Process Influence In Vivo Of Reduning Injection Co-administration

Reduning injection(RDN)is one of TCM injections for treatment of upper respiratory tract infection from which is composed of,Lonicerae japonicae Flos(Lonicera japonica Thunb.),Artemisiae Annuae Herba(Artemisia annua L.)and Gardeniae Fructus(Gardenia jasminoides Ellis.).RDN is always often combined with cefuroxime sodium(CNa)in the treatment of pneumonia,mycoplasma pneumonia and bronchitis in children.Moreover,it always co-administrates with ribavirin(RBV)to treat hand,foot and mouth disease in children.Although these combinations are common,there were few studies on the interactions in vivo.In this study,we studied the drug influence of RDN and CNa,RDN and RBV in vivo process.Literature researchA synthetical review was given to the present study of drug interactions in pharmacokinetics,in order to provide scientific reference for the study.Experimental study1.Study on the process influence in vivo between RDN and CNa1.1 Identification of the metabolites of Reduning injection in rat plasma,bile,urine and feces after intravenous administrationAs a result,we determined 14 metabolites of geniposide,including oxidation,dehydration,hydroxymethylene loss,hydrolysis,ring-opened,cysteine conjugation and glucuronidation conjugation of aglycone.9 metabolites of geniposidic acid,consisting of dehydration,ring-opened,double-bond reduction and cysteine conjugation.6 metabolites of secoxylogain including hydrolysis,hydroxymethylene loss,hydroxylation and ethylation.12 metabolites of chlorogenic acid,containing decarboxylation,hydrolysis,methylation,acetylation,cysteinylglycine conjugation and glutathione conjugation.1.2 Pharmacokinetic influence of RDN co-administration with CNaIn order to study the effects of RDN combined with CNa,an UHPLC-LTQ-Orbitrap-MS method had been establised to determine the four kinds of iridoid glycosides,3 kinds of organic acids and cefuroxime.Meanwhile,a semi-quantitative method method of 12 metabolites based on the preliminary research.After single co-administration,the AUC(0-t)and Cmax values of iridoid glycoside decreased.The Cmax of ring-opened metabolite of geniposide increased,and the area under the curve increased significantly.The peak concentration of the cysteine conj ugation product and the oxidation product of geniposide was slightly increased.The relative content of the hydroxylated ethylated conjugation product and the hydroxymethylene loss product of the secoxyganin were reduced.The AUC(0-t)and Cmax values of organic acids.The relative content of the chlorogenic acid methylation product and the acetylated product were decreased.The AUC(0-t)of cefuroxime decreased,and there was no significant difference in T1/2 and Cmax.After multiple combinations,the AUC(0-t)and Cmax values of shanzhiside,genipin-1-?-gentiobiose and secoxyganin decreased significantly.The relative content of the metabolites of geniposide and secoxyganin decreased,and the glucuronidation product was the most significant.The parameters AUC(0-t)and Cmax decreased,T1/2 had no significant changes.The relative content of chlorogenic acid methylation product and acetylation product in plasma decreased.There was no significant changes of cefuroxime1.3 Distribution influence of RDN co-administration with CNaA determination method of above eight ingredients had been set up to explore the effects on tissue distribution after combination.After intravenous administration of RDN,the ingredients distributed rapidly in main organs and mainly accumulated in the liver and kidney.At 10 min,the tissues were most widely distributed and subsequently decreased.The cefuroxime mainly accumulated in the liver and kidney.After co-administration,the contents of the iridoid glycosides,organic acids and cefuroxime in main tissues decreased.The peak of distribution delayed.1.4 Metabolism influence of RDN co-administration with CNaThe real-time quantitative polymerase chain reaction(Q-PCR)was applied to determine the mRNA expression levels of CYP450.RDN can induce the mRNA expression of CYP1A2,CYP2C9 and CYP2C19,inhibite CYP2A6.The effect of cefuroxime on these seven metabolic enzymes was not obvious.At the mRNA level,RDN combined with CNa significantly induced the activity of CYP2B6,CYP2C9,CYP2C19 and CYP2E1,had an inhibition on CYP2A6.1.5 Excretion influence of RDN co-administration with CNaA determination method of above eight ingredients had been estabolished to explore the effects on excretion after combination.Intravenous administrating respectively,the rats excreted RDN mainly by urine and bile,excreted CNa by urine.All components urine excretion wrere concentrated at 0-4 h,bile concentrated at 0-2 h.After combination,the contents of shanzhiside,genipin-1-?-gentiobiose and cefuroxime in urine increased.All the compounds excreted less in bile.2.Study on the process influence in vivo between RDN and RBV2.1 Pharmacokinetic influence of RDN co-administration with RBVAfter single co-administration,the AUC(0-t)and Cmax values of iridoid glycosides and organic acids were significantly reduced.Moreover,the Cnmax of cysteine conjugation product of geniposide and methylation product of chlorogenic acid,the peak of decreased.The Tmax of hydroxymethylene loss product of geniposide,methylation product of chlorogenic acid.The AUC(0-t).Cmax and T1/2 values of RBV were obviously increased.After multiple combinations,the Cmax values of iridoid glycosides and the organic acids significantly reduced.The relative contents of cysteine conjugation product,oxidation product of geniposide;hydroxylation and ethylation product,hydroxymethylene loss product,aglycone hydrolysis product of secoxyganin;methylation product of chlorogenic acid decreased.Meanwhile,the Tmax of cysteine conjugation product,dehydration product and hydroxymethylene loss product of geniposide;hydroxylation and ethylation product,aglycone hydrolysis product of secoxyganin were delayed.The AUC(0-t),Cmax and T1/2 values of RBV were significant increased.2.2 Distribution influence of RDN co-administration with RBVIn the ribavirin injection,the order of distribution was liver>spleen>kidney>lung>heart,and the peak time of liver was delayed compared with other organs.After combination,the contents of iridoid glycosides increased in the heart and those of organic acids in the kidney increased.Ribavirin mainly distributed in liver and the Tmax was more delayed than the single administration.2.3 Metabolism influence of RDN co-administration with RBVRibavirin had little effects on liver drug enzymes.After co-administration,the mRNA expressions of CYP1A2,CYP2A6 and CYP2B6 in rats were inhibited.2.4 Excretion influence of RDN co-administration with CNaRibavirin had a higher cumulative excretion rate in urine than bile when administration alone.After intravenous co-administration,the contents of geniposide in urine decreased.All the compounds of RDN excreted less in bile.Comparing to administration alone,the content of ribavirin reduced in both urine and bile.