| Making use of probe method to measure the activity of CYP3A4 in this study,and research 9 kinds of anti-tumor traditional Chinese medicine injection,which include cinobufacini?kangai?lentinan?xiaoaiping?kanglaite?aidi?elemene?shenfu and brucea jananica oil injection.According to the regulation of the CYP450 enzyme,which can be predicted by the interaction of CYP3A4 metabolism of anti-tumor drugs with the 9 kinds of traditional Chinese medicine injection for adjusting the clinical medication plan,improving treatment effect,reducing adverse reaction,and realizing individualized medication are of significance.Therefore,the main work of this study includes four aspects of the following two chapters:Chapter 1 Influence of 9 anti-tumor Chinese medicine injections on the activitise of cytochrome P450 isoforms CYP3A4 in rat liver microsomes incubation in vitroObjective: To invesigate the influence of cinobufacini?kangai?lentinan?xiaoaiping?kanglaite?aidi?elemene?shenfu and brucea jananica oil injection on CYP3A4 of liver microsomes in vitro as probe.Methods: Chromatographic conditions: column of chromatography: German MN Nucleodur C18 column(5 ?m,4.6 mm x250 mm);mobile phase: acetonitrile water(v:v=45:55);detection wavelength: 245 nm;flow rate: 1 ml/min;column temperature: 30?.Testosterone was used as a probe of CYP3A4 in rat liver microsomes in vitro incubation,HPLC method was used to detect the production of 6?-Hydroxytestosterone(6?-OHTS)to study the effects of 9 anti-tumor traditional chinese medicine Injections on the activities of CYP3A4 enzymes in the microsomal incubation system.Results: The concentration of rat liver microsomes is 0.4 mg/m L,testosterone solution is 50 mol/L,and the optimal incubation time is 30 min.Pharmacokinetic study: in the liver microsome incubation system,the linear equation of6-OHTS is Y=0.0608X+0.0102(r=0.9987),the linear range is 0.075~15?mol/L,the linear relationship is good,LOD is 0.002 ?mol/L.The intra-day and inter-day precision and of RSD are less than 7%,RME are less than 2%,the recovery rate of RSD is less than 6%,and the stability of RSD is less than 7%.The influence of CYP3A4 on the activities of enzyme are compared with the blank control group,low dose group:inhibiting the activities of CYP3A4 enzymes' drugs are kangai ?xiaoaiping and lentinan,induction of drugs include cinobufacini ?shengfu?kanglaite and brucea jananica oil,elemene and aidi have no obvious induction or inhibition;middle dose group: drug induced influence on CYP3A4 enzyme activities of cinobufacini ? aidi and kanglaite,inhibition of drugs include kangai?lentinan?xiaoaiping and brucea jananica oil,shenfu and elemene have no obvious induction or inhibition;high dose group: inhibiting the activities of CYP3A4 enzymes' drugs are kangai?lentinan?xiaoaiping?aidi?kanglaite and brucea jananica oil,cinobufacini has induction,shenfu and elemene have no obvious induction or inhibition.Brucea javanica oil on CYP3A4 enzyme's half inhibitory concentration IC50 is 0.78%,the xiaoaiping half inhibitory concentration IC50 is 0.69%.Conclusions: In clinical,when the anti-tumor traditional Chinese medicine injections and the use of CYP3A4 drugs combined with radiotherapy or chemotherapy,need to be wary of the anti-tumor traditional Chinese medicine injection(kanglaite?aidi?brucea javanica oil?cinobufotalin?lentinan?kangai and xiaoaiping)and between the drugs for potential drug interactions.Chapter 2 pharmacokinetic study of xiaoaiping and docetaxel in vivo of ratsObjective: To establish a HPLC method for determination of docetaxel in rats,the influence of docetaxel pharmacokinetic of xiaoaiping in vivo of rats,and to provide experimental basis for clinical rational drug use.Methods: Chromatographic conditions: column of chromatography: German MN Nucleodur C18 column(5 ?m,4.6 mm x250 mm);mobile phase: acetonitrile water(v:v=50:50);detection wavelength: 233 nm;flow rate: 1 ml/min;column temperature: 25?.SD rats are randomly divided into 4 groups,the rats are injected the different drugs which according to the group and weight of rat,except for the A group only injected docetaxel and a small amount of saline,B?C?D groups of rats are injected in docetaxel,a small amount of saline,and the low?middle and high doses of xiaoaiping.Basis for the set time points to get bloods,blood concentration of docetaxel is determined by HPLC method.The pharmacokinetic parameters are fitted by DAS2.1.1 software,and the parameters were analyzed by SPSS 17 software.Results:Pharmacokinetic study: in the liver microsome incubation system,the linear equation of 6-OHTS is Y=8.1013X-3.2652(r>0.999),the linear range is 0.7813~75 ?g/ml,the linear relationship is good,LOD is 0.5208?g/L.The intra-day and inter-day precision RSD of docetaxel is less than8%,RME is less than 4%;the recovery rate of docetaxel RSD is less than7%;the stability under the different conditions of the RSD are less than8%.In terms of pharmacokinetics,4 groups of Plasma concentration are fitted by DAS2.1.1 software,and the main pharmacokinetic parameters included:A group: AUC0?t(60.22±3.65?g/ml·h)? AUC0??(69.93±5.39?g/ml·h)?Vd(0.18±0.04L/kg)?CL(0.19±0.01ml/kg·h-1)?t1/2(?)(3.08±1.21h);B group: AUC0?t(62.29±3.65?g/ml·h)?AUC0??(72.44±7.23?g/ml·h)?Vd(0.18±0.03L/kg)?CL(0.22±0.02ml/kg·h-1)?t1/2(?)(3.95±1.39h);C group: AUC0?t(70.58±11.62?g/ml·h)?AUC0??(89.45±19.43?g/ml·h)?Vd(0.16±0.05L/kg)?CL(0.16±0.04ml/kg·h-1)?t1/2(?)(4.53±1.77h);D group: AUC0?t(97.67±20.40?g/ml·h)?AUC0??(109.32±41.46?g/ml·h)? Vd(0.14±0.04L/kg)? CL (0.15±0.07ml/kg·h-1)?t1/2(?)(5.72±2.26h).Compared with group A,AUC of the other three groups are significantly prolonged,increased bioavailability of docetaxel,C and D group is statistically significant;the t1/2(?)of the other three groups are significantly prolonged,docetaxel in rats in vivo elimination slowed,but only D group was statistically significant.Except for B group,the remaining of two groups' CL are decreased,reducing the elimination of docetaxel in rats,but only D is statistically significant,no significant remaining all parameters.Conclusions: The HPLC method is quick and simple,flexible,and can be used for the pharmacokinetic study of docetaxel;after combined use of docetaxel and xaoaiping,multiple pharmacokinetic parameters of docetaxel have obviously changed,there is an interaction between this two drugs,xiaoaiping has obvious inhibitory effects on docetaxel.The results of chapter 2 not only validate the conclusion of the chapter 1 in this thesis,but also be aware of the potential drug interactions between the two types of drugs,especially for radiotherapy or chemotherapy. |
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