| Traditional Chinese medicine injection(TCMI)is a sterile preparation refined from single or compound extracts of traditional Chinese medicine(TCM),which plays an irreplaceable role in the prevention and treatment of clinical diseases since its advantages of high bioavailability and strong practicability.In practice,it is frequently combined with other drugs to achieve multiple therapeutic effects.However,the basic research on the stability,safety and effectiveness of the solution after the compatibility of TCMI remains weak.Introspectively,the pharmacodynamic material basis of TCM is vital to quality control,unfortunately,there still remains unclear,which hinder the safety and quality evaluation of TCMI,quality control of TCM,et al.Therefore,to surmount the above bottlenecks,Kudiezi injection(KDZI)which was a refined water extract prepared from the dried whole herb of Ixeris sonchifolia Hance(Is H)was taken as the research object in this study,investigating the compatibility and stability of the injection based on the pharmacodynamic material basis of TCM.First,the potential core target of Is H in the prevention and treatment of coronary heart disease(CHD)was forecasted by bioinformatics technology.Based on the “affinity mass spectrometry(AMS)-atomic force microscope(AFM)” imaging technology,the active components of Is H were screened and confirmed and the possible mechanism of action was predicted.Secondly,under the guidance of the core target protein and potential signal transduction pathway closely related to the growth of cardiomyocytes,the other potential pharmacodynamic material basis of Is H were confirmed based on the targets.Finally,according to the “stability study and evaluation”instruction in the document “Reevaluation of safety of TCMI”,the compatibility stability of KDZI was investigated.The purpose of this research aims to provide reference for the quality evaluation of TCMI,the material basis research and the quality control of TCM,and the clinical safety and accurate drug use.The experimental contents were as follows:1.The identification of effective substances about the treatment of CHD with Is H via“AMS-AFM” imaging technologyIn this study,firstly,we found that Vascular endothelial growth factor receptor 2(VEGFR2)located on the cell membrane of Vascular endothelial growth factor(VEGF)signal transduction pathway was potential core target protein for the treatment of CHD through network pharmacology and bioinformatics research.Secondly,based on VEGFR2,AMS was applied to screen the active components of Is H,which showed that luteolin-7-O-glucuronide was a potential active molecule.Then,the active molecule changed the molecular morphology of VEGFR2 by AFM imaging and molecular docking simulated the interaction mechanism of luteolin-7-O-glucuronide and VEGFR2.Finally,the activity of H9c2 cells was detected by methyl thiazolyl tetrazolium(MTT)colorimetry and the cell activity of each group was compared by statistical analysis,which was displayed that luteolin-7-O-glucuronide could regulate VEGFR2 to play a balance role in CHD,that was to say,it had a growth protective effect on the injured H9c2 cells.Overall,luteolin-7-O-glucuronide was the pharmacodynamic material basis of Is H.Concurrently,this study provided a possible mechanism for its treatment of CHD.2.Study on other potential pharmacodynamic material basis of Is H based on the targetsIn this module,guided by the VEGFR2 and the target closely related to the growth of cardiomyocytes in the VEGF signal transduction pathway in “Experiment 1”,the effective components were retrieved in the list of 53 main “chemical components-targets” of Is H.The results certificated that adenosine and caftaric acid were all related to proto oncogene tyrosine protein kinase(SRC),at the same time,luteolin-7-O-glucoside,cichoric acid and apigenin-7-O-glucuronide were all contacted to Ras protein.It was speculated that the active components may play a pharmacological role by activating the expression of VEGFR2,the upstream target protein,thus activating the expression of downstream SRC and RAS effector protein.Therefore,the affinity scores and binding sites of the “potential active ingredients-VEGFR2” were confirmed by molecular docking technology,and the interaction mechanism was explained,which revealed a stable hydrogen bond between them.The results indicated that the five components were the potential active substance basis of Is H.3.The compatible stability of KDZI with two kinds of clinical diluents and levocarnitine injection was studiedIn this experiment,we simulated the clinical drug proportion,and mainly investigated the stability of the solution from the aspects of appearance,p H,UV spectrophotometry,insolubleparticles and the content changes of six effective components.Specifically,at 4 ?,room temperature and 30 ?,three batches of KDZI(20181004,20181005 and 20181006)coupled to 5 % glucose injection,0.9 % sodium chloride injection and their compatibility with Levocarnitine injection were investigated.The results indicated that the compatibility solution of the same batch number had clear appearance and no impurity.The p H value,UV spectrophotometry and the content of each effective component changed steadily in 0-8 h at the same temperature.However,when the temperature or injection batch number changed,the p H value and UV Spectrophotometry of the compatible solution would slightly fluctuate but not affected the solution properties,and the content of each active ingredient changed relatively significantly.Additionally,according to the requirements of 2015 edition of?Chinese Pharmacopoeia? on the number of insoluble particles of “intravenous injection with a labeled volume of 100 m L or more”,the insoluble particles of each compatibility solution containing 5 % glucose injection were not in conformity with the specified range,excepted that the compatibility solution of 4 ? 20181006 batch with this injection and levocarnitine injection was in conformity with 0-4 h.However,the compatibility results of0.9 % sodium chloride injection suggested that clinical compatibility should be done at room temperature as much as possible,and should be input with little time.4.Study on the compatibility stability of KDZI with common solvents and vinpocetine injectionThe preparation of the compatibility solution was the same as “Experiment 3”.The results revealed that in the 0-8 h,all the compatibility solutions with the same temperature and batch number,the same temperature and different batch number or different temperature and the same batch number were clear and free of impurities.The p H value and UV Spectrophotometry of the solution changed in a relatively small range,while the content of the effective components changed significantly under the influence of temperature and batch number.However,different from the results of “Experiment 3”,the number of insoluble particles in each compatibility solution of this series did not meet the requirements,so its compatibility stability still needs to be further confirmed. |
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