[生物医药论文精品]盐酸坦洛新及其缓释微丸犬药代动力学研究

1盐酸坦洛新及其缓释微丸犬药代动力学研究摘要目的考察盐酸坦洛新原料药及其缓释微丸在BEAGLE犬体内的药动学行为，评价该缓释制剂的缓释特征。方法BEAGLE犬分别口服给予盐酸坦洛新原料药及缓释微丸制剂后，采用LC/MS/MS法测定犬血浆中各时刻坦洛新的药物浓度，绘制血药浓度时间曲线，并计算药动学参数，对主要药动学参数进行统计学分析。结果BEAGLE犬口服给予盐酸坦洛新原料药及缓释微丸后，血浆中药物达峰时间TMAX、消除半衰期T1/2均存在显著性差异P005；两组峰浓度CMAX经剂量折算后存在显著性差异P005。结论与盐酸坦洛新原料药相比，BEAGLE犬口服缓释微丸后，吸收及消除速度减慢、血药峰浓度降低，符合缓释制剂的药动学行为，判断该制剂具有一定程度的缓释特征。关键词盐酸坦洛新；缓释微丸；LC/MS/MS；药动学ABSTRACTOBJECTIVETOSTUDYTHEPHARMACOKINETICBEHAVIORSOFTAMSULOSINHYDROCHLORIDEBULKDRUGANDITSSUSTAINEDRELEASEPELLETSINBEAGLEDOGSANDEVALUATETHESUSTAINEDRELEASEPERFORMANCEOFTHEPELLETSMETHODSTHEPLASMACONCENTRATIONSOFTAMSULOSINWEREDETERMINEDBYLC/MS/MSMETHODAFTERASINGLEORALADMINISTRATIONOFTAMSULOSINHYDROCHLORIDEBULKDRUGANDITSSUSTAINEDRELEASEPELLETSTOBEAGLEDOGSTHEPLASMACONCENTRATIONTIMECURVESWEREPLOTTEDANDTHEPHARMACOKINETICPARAMETERSWEREESTIMATEDANDANALYZEDRESULTSDIFFERENCESINPEAKTIMETMAX,ELIMINATIONHALFTIMET1/2BETWEENBULKDRUGANDSUSTAINEDRELEASEPELLETSHAVEBEENFOUNDSTATISTICALLYSIGNIFICANTP005,ANDTHEPEAKCONCENTRATIONCMAXOFBULKDRUGWASHIGHERTHANSUSTAINEDRELEASEPELLETSAFTERADOSECONVERSIONP005CONCLUSIONSCOMPAREDWITHTHEBULKDRUGOFTAMSULOSINHYDROCHLORIDE,THERATESOFABSORPTIONANDELIMINATIONWERELONGER,ANDTHEPLASMAPEAKCONCENTRATIONWASLOWERINSUSTAINEDRELEASEPELLETS,WHICHIMPROVEDASUSTAINEDRELEASEPERFORMANCEOFTHISPREPARATIONKEYWORDSTAMSULOSINHYDROCHLORIDESUSTAINEDRELEASEPELLETSLC/MS/MSPHARMACOKINETICS2介绍盐酸坦洛新，化学名为R5222乙氧苯基乙醚基胺基2甲基乙基2甲氧基苯磺酰胺盐酸盐，属肾上腺素1G2475体G1134G34231A的G19471断剂，对G4627G17959、G14164G14021G20060G18108及G2081G2027腺存在的1G2475体具有G20652G17885G6333性G19471断G1328用，G1186G13792降低G5191G9381G13920G5364G2159，降低G991G4627G17347G19471G2159，G1032G5214用G1122G8847G11115G2081G2027腺G3698G10995G6164G14280的异G5132G6502G4627G11163G10378，G3926G4627G20069、G3824G4627G3698G3822、G6502G4627G3268G19602G125731。G11013G1122坦洛新在体内消除G17817速，G10628有上G5078G2842G2709均为缓释制剂，G3926盐酸坦洛新缓释G14026G3230、缓释G10267G12573。G7424G16809G20576G11458的在G1122G1209盐酸坦洛新原料药为对G10043，G1209BEAGLE犬为G2475G16809动物，评价盐酸坦洛新缓释微丸在体内的缓释特征。SOH3CH2NOOHNOCH3OH3C图1坦洛新化学G13479G7512G5347FIG1CHEMICALSTRUCTUREOFTAMSULOSIN1材料与仪器11药G2709与G16809剂盐酸坦洛新原料药G6221G250739060901，G13443度992，盐酸坦洛新缓释微丸G6221G250708042491，G8611188MGG2559盐酸坦洛新02MG，均G11013G8755G14499G5670G10802G2319药G13941G1233有G19492G1856G2508G6564G1391。内G7643物SHR117616G6221G250720070920，G13443度953，G11013G7424G1856G2508化学合G6116组合G6116。HPLC甲G18267G17153G14270THERMOFISHERG1856G2508，G8712为G4636G14263G8675G14988G20323G8712，其G1325G16809剂为分析G13443，G17153G14270G3281药G19610G3254化学G16809剂有G19492G1856G2508。12G16809G20576动物G1593G5259G6116G5192BEAGLE犬8G2494，G19616G19608各半，体G1833768KG，G11013G14499G5042中G12197G4466G20576动物有G19492G1856G2508G6564G1391，动物G10995G1147G16780G2499G16789G2507G726SYXKG1449920020032。13G1214G3132与G16786G38033TSQQUANTUMULTRAAMG989G18337G3247G7509G7450G1030G13864G17148G16901G1214，G13666G3281THERMOFINNIGANG1856G2508；AGILENT1200G20652G6940G9094相G14406G16901G13007统，德G3281AGILENTG1856G2508。XCALIBAR50数据处理软件及DAS20药代动G2159学软件。2试验部分21G16809G20576方案G1593G5259G6116G5192BEAGLE犬8G2494，G5191均分G61162组，禁食过G3824后分别口服给予盐酸坦洛新原料药及其缓释微丸灌制的G14026G3230，给药剂量分别为1MGKG1和2MGKG1。原料药组G1122给药G2081及给药后1H,2H,3H,4H,5H,6H,8H,10H,12H,24H,36HG11013G2081肢静脉采血15ML，置G1122肝素化G16809管中，3500RPM离心10MIN分离血浆，G112220C保存。缓释微丸组G1122给药G2081及给药后15MIN,30MIN,45MIN,1H,2H,3H,4H,6H,8H,12H,24HG11013G2081肢静脉采血15ML，处理方法同原料药组。22LC/MS/MS法测定犬血浆中的坦洛新参G10043有关文献24，建立了快速、灵敏的LC/MS/MS法用G1122测定犬血浆中的坦洛新浓度，样G2709经碱化、G9094G9094G6564取后进行分析。221分析条件G14406G16901条件G726G14406G16901柱为HYPERSILC18柱150MM46MM,5M，保护柱为EASYGUARDC18柱10MM46MM,5M，流动相为甲G182672甲酸G8712溶G90947030,V/V，流速06MLMIN1，柱温25C。G17148G16901条件G726ESI源，正离子扫描，检测方G5347为G17885G6333反应监测SRM，鞘气压G215960PSI，辅助气15PSI，毛细管温度300C。监测反应为坦洛新M/Z40922280CE26EV，内G7643SHR117616M/Z54023670CE25EV。222血浆样G2709预处理取犬血浆200L，混合后加入内G7643溶G909450L1000NGML1SHR117616甲G18267溶G9094，加入甲G18267G726G87128020,V/V50L，饱和NA2CO3溶G9094100L；混匀后加入无G8712乙醚4ML，涡流混合2MIN，振荡10MIN，离心10MIN4000RPM；取上清G9094G112240C空气流G991吹干，残留物溶G1122100L流动相，取10L进行LCMSMS分析。223方法学确G16789按G10043G10995物样G2709分析方法确G16789指导原则的有关要求5，对建立的分析方法进行4了方法学确G16789。方法的G17885G6333性G13479果见图2。G13479果表明，血浆中的内源性物G17148不干扰待测物坦洛新及内G7643物SHR117616的测定。图2LC/MS/MS法测定犬血浆中坦洛新的典G3423G14406G16901图A空白血浆样G2709；B空白血浆加入02NGML1坦洛新及250NGML1内G7643；C一G2494犬口服给予1MGKG1原料药后1H的血浆样G2709峰I坦洛新；峰II内G7643SHR117616FIG2REPRESENTATIVELC/MS/MSCHROMATOGRAMSOFTAMSULOSININPLASMAOFABCIII001020304050TIMEMIN020406080100020406080100RELATIVEABUNDANCENL750E1SRMMS24092026002279522805NL232E2SRMMS25402025003669536705RELATIVEABUNDANCE001020304050TIMEMIN020406080100020406080100396446NL453E3SRMMS24092026002279522805NL238E5SRMMS25402025003669536705RELATIVEABUNDANCERELATIVEABUNDANCE001020304050TIMEMIN020406080100020406080100RELATIVEABUNDANCE396445NL122E4SRMMS24092026002279522805NL246E5SRMMS25402025003669536705RELATIVEABUNDANCEIII5BEAGLEDOGSABLANKPLASMABBLANKPLASMASPIKEDWITH02NGML1TAMSULOSINAND250NGML1INTERNALSTANDARDCPLASMASAMPLEOFABEAGLEDOG1HAFTERANORALADMINISTRATIONOF1MGKG1TAMSULOSINPEAKITAMSULOSIN；PEAKIIINTERNALSTANDARDSHR117616取犬空白血浆200L，加入坦洛新G7643准G13007G2027溶G909450L，使血药浓度分别为0200,0500,100,500,100,250,500NGML1，加入内G7643溶G909450L，G14270加入饱和NA2CO3溶G9094100L起，按“血浆样品预处理”项G991进行操G1328。G1209血药浓度为横坐G7643，样G2709与内G7643G14406G16901峰面积比为纵坐G7643，G1209加权最小二乘法W1/X2进行线性回归，获得典G3423G7643准曲线。G13479果表明，该方法在0200500NGML1浓度范围内线性良好，最低定量G194920200NGML1。典G3423的G7643准曲线方程为Y006635X000276，R209948。G12090500,500,450NGML1为G17148控样G2709，对方法的精密度与准确度、G6564取回收率、稳定性G12573分别进行了考察，G13479果均符合相关规范的要求5，且坦洛新G7643准溶G9094、血浆样G2709G1122室温G991放置12H、4CG991放置7G3837均保G6357稳定。3试验结果31血浆药物浓度测定采用建立的LC/MS/MS方法对BEAGLE犬口服给予盐酸坦洛新及其缓释微丸后各时刻的血浆药物浓度进行测定，绘制的血浆药物浓度时间曲线见图3。0501001502002500060120180240300360TIMEHCONCENTRATION/NGML1图3BEAGLE犬分别口服给予盐酸坦洛新及其缓释微丸后的G5191均血药浓度时间曲线N46G441原料药；G442缓释微丸FIG3MEANPLASMACONCENTRATIONTIMECURVESOFTAMSULOSININBEAGLEDOGSAFTERANORALADMINISTRATIONOFTAMSULOSINHYDROCHLORIDEANDITSSUSTAINEDRELEASEPELLETSN4G441BULKDRUG；G442SUSTAINEDRELEASEPELLETS32药动学参数对血药浓度时间数据采用DAS20药动学软件进行G6163室G8181G3423G6323合并计算主要药动学参数，其中TMAX、CMAX采用G4466测G1552。G13479果表明，盐酸坦洛新及其缓释微丸在BEAGLE犬体内的药动学行为符合二室G5332放G8181G3423权G18337G13007数W1，主要药动学参数见表1。表1BEAGLE犬口服给予盐酸坦洛新及其缓释微丸后的主要药动学参数TAB1MAINPHARMACOKINETICPARAMETERSOFTAMSULOSININBEAGLEDOGSAFTERANORALADMINISTRATIONOFTAMSULOSINHYDROCHLORIDEANDITSSUSTAINEDRELEASEPELLETSPARAMETERSBULKDRUGSUSTAINEDRELEASEPELLETSTMAX/H144105450129CMAX/NGML1267133949524T1/2/H113039256050T1/2/H032019148042T1/2/H108054307235AUC0T/NGHML1417174511296AUC0A1/NGHML1419173511296MRT0A1/H236125622100VZ/LKG1459268101674CLZ/LH1KG127612225814933统计学分析对原料药及缓释微丸给药组主要药动学参数进行统计学分析，G13479果表明G726BEAGLE犬口服给予盐酸坦洛新原料及缓释微丸后，血浆中药物达峰时间TMAX、消除半衰期T1/2均存在显著性差异P005；两组峰浓度CMAX经剂量折算后存在显著性差异P005。4讨论7BEAGLE犬口服盐酸坦洛新原料后，吸收G17817速，消除半衰期G11713；与盐酸坦洛新原料药相比，BEAGLE犬口服缓释微丸后，吸收及消除速度减慢、血药峰浓度降低，符合缓释制剂的药动学行为，判断该制剂具有一定程度的缓释特征。盐酸坦洛新G3252体内消除速度过快，G1032G5214均采用缓释制剂G1209G5322缓其消除，G5322G19