| Azatadine maleate, an antihistamine developed in1971, is a potentlong-acting H1receptor antagonist. Azatadine maleate has the similarpharmacology such as anti-serotonin, anticholinergic and antianaphylacticactivity as cyproheptadine, but less sedation effect. The mechanism related tocompetitively inhibition of H1-receptors, anti-vasodilatation and reducing thepenetration of capillary and symptom of edema. The azatadine maleat tabletprouduced by Schering Canada Incorporation was first approved by FDA in2002, the brand name are Opitimine which containing1mg maleateazatadine, and the other one containing maleat azatadine1mg andpseudoephedrine sulfate120mg was named Trinalin. The preparation ofmaleat azatadine still blong to blank in Chinese.Aim: To date, there are few reports on the pharmacokinetics study ofazatadine maleat in human, this study aimed to develop a high sensitiveLC-MS/MS method to determine the concentration of azatadine in humanplasma and apply the method to investigate the pharmacokinetics propertyafter oral azatadine maleat tablets following single and multi-dose. The studyfirstly developed and applied the LC-MS/MS method to determinate thepharmacokinetics property of azatadine in healthy subjects after single and multi oral dosing in the international. The study will supply necessaryinformation for the pharse â…¡ clinical trails and provide more data forrational useage of this medication in clinical practice.Methods: the study include three partsPart1A sensitive method using liquid chromatography with tandem massspectrometric detection （LC–MS/MS） was developed and validated for theanalysis of antihistamine drug azatadine in human plasma. Procedures:Analytes were extracted from human plasma by liquid/liquid extraction usingethyl acetate. The organic phase was reduced to dryness under a stream ofnitrogen at40C and the residue was reconstituted with120μL of mobilephase.5μL of the resulting solution was injected onto the LC–MS/MS system.An Agilent ZORBAX TC-C18column was used to perform thechromatographic analysis. The mobile phase consisted of ammonium formatebuffer0.010M （adjusted to pH4.3with1M formic acid）/acetonitrile （20:80,v/v）. The tandem mass spectrometric detection mode was achieved withelectrospray ionization （ESI） iron source and the multiple reaction monitoring（MRM） was operated in positive ion modes. The mass transition was291.3â†'248.2m/z for azatadine,383.3â†'337.3m/z for Loratadine, respectively. Thenebulizer pressure:55psi, fragmentor voltage:115V （azatadine） and100V（IS）, and collision energy:14eV （azatadine） and22eV （IS）.Part2The pharmacokinetics property of azatadien maleat tablets following asingle dose administration in healthy volunteers was stuied.20healthyvolunteers was randomly divided into two groups on gender parity, low doseand high dose groups were given single dose of maleat azatadien tablet with1pill（containing1mg azatadine maleat per pill） and2pills respectively. Bloodsamples （4mL） were collected at0h （pre-dose） and0.5,1.0,2.0,3.0,4.0,5.0, 6.0,8.0,10.0,12.0,24.0,36.0and48.0h （post-dose）. The samples weretransferred to heparinized tube and centrifuged at3000×g for10min. Plasmawas processed and detected the concentration of azatadine by LC-MS/MSmethod. The plasma concentration–time data was analyzed with the help ofWinNonlin6.0. All pharmacokinetics parameters were obtained by thesoftware.Part3The pharmacokinetics study of Multi-dose was following the singledose study of1mg maleat azatadine. Each volunteer was given1pill for twicea day at8:00and20:00for six consecutive days. The test medication wastaken with200mL water. Blood sample was collected on the third to the fifthday before taken the first dose at8:00, on the six day, blood sample wascollected at0h （pre-dose） and0.5,1.0,2.0,3.0,4.0,5.0,6.0,8.0,10.0,12.0,24.0,36.0and48.0h （post-dose）. As the single dose study, all volunteerswere supplied with low-fat food at least4hours after taking the testmedication. The method of sample and data processing are same as the singledose study.Results:1. The method of LC-MS/MS was applied for detecting azatadine in humanplasma after single and multiple dosing. the linear range for azatadien werefrom0.05to4ng·mL-1; The lowest limit of quantification was0.05ng·mL-1;the RSD of intra-precision and intra-precision for three concentrations wereless than11.57%and9.17%repectively; the recovery rates were above70%;other methodology parameters, such as medium effect, stability, are all meetthe criterion of determination of biological samples.2. The pharmacokinetics parameters of low dose study following single doseare shown as follows: t1/2:11.72±2.01h; AUC0-48:11.22±4.05ng·h·mL-1; Cmax:0.795±0.187ng·mL-1. Pharmacokinetics parameters of high dose study:t1/2:10.62±2.06h; AUC（0-48）:18.80±6.98ng·h·mL-1; Cmax:1.444±0.245ng·mL-1. There is no significant different in elimate half-life timebetween the low and high dose. The parameter of AUC（0-48）and Cmaxshows atendency of liner relation with dosing. Comparing the parameters betweenmale and female, the result shows no gender different exsiting after singledose oral administration of azatadine maleat.3The pharmacokinetics study of Multi-dose was following the single dosestudy of1mg azatadine maleat. Each volunteer was given1pill for twice aday at8:00and20:00for six consecutive days. The pharmacokineticsparameters of multi-dose are showon as fllows: t1/2:10.46±2.26h; AUCss:6.89±1.14ng·h·mL-1，AUC（0-48）:13.13±2.49ng·h·mL-1; Cmax:0.87±0.18ng·mL-1. The steady state pharmacokinetics parameters, such as Cmax, t1/2andAUC0-∞, show no significant different compare with the single dose study.The result indicates that there is no accumulation effect after multi-dose inhealthy subjects. To compare the pharmacokinetics parameters of male withfemale, the result shows there is no gender different exsiting after multi-doseoral administration of maleat azatadine.Conclusion:1. A HPLC/MS/MS method was developed for the determination of totalazatadine in human plasma for the first time.The method has been validatedand applied for pharmacokinetics study of azatadien maleat tablet followingoral administration.the method shows rapid, sensitive and specific property indetecting azatadine in human plasma.2. The AUC （0-t） and Cmax of single dose maleat azatadine tablet showstendency of liner relation with the dosing. The results indicate that the azatadine shows linear pharmacokinetics characteristics after oraladministration in healthy subjet.3. The steady state pharamacoketnics parameters show no significant differentcompare with the single dose study. The results indicate that there is noaccumulation effect after multi-dose in healthy subjects. Continuous Oral1mgmaleat azatadien tablet shows a good safety profile.4. There is no gender different exsiting after single and multi-dose oraladministration of maleat azatadine. |
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