Studies On Drug-Drug Interactions Of Xiaoaiping Injection

Xiaoaiping injection, a standard Marsdenia tenacissima (MTE), is made from the extracts of the stem of Marsdenia tenacissima (Roxb.) Wight et Am.(Asclepiadaceae), known as "Tong-guan-teng" in Chinese folk medicine. Traditionally, it has the efficacy of detoxify, Huatanruanjianpian, etc., mainly used in clinical main treatment for lung cancer, liver cancer, esophageal cancer, stomach cancer and other malignancies with good clinical efficacy, the addition xiaoaiping injection combined with radiotherapy and chemotherapy for advanced nasopharyngeal cancer, ovarian cancer, lymphoma of the oldest age.Xiaoaiping injection has been widely used in clinical practice, and there are many pharmacological effects and clinical applications investigations reported about xiaoaiping injection, however, its pharmacokinetic characteristics of the study is relatively scarce. Therefore, understanding the xiaoaiping injection in clinical practice, often in combination with other drugs, may cause adverse reactions. This paper attempts to be determined by the content of the xiaoaiping injection, based on the main ingredient in the impact and interaction of cytochrome P450and injection pharmacokinetic study, xiaoaiping injection in clinical medication safety reference.Chapter1Simultaneous Determination of Seven Major compound in Xiaoaiping Injection by HPLCObjective:To develop an HPLC method for the simultaneous determination of seven major compound in Xiaoaiping injection. Methods:The analytical column was Kromasil100-5C18(4.6x250mm,5μm). The mobile phase was acetonitrile-0.2%phosphoric acid, gradient elution. The flow was0.8mL·min-1. The detection wavelength was300nm. The column temperature was30℃. Results:The linear ranges of seven compounds were as fllows:0.1-3.2mg-L-1(r=0.9993) for neochlorogenic acid,0.025～0.8mg·L-1(r=0.9999) for Protocatechuic aldehyde,0.1～3.2mg·L-1(r=0.9998) for chlorogenic acid,0.075～2.4mg·L-1(r=0.9999) for4-dicaffeoylquinic acid,0.0375～1.2mg·L-1(r=0.9999) for vanillic acid,0.02～0.64 mg·L-1(r=0.9999) for caffeic acid,0.01-0.32mg·L-1(r=0.9999)for4-coumararate.The average recoveries were99.6%(RSD0.24%),100.0%(RSD0.15%),98.7%(RSD1.00%),99.1%(RSD1.36%),96.4%(RSD1.37%),98.3%(RSD1.80%),97.3%(RSD1.64%) respectively. Conclusion:This method is simple, accurate and available for the quality control of Xiao aiping Injection.Chapter2Simultaneous Determination of Two Activitive compound in Xiaoaiping Injection by LC-MSObjective:A rapid and specific high performance liquid chromatography-mass spectrometric method was developed for determination of17β-tenacigenin B and tenacigenoside A in Xiaoaiping injection. Methods:The analytical column was Agilent Zorbax SB-C18(3.0×100mm,3.5μm) and the security guard column was Agilent Zorbax SB-C18(2.1×12.5mm,5μm). The mobile phase was methanol-water(90:10) at a flow rate of0.25mL·min-1. Methyltestosterone as internal standard, electrospray ionization model (ESI), and SIM model were used for quantification. Results:The linear ranges of two compounds were as fllows:0.625-20mg·L-1(r=0.9983) for17β-tenacigenin B,0.375～12mg·L-1(r=0.9994) for tenacigenoside A. The average recoveries were100.8%(RSD3.95%),101.6%(RSD2.71%), respectively. Conclusion:This established method was highly sensitive, fast, accurate and available for the quality control of Xiaoaiping Injection.Chapter3Validation of an LC-MS method for the simultaneous quantification of two bioactive constituents in rat plasma after administration of Xiaoaiping InjectionAim of the study:A LC-MS method was developed and validated for the simultaneous quantification of two bioactive constituents in rat plasma after administration of Xiaoaiping Injection. Methods:Chromatographic separation was Agilent Zorbax SB-C18(3.0×100mm,3.5μm) and the security guard column was Agilent Zorbax SB-C18(2.1×12.5mm,5μm). The mobile phase was methanol-water(90:10) at a flow rate of0.25mL·min-1. Eight rats were administered with Xiaoaiping Injection, and LC-MS was utilized to simultaneously quantitate2bioactive compounds in rat plasma, and DAS1.0Software was used to fit plasma concentration-time curve and calculate their corresponding principal pharmacokinetic parameters. Results:The calibration curves of17β-tenacigenin B and tenacigenoside A revealed good linear regression (r≥0.996) within test ranges. This method provided excellent sensitivity with good precision with RSDs of intra-and inter-day variation less than5.7%and5.3%, respectively. Conclusion:The validated method was then applied for pharmacokinetic study of the2constituents in rat plasma after administration of Xiaoaiping Injection.Chapter4Effects of the Xiaoaiping injection on Liver S9in MiceObjective:To investigate the effect of Xiaoaiping injection on activities of total CYP450. Methods:20mice were divided into four groups. One is control group (CG), the second is positive control group and the third and forth groups are administed Xiaoaiping injection groups. The mice of CG were given saline only and the mice of positive control group were given Phenobarbital sodium injection (PBN) at80mg·kg-1by peritoneal injection (i.p.), once daily for ten days. The remaining two groups were i.p. with6mL·kg-1(normal-dosage group, NG) and12mL·kg-1(high-dosage group, HG) Xiaoaiping injection, respectively, once daily for seven days. Ultraviolet-visible spectrometry (UV) has been applied to determine contents of hepatic S9protein at660nm, CYP450at450nm. The statistical differences of liver coefficient, hepatic S9protein content, CYP450content between dosing group mice (normal-dose group and higher-dose group)and control group mice (physical saline group and phenobarbital group) were evaluated by One-way Analysis of Variance (One-way ANOVA). Results:Two doses of Xiaoaiping injection had no significant impact on mice liver coefficient and protein content (P>0.05),but high-dosage group and normal-dose group also might increase CYP450content. Conclusion: Normal-dose group and high doses of Xiaoaiping injection could also induce activity CYP450.Chapter5Investigation of Xiaoaiping Injection’s Impacts on the P450Activities Using a Cocktail MethodObjective:To develop a liquid chromatography-tandem mass spectrometry method for simultaneous determination of six major cytochrome P450probe durgs, theophylline (CYP1A2), tolbutamide (CYP2C9), omeprazole (CYP2C19), dextromethorphan (CYP2D6), chlorzoxazone (CYP2E1) and midazolam (CYP3A4), and to evaluate the effects of Xiaoaiping injection on P450activities by cocktail probe drugs in rats. Method:8rats underwent2-cycle pharmacokinetic experiments before and after being treated with clinically-dose Xiaoaiping injection for10days, in which the rats were concomitantly administered tolbutamide (20mg·kg-1), chlorzoxazone (40mg·kg-1), theophylline (25mg·kg-1), midazolam (20mg·kg-1), omeprazole (40mg·kg-1), and dextromethorphan (30mg·kg-1) by gastrogavage, followed by blood-withdrawing from orbital bleeding at different intervals with24hours. LC-MS/MS was utilized to simultaneously quantitate6probe compouds in rat plasma, and DAS1.0Software was used to fit flasma concentration-time curve and calculate their corresponding principal pharmacokinetic parameters, among which the statistical differences were evaluated by Paried t-test. Results:The2-cycle pharmacokinetic parameters of theophylline for clinically-dose rats exhibited insignificant differences (P>0.05) meanwhile, after being treated with clinically-dose Xiaoaiping injection, but AUCo-24h of tolbutamide after treatment was lower than before treatment (P<0.05), and AUCo-24h of omeprazole, dextromethorphan, chlorzoxazone, midazolam after treatment was larger than before treatment (P<0.05). Conclusion:In the10-day administration period, clinically-dose Xiaoaiping injection could not insignificantly change CYP1A2activity, but could inhibit activities of CYP3A4, CYP2C19, CYP2D6, CYP2E1and induce CYP2C9.Chapter6Effects of Xiaoaiping injection on OndansetronHydrochloride pharmacokinetics in ratsAim of the study:This study was aimed investigating the effect of Xiaoaiping injection on pharmacokinetics of ondansetron in rats. Methods:Eight rats were administered with Xiaoaiping injection (6mL·kg-1) once daily for10consecutive days. All the rats were administered orally with the ondansetron(4mg·kg-1) before the first time and after the last time given Xiaoaiping injection. HPLC was utilized to determine the concentration of ondansetron in rat plasma and to calculate the corresponding pharmacokinetic parameters. The statistical differences of the two cycles were evaluated by paired-samples t-test. Results:In the rats treated with Xiaoaiping injection and ondansetron, the t1/2of ondansetron increased obviously (p<0.05) compared with the ondansetron alone, while maximum plasma concentrations (Cmax), area under the plasma concentration-time curve (AUC(0-t)) decreased significantly (p<0.05). There was no significant difference between other parameters. Conclusion:Xiaoaiping injection can reduce the absorption of ondansetron and accelerate the metabolism of ondansetron.