This study aimed to characterize and predict the pharmacokinetic (PK) properties of CU05-1189, an established PDK1 PH domain inhibitor with anti-angiogenic effects demonstrated by suppression of VEGF-induced angiogenesis in xenograft models. A validated bioanalytical method (0.01–10 μg/mL, precision/accuracy within ±15%) enabled CU05-1189 plasma quantification. A one-compartment PK model was established using NONMEM software to characterize and predict plasma concentration-time profiles. The established PK model effectively captured the observed plasma concentration data, with most values within the 90% confidence interval. The predicted clearance values from multiple-dose administration fell between those observed for single-dose and repeated-dose studies. Notable PK parameters at the 100 mg/kg dose included a maximum concentration(Cmax) of 85.676 ± 24.484 μg/mL, area under the curve(AUClast) of 270.964 ± 119.955 μg·h/mL, and an elimination half-life of approximately 8 hours. Furthermore, dose-proportional PK was confirmed across the tested dose range. These findings provide insights into CU05-1189 pharmacokinetics, supporting human plasma concentration predictions. The developed PK model will be instrumental in extrapolating human PK profiles and optimizing dose selection for preclinical and clinical development.