Quantitation of a Novel Engineered Anti-infective Host Defense Peptide, ARV-1502: Pharmacokinetic Study of Different Doses in Rats and Dogs

Abstract

The designer proline-rich antimicrobial peptide (PrAMP) Chex1-Arg20 amide (ARV-1502) is active against Gram-negative and Gram-positive pathogens in different murine infection models when administered parenterally and possesses a wide therapeutic index. Here we studied the pharmacokinetics of ARV-1502 for the first time when administered intramuscularly or intravenously (IV) in Sprague Dawley rats and Beagle dogs. First, a specific and robust quantitation method relying on parallel reaction monitoring (PRM) using a high-resolution hybrid quadrupole-Orbitrap mass spectrometer coupled on-line to reversed-phase uHPLC was established and validated. The limit of detection was 2 ng/mL and the limit of quantitation was 4 ng/mL when spiked to pooled rat and dog plasma. When ARV-1502 was administered IV at doses of 75 and 250 μg/kg in dogs and rats, the plasma concentrations were 0.7 and 3.4 μg/mL 2 min post-administration, respectively. ARV-1502 plasma concentrations declined exponentially reaching levels between 2 and 4 ng/mL after 2 h. Intramuscular administration of 0.75 mg/kg in dogs and 2.5 mg/kg in rats resulted in a different pharmacokinetics profile. The plasma concentrations peaked at 15 min post-injection at 1 μg/mL (dogs) and 12 μg/mL (rats) and decreased exponentially within 3 h to 4 and 16 ng/mL, respectively. The initial plasma concentrations of ARV-1502 and the decay timing afterwards indicated that the peptide circulated in the blood stream for several hours, at some point above the minimal inhibitory concentration against multidrug-resistant Enterobacteriaceae, with blood concentrations sufficient to suppress bacterial growth and to modulate the immune system.

In Vitro Properties and Pharmacokinetics of Temporarily PEGylated Onc72 Prodrugs

“…This also leads to a fast clearance of Onc72 with an elimination t 1/2 of only ≈14 min when administered intraperitoneal in mice, although Onc72 levels in organs were relatively stable. [ 10 ] Similar observations were reported for Api137 in mice and ARV‐1502 in rats and dogs, [ 9,34 ] which requires higher doses with the risk of adverse effects, multiple dosing, or continuous delivery strategies. [ 28 ] These limitations can be overcome for many peptides by improving their binding to a plasma protein leading to low steady‐state concentrations of the free peptide, or by covalently coupling them to a large polymer or biopolymer, such as PEG.…”

“…LC-MS: Sample preparation relied on Oasis HLB 96-well SPE plates using a slightly modified protocol previously reported. [34] The stationary phase was conditioned with acetonitrile (500 μL), washed with aqueous acetonitrile (40%, v/v; 500 μL) containing formic acid (0.1%, v/v), and equilibrated with aqueous TFA (0.1%, v/v; 500 μL). Aliquots of plasma or urine (10 or 1 μL) were spiked with isotope-labeled Onc72 and [1][2][3][4][5][6][7][8][9][10][11] Onc72 as internal standard (62.5 or 125 fmol μL −1 ).…”

Functional Effects of ARV-1502 Analogs Against Bacterial Hsp70 and Implications for Antimicrobial Activity

“…Peptide Chex1-Arg20 (recently termed ARV-1502; Chex-RPDKPRPYLPRPRPPRPVR-NH 2 ; Chex denotes 1-amino-cyclohexane carboxylic acid) and its dimer A3-APO were designed based on a sequence comparison of different naturally occurring PrAMPs, such as pyrrhocoricin and drosocin ( Otvos et al, 2005 ; Noto et al, 2008 ), to obtain analogs with improved preclinical properties against various infections. In diverse studies, ranging from MICs to infection models and finally pharmacokinetics, ARV-1502 proved to be a very promising candidate against infections induced by Enterobacteriaceae ( Ostorhazi et al, 2017 ; Otvos et al, 2018 ; Brakel et al, 2019 ). Its binding to the bacterial chaperone DnaK, which was first identified as the lethal target but is currently considered as a secondary, non-lethal, target after the 70S ribosome, was thoroughly investigated ( Zahn et al, 2013 ).…”

Strategies in Translating the Therapeutic Potentials of Host Defense Peptides

“…This study suggested that rational insertion of Pro residue through SAR analysis could improve the biological membrane selectivity of microbicidal peptides. Proline-rich designed HDPs such as ARV-1502 (42), oncocin (43), and Bac-5 (44,45) have shown significant efficacy against Gram-negative pathogens but not host cells membranes. Unlike cationic HDPs, proline-rich peptides kill bacteria through inhibition of protein synthesis (12,(45)(46)(47).…”