LR12‐peptide quantitation in whole blood by RP‐HPLC and intrinsic fluorescence detection: Validation and pharmacokinetic study

Abstract

A simple, sensitive, selective and robust HPLC method based on intrinsic fluorescence detection was developed for the quantitation of a dodecapeptide (designated as LR12), inhibitor of Triggering Receptor Expressed on Myeloid cells-1, in rat whole blood. Sample treatment was optimized using protein precipitation and solid-phase extraction. Chromatographic separation was carried out in a gradient mode using a core-shell C column (150 × 4.6 mm, 3.6 μm) with mobile phases of acetonitrile and water containing trifluoroacetic acid at 1.0 mL/min. The method was validated using methodology described by the US Food and Drug Administration guidelines for bioanalytical methods. Linearity was demonstrated within the 50-500 ng/mL range and the lower limit of quantitation was 50 ng/mL. Finally, a preliminary pharmacokinetic study after intraperitoneal injection of LR12 in rats was conducted to evaluate both LR12 monomer and its corresponding disulfide dimer, the main product of degradation. Beyond the fact that this paper describes the first fully validated method for LR12 analysis in blood samples, the approach followed here to optimize pre-analytical steps could be beneficial to develop HPLC and/or MS methods for other pharmaceutical peptides.

Simultaneous Determination of Schisandrin and Promethazine with Its Metabolite in Rat Plasma by HPLC-MS/MS and Its Application to a Pharmacokinetic Study

“…Several analytical methods have been reported for the pharmacokinetic determination in biological samples including high-performance liquid chromatography with ultraviolet (UV) detection [14], fluorescence detection (FD) [15], electrochemical detection (ECD) [16], photodiode array (PDA) detection [17], mass spectrometric (MS or MS/MS) detection [18, 19], and some chemiluminescence (CL) technique [20]. However, regarding HPLC-FD, complex sample pretreatment is required to decrease chromatographic interferences from the biological matrix.…”

Triggering receptor expressed on myeloid cells-1 deletion in mice attenuates high-fat diet-induced obesity

“…The TREM-1 KO model can be viewed as a ‘preventive’ model, and then another important question arises: is the modulation of TREM-1 able to reverse an established phenotype? To address this we could use the TREM-1 inhibitory peptide Nangibotide (under clinical study in septic shock patients) but this would require repeated injection as its half-life is very short ( 28 ). The administration of an antagonist antibody would be more practical but it does not exist yet.…”

Pharmacological Autophagy Regulators as Therapeutic Agents for Inflammatory Bowel Diseases

“…Example of this new family of molecules is the peptide LR12 [176] of sequence H-LQEEDTGEYGCV-NH2 that inhibits the triggering receptor expressed on myeloid cells 1 (TREM-1). LR12 was shown to correct the severity of colitis clinically, endoscopically and histologically in a DSS-induced mouse model of colitis [177].…”