Pharmacokinetic Characterization of the DDAH1 Inhibitors ZST316 and ZST152 in Mice Using a HPLC-MS/MS Method

Abstract

The pharmacokinetic profile of ZST316 and ZST152, arginine analogues with inhibitory activity towards human dimethylarginine dimethylaminohydrolase-1 (DDAH1), was investigated in mice using a newly developed HPLC-MS/MS method. The method proved to be reproducible, precise, and accurate for the measurement of the compounds in plasma and urine. Four-week-old female FVB mice received a single dose of ZST316 and ZST152 by intravenous bolus (30 mg/Kg) and oral gavage (60 mg/Kg). ZST316 Cmax was 67.4 µg/mL (intravenous) and 1.02 µg/mL (oral), with a half-life of 6 h and bioavailability of 4.7%. ZST152 Cmax was 24.9 µg/mL (intravenous) and 1.65 µg/mL (oral), with a half-life of 1.2 h and bioavailability of 33.3%. Urinary excretion of ZST152 and ZST316 was 12.5%–22.2% and 2.3%–7.5%, respectively. At least eight urinary metabolites were identified. After chronic intraperitoneal treatment with the more potent DDAH1 inhibitor, ZST316 (30 mg/Kg/day for three weeks), the bioavailability was 59% and no accumulation was observed. Treatment was well tolerated with no changes in body weight vs. untreated animals and no clinical signs of toxicity or distress. The results of this study show that ZST316 has a favorable pharmacokinetic profile, following intraperitoneal administration, to investigate the effects of DDAH1 inhibition in mice.

Development of a HPLC-MS/MS Method to Assess the Pharmacokinetics and Tumour Distribution of the Dimethylarginine Dimethylaminohydrolase 1 Inhibitors ZST316 and L-257 in a Xenograft Model of Triple-Negative Breast Cancer in Mice

“…The pharmacokinetics and safety profile of the most promising DDAH1 inhibitors in our series, compounds ZST316 and ZST152, have been recently investigated in fourweek-old Friend leukemia virus B (FVB) mice after oral, intravenous, and intraperitoneal administration. In this study, the most potent DDAH1 inhibitor, ZST316 (Ki: 1 µM [22], subsequently revised to 261 nM using a more performant assay [30], personal data) exhibited a favourable pharmacokinetic profile and excellent tolerability [31]. Notably, further analyses also led to the identification of several urinary metabolites of ZST316, including the known DDAH1 inhibitor, compound L-257 [16,18,32].…”

“…A quantitation method for ZST316 was previously established [31]. Here, we set up, validated, and applied the method for the combined measurement of ZST316 and L-257.…”

“…Lower concentrations (i.e., 1.5 ng/mL) were indistinguishable from the background noise (S/N < 3). We established a LOQ of 5 ng/mL for L-257, in line with that of ZST316 [31], with precision of 5.2% and accuracy of 107.8% (n = 5). The linearity of the method for L-257 was acceptable over the concentration range 5.0-1000.0 ng/mL, with a coefficient of determination (R 2 ) of 0.996.…”

Recent advances in DDAH1 inhibitor design and discovery: insights from structure–activity relationships and X-ray crystal structures

“…26,28,32 Also for most of the DDAH1 inhibitors (both arginine and non-arginine-like) limited information of their pharmacokinetic proles are available with only a few studies performed in animal models. Pharmacokinetic data is available for L-257 (13), ZST316 (20) and ZST152 (24) 40,41 and animal studies detailing the effects of DDAH1 inhibition of L-257 (ref. 34) and DD1E5.…”

“…A recent study conducted by our research group 41 assessed the pharmacokinetics of 20 and 24 ( ZST316 and ZST152 respectively, Fig. 3), inhibitors of h DDAH1.…”