparation process and parameters of SEM tablets intermediates;in the research of forming process of SEM tablets,the method of weight coefficient weighted comprehensive score was used to select the types and dosages of lubricants and disintegrats.4.The physical fingerprints and UV-Vis fingerprints were used to establish the control chromatograms;the HPLC“quantitative analysis of multi-components by single-marker”method was used to determine the content of the related components,and the NIR quantitative simplification model was established to predict the content of the related components in the intermediates of SEM tablets.5.The related components in SEM tablets were qualitatively identified by thin layer chromatography;the hardness,weight difference,brittleness and disintegration time limit of the tablets were specified according to the method specified in the fourth part of Chinese Pharmacopoeia（2020 Edition）;the dissolution and release of the tablets were determined by paddle method（general principles of Chinese Pharmacopoeia 2020 Edition）According to the general rule 0512 of Chinese Pharmacopoeia（2020 Edition）,the content of related components in SEM tablets was determined.6.According to the guiding principles of drug stability experiment in Chinese Pharmacopoeia（2020 Edition）,the influencing factors test of SEM tablets were carried out,including high temperature,high humidity and strong light irradiation test,in order to provide the basis for the determination of production,packaging,storage and transportation conditions of SEM tablets.Results:1.Using network pharmacology to screen out the key components of HH for the treatment of abnormal uterine bleeding were quercetin,wogonol,kaempferol,baicalein,acacetin andβ-sitosterol.The key targets were PTGS2,AKT1.The molecular docking results of TP53 and TNF show that the docking binding energy of key components and key targets was less than-5.0 k J·mol^-1,indicating that they had strong binding activity.The quercetin,wogonin,kaempferol,kaempferol baicalein,acacetin andβ-sitosterol may bind to key targets stably and play a significant role.2.According to the content determination results,it was finally determined that kaempferol,baicalein,wogonin,quercetin and acacetin were combined into composition 1（hereinafter referred to as FEM）in the ratio of4.15:0.45:0.13:0.11:0.02.Baicalin,kaempferol,baicalein,wogonin,quercetin and acacetin were combined into composition 2（hereinafter referred to as SEM）in the ratio of 9.23:4.15:0.45:0.13:0.11:0.02.3.The validation results of the drug efficacy test showed that:（1）Hemostatic activity:HH,FEM and SEM all have very significant hemostatic activity,and compared with the original compound HH,there is no significant difference in efficacy between FEM and SEM and HH;（2）Anti-inflammatory activity:HH and SEM had significant anti-inflammatory effects,and SEM had more significant anti-inflammatory effects than others.4.Preparation process of SEM tablet intermediate:Weigh SEM raw material powder and lactose(m _SEM:m_lactose=5:4),mix for 15 minutes;add 70%Ethanol,make soft material,granulate through a 14-mesh sieve,dry at 60℃,and granulate,ready to be obtained.5.Take an appropriate amount of the SEM tablet intermediate,add 1%croscarmellose sodium and 2.5%talc,well mixed,and press the tablet to obtain the SEM tablet.6.The quality standard of SEM tablet intermediate:physical fingerprint spectrum（similar degree 0.847～0.989）,ultraviolet fingerprint spectrum（similarity 0.926～0.973）,the relative standard deviation calculated by HPLC of quantitative analysis of multi-components by single-marker was less than5%,and there was no significant difference,it can be used for the quality evaluation research of multi-component intermediates.And the established NIR quantitative determination model was accurate.7.SEM tablet quality standard:prescription（baicalin 176.69 g,kaempferol 79.44 g,baicalein 8.61 g,wogonin 2.49 g,quercetin 2.11 g,and acacetin 0.04 g）;traits（light yellow-green tablets,Slight gas,slightly bitter taste）;inspection items:appearance,tablet weight difference,friability,disintegration time limit,hardness,etc.all meet the requirements;content determination（the content of baicalin in each tablet was not less than 137.39mg,baicalein The content was not less than 9.70 mg,and the content of kaempferol was not less than 66.64 mg）;dissolution rate（At 30 min,the dissolution limit of baicalin and kaempferol is 80%of the labeled amount）.8.The results show that the SEM film is stable under strong light,sensitive under high temperature and high humidity,and should be stored at room temperature and sealed.Conclusions:According to the research ideas of multi-component chinese medicine,this paper had completed part of the preclinical pharmacy research work of SEM tablets,and carried out the screening of the active ingredient group of Huanghuai compound for the treatment of abnormal uterine bleeding,and verified it,and combined it into a new one according to the principle of"component Chinese medicine"The composition（SEM）was developed into a tablet.This article studied the process and quality standards of its intermediates and finished products,and its stability was good.