The Non-clinical Pharmacokinetic Study Of HZZ112 In Rodents

HZZ112 is a developed novel anti-cancer drug candidate. According to the "Guideline for the non-clinical pharmacokinetic study of chemical drugs" requirement, this paper focuses on the pharmacokinetics in rat plasma, tissue distribution and excretion of HZZ112 and its active metabolites; also the differences in the pharmacokinetics of HZZ112 in normal nude mice and nude mice bearing tumor. Through the acquisition of scientific data and comprehensive analysis and evaluation, we got the pharmacokinetic characteristics of HZZ112 and its metabolites, including absorption; the urine and fecal excretion; the extent of drug accumulation in the body and major organ. These results provided valuable information for the full evaluation of HZZ112.1. Pharmacokinetic studies on HZZ112 and its metabolites in rat plasmaObjective To study the pharmacokinetic characteristics of HZZ112 and its two major active metabolites following i.v. injection in three different doses.Methods A simple and sensitive method for the determination of HZZ112 and its metabolites in rat plasma was developed using high performance liquid chromatography (HPLC). The pharmacokinetic parameters of HZZ112 and its metabolites in rat were fitted using DAS 2.0 software.Results The calibration curves showed good linear for HZZ112 and its metabolites. For HZZ112, assay recoveries ranged from 90.5% to 114.5%, both the intra-and inter-day precisions were less than 10.7%. For HZZ1005, assay recoveries ranged from 87.3% to 106.9%, both the intra- and inter-day precisions were less than 8.5%. For HZZ1018, assay recoveries ranged from 89.2% to 107.9%, both the intra-and inter-day precisions were less than 6.6%. The stability at different conditions met the requirements for biopharmaceutical analysis. After i.v. injection of HZZ112 in doses of 7,28, and 56 mg/kg body weight in rat, the AUC(o-t) of HZZ112 were 58.9±23.0,224.7±117.0,470.2±316.7 mg/L*h, respectively; the t1/2 were 0.45 h, 3.1,1.1 h, respectively; the Cmax were 111.9±14.0,116.4±69.2,259.3±130.3μg/mL, respectively. The AUC(0-t)of HZZ1005 were 0.736±0.44,1.02±0.16,2.13±.13 mg/L*h, respectively; the t1/2 was 2.0 h; the Cmax were 0.32±0.09,0.48±0.08,1.02± 0.35μg/mL, respectively.The AUC(0-t) of HZZ1018 were 0.109±.027,0.356±0.14, 0.66±0.19 mg/L*h, respectively; the t1/2 were 1.0,0.74,1.4 h; the Cmax were 0.18± 0.03,0.55±0.19,0.61±0.26 μg/mL, respectively.Conclusion HZZ112 and its metabolites have good linear relationships between their Cmax and AUC(0-t) and their administrated doses. The AU(0-t) of metabolites are less than 10% of that of the prototype.2 Studies on the distribution of HZZ112 and its metabolites in ratsObjective To investigate the distribution of HZZ112 and its metabolites in rat heart, liver, spleen, lung, kidney, brain, muscle, small intestine, stomach, pancreas, thymus, fat, testis or uterus, ovary tissues after i.v. injected HZZ112 at a dose of 28 mg/kg body weight in rat.Methods A simple and sensitive method for the determination of HZZ112 and its metabolites in rat tissue was developed using HPLC. The tissues were taken out at 0.167,2, and 5 h after administrated HZZ112. Then the concentration of HZZ112 and its metabolites in tissues were determined after the samples processed.Results After i.v. administrated, HZZ112 mainly existed in the heart, liver, spleen, lung and fat tissue at 0.167 and 2.0 h, and mainly existed in the spleen and fat at 5 h; HZZ1018 mainly existed in the liver, spleen, kidney, lung and pancreas at 0.167 and 2.0 h; HZZ1005 mainly existed in the liver, spleen and pancreas at 0.167 and 2.0 h.Conclusion HZZ112 and its metabolites were widely distributed in rat tissues, and no significant drug accumulation phenomenon was found.3 Urinary and fecal excretion study of HZZ112 and its metabolitesObjective To study urinary and fecal excretion in rat after i.v. injection of HZZ112 at a dose of 28 mg/kg body weight.Methods A rapid and sensitive method for the determination of metabolites in rat urine and feces was developed using HPLC. The urine after i.v. injection of HZZ112 were compared with blank urine and hydrolyzed to observe HZZ112 and its potential metabolites in urine. Urine and feces were collected after i.v. injection of HZZ112 at a dose of 28 mg/kg within 60 h and were quantitatively determination by HPLC. The total excretion rate was calculated.Results HZZ112 was mainly excreted as its active metabolites in rat urine and feces. The calibration curves both showed good linears for HZZ1005 and HZZ1018 in rat urine and feces. HZZ1005 extraction recoveries ranged from 90.4% to 100.0%; assay recoveries ranged from 86.5% to 98.1%; both the intra- and inter-day precisions were less than 9.5% in urine. HZZ1018 extraction recoveries ranged from 89.9% to 111.0%;, assay recoveries ranged from 85.6% to 107.4%; both the intra- and inter-day precisions were less than 7.5% in urine. HZZ1005 extraction recoveries ranged from 57.8% to 77.6%; assay recoveries ranged from 83.9% to 101.7%; the intra-day precision was less than 7.1% in feces. HZZ1005 extraction recoveries ranged from 80.6% to 97.0%; assay recoveries ranged from 86.4% to 106.5%; the intra-precision was less than 8.4%. Conclusion HZZ112 was excreted mainly in the form of metabolites in rat urine and feces. The total excretion rate of HZZ112 within 60 h after i.v. injection was 24.7± 5.7%(n=6).4 The pharmacokinetic study of HZZ112 and its metabolites in nude miceObjective To compare the pharmacokinetic differences of HZZ112 in normal nude mice and nude mice bearing tumor after i.v. injection of HZZ112 at a dose of 40 mg/kg body weight.Methods A simple and sensitive method for the determination of HZZ112 and its metabolites in mice plasma and tumor tissue was developed using HPLC. The concentrations of plasma and tissue samples were determined. Then the concentration-time curve and pharmacokinetic parameters were obtained.Results After i.v. injection of HZZ112, the AUC(0-t) of HZZ112 were 555.3 and 376.1 mg/L*h in normal nude mice and mice bearing tumor, respectively; the t1/2 were 0.865 and 0.841 h, respectively; the Cmax were 580.8 and 482.6 μg/mL, respectively. The AUC (0-t) of HZZ1005 and HZZ1018 were both less than 10% of HZZ112. Tumor tissues contained only trace amounts of HZZ112, and its metabolites were not detected.Conclusion There were no significant pharmacokinetic differences between normal and tumor-bearing nude mice after i.v. administration of HZZ112.