Quantitative Analysis of Daporinad (FK866) and Its In Vitro and In Vivo Metabolite Identification Using Liquid Chromatography-Quadrupole-Time-of-Flight Mass Spectrometry

Abstract

Daporinad (FK866) is one of the highly specific inhibitors of nicotinamide phosphoribosyl transferase (NAMPT) and known to have its unique mechanism of action that induces the tumor cell apoptosis. In this study, a simple and sensitive liquid chromatography–quadrupole-time-of-flight–mass spectrometric (LC-qTOF-MS) assay has been developed for the evaluation of drug metabolism and pharmacokinetics (DMPK) properties of Daporinad in mice. A simple protein precipitation method using acetonitrile (ACN) was used for the sample preparation and the pre-treated samples were separated by a C18 column. The calibration curve was evaluated in the range of 1.02~2220 ng/mL and the quadratic regression (weighted 1/concentration2) was used for the best fit of the curve with a correlation coefficient ≥ 0.99. The qualification run met the acceptance criteria of ±25% accuracy and precision values for QC samples. The dilution integrity was verified for 5, 10 and 30-fold dilution and the accuracy and precision of the dilution QC samples were also satisfactory within ±25% of the nominal values. The stability results indicated that Daporinad was stable for the following conditions: short-term (4 h), long-term (2 weeks), freeze/thaw (three cycles). This qualified method was successfully applied to intravenous (IV) pharmacokinetic (PK) studies of Daporinad in mice at doses of 5, 10 and 30 mg/kg. As a result, it showed a linear PK tendency in the dose range from 5 to 10 mg/kg, but a non-linear PK tendency in the dose of 30 mg/kg. In addition, in vitro and in vivo metabolite identification (Met ID) studies were conducted to understand the PK properties of Daporinad and the results showed that a total of 25 metabolites were identified as ten different types of metabolism in our experimental conditions. In conclusion, the LC-qTOF-MS assay was successfully developed for the quantification of Daporinad in mouse plasma as well as for its in vitro and in vivo metabolite identification.

Interpretable Drug Response Prediction through Molecule Structure-aware and Knowledge-Guided Visible Neural Network

“…Daporinad, Docetaxel and Bortezomib are the top-3 sensitive drugs, which is consistent with the prediction of previous DRP methods [23,24,35]. The most sensitive drug, Daporinad, is one of the highly specific inhibitors of nicotinamide phosphoribosyl transferase and known to have its unique mechanism of action that induces the tumor cell apoptosis [36]. Among the prediction results of Daporinad, (Daporinad, MOLM-13) has the lowest value of -8.82.…”

Channeling Nicotinamide Phosphoribosyltransferase (NAMPT) to Address Life and Death

“…After the termination of infusion, the drug rapidly clears . Delivered iv to mice, T 1/2 ≈ 50 min, C max = 14 μM (at 10 mg/kg) and 25 metabolites were identified including the N -oxide that was also measured in human trials . Although the activity of the N -oxide metabolite has not been reported, rapid clearance likely translates to little or no contribution to the in vivo activity of FK866.…”

“… 38 Delivered iv to mice, T 1/2 ≈ 50 min, C max = 14 μM (at 10 mg/kg) and 25 metabolites were identified including the N -oxide that was also measured in human trials. 39 Although the activity of the N -oxide metabolite has not been reported, rapid clearance likely translates to little or no contribution to the in vivo activity of FK866. In toxicodynamic studies in rats, FK866 (ip up to 60 mg/kg bd) in plasma reached C max = 6.7 μM.…”