Studies on the metabolism of 4-methyl-piperazine-1-carbodithioc acid 3-cyano-3,3-diphenylpropyl ester hydrochloride in rats by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry.

Department of Pharmaceutical Analysis, School of Pharmaceutical Sciences, Peking University, Xueyuan Road 38, Beijing 100083, China.
Journal of Pharmaceutical and Biomedical Analysis (Impact Factor: 2.83). 10/2007; 44(5):1127-32. DOI: 10.1016/j.jpba.2007.05.026
Source: PubMed

ABSTRACT 4-Methyl-piperazine-1-carbodithioc acid 3-cyano-3,3-diphenylpropyl ester hydrochloride(TM208) is a newly synthesized compound, which has shown excellent in vivo and in vitro anticancer activity and low toxicity. In this study, the metabolism of TM208 in rats was studied for the first time by high-performance liquid chromatography coupled with tandem mass spectrometry. Following a single oral administration to rats, TM208 was metabolized to eight metabolites (M1-M8). M1 is the desmethyl metabolite and the acylation of M1 with N-acetyl transferase results in M6 (N-acetyl metabolite), M5 is N-formyl metabolite; M4 is phenyl monohydroxylation metabolite, M2 is the sulfine metabolite of TM208, and M3 is also an odd-oxygen added products which the possible oxidation site has described in this paper; M8 is the metabolite resulting from the replacement of '-C=S' with '-C=O', M7 is a ring-opened piperazine oxidation products to a kind of acid.

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    ABSTRACT: A new, specific and sensitive high performance liquid chromatography analytical procedure was developed and validated for the determination of TM208 in rat primary solid organs/tissues and plasma. TM208 was extracted from the appropriate matrix using methanol followed by centrifugation at 11,255×g for 20min and injection of a 30μL aliquot. Separation was carried out under gradient conditions using an ODS C18 column equipped with a guard column. The mobile phase consisted of methanol and water and retention times of TM208 and plunarizine (IS) were 17.658 and 26.175min, respectively. The analytical procedure provided acceptable precision, accuracy, recoveries and linearity. Stability studies showed that TM208 was stable in organs/tissues homogenates for three freeze–thaw cycles, at room temperature for at least 24h 3weeks at −20°C. The validated method was successfully applied to the determination of TM208 in SD rats following oral administration at a dose of 250mgkg−1.
    Chromatographia 12/2009; 70(11):1721-1725. DOI:10.1365/s10337-009-1402-7 · 1.37 Impact Factor
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    ABSTRACT: 4-Methylpiperazine-1-carbodithioic acid 3-cyano-3,3-diphenylpropyl ester hydrochloride (TM208) is a new compound expected to become a new drug because of excellent in vivo and in vitro anticancer activity and low toxicity. A new, specific and sensitive LC method was set up for detecting the bioavailability of TM208 after oral administration. Samples were extracted with ethyl acetate after oral and intravenous administration. The retention times of TM208 and plunarizine (I.S.) were 5.5 and 9.9min, respectively. The linear range was 0.125–50μgmL−1. The accuracy (error, %) for three concentrations was 2.7–16.6%. Intra-day precision (as RSD) was 1.6–6.9% and inter-day precision was 7.6–11.5%. Extraction recovery of TM208 was 84.15–89.51% and that of the I.S. was 83.3%. Results from stability testing indicated that samples should be analyzed within 24h or frozen immediately for later analysis. The bioavailable fraction (F) calculated by use of a non-compartment model was 63.3%. Pharmacokinetic data for TM208 were: mean residence time 24.3 and 5.1h, V d 186.2 and 35.5Lkg−1, and Cl 6.9 and 4.2Lh−1kg−1 after oral and intravenous administration, respectively. LC–MS comparison of the metabolites after the two methods of administration showed the kind and content of metabolites of TM208 in rat urine after intravenous administration were more than after oral administration. The experimental results show that the low anticancer activity of TM208 after intravenous administration is related to rapid elimination of the drug, and that the kind and content of metabolites do not affect the bioactivity of TM208. KeywordsColumn liquid chromatography-Mass spectrometry-Bioavailability-Metabolites-Dithiocarbamate
    Chromatographia 09/2010; 72(5):459-464. DOI:10.1365/s10337-010-1697-4 · 1.37 Impact Factor
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    ABSTRACT: TM208 and TM209, dithiocarbamate derivatives with potential anti-cancer effects, were evaluated in reversible and time-dependent cytochrome P450 (CYP) 3A inhibition assays in rat liver microsomes using testosterone as probe substrate. Both compounds were found to be weak reversible inhibitors and moderate mechanism-based inhibitors of rat CYP3A. For reversible inhibition on rat CYP3A, the Ki values of competitive inhibition model were 12.10±1.75 and 13.94±1.31 μM, respectively. For time-dependent inhibition, the inactivation constants (Kl) were 31.93±12.64 and 32.91±15.58 μM, respectively, and the maximum inactivation rates (kinact) were 0.03497±0.0069 and 0.07259±0.0172 min−1 respectively. These findings would provide useful in vitro information for future in vivo DDI studies on TM208 or TM209.
    04/2012; 2(2):181–187. DOI:10.1016/j.apsb.2012.02.006