RT Journal Article
SR Electronic
T1 Use of Cryopreserved Hepatocytes as Part of an Integrated Strategy to Characterize In Vivo Clearance for Peptide-Antibody Conjugate Inhibitors of Nav1.7 in Preclinical Species
JF Drug Metabolism and Disposition
JO Drug Metab Dispos
FD American Society for Pharmacology and Experimental Therapeutics
SP dmd.119.087742
DO 10.1124/dmd.119.087742
A1 Robert S Foti
A1 Kaustav Biswas
A1 Jennifer Aral
A1 Xuhai Be
A1 Loren Berry
A1 Yuan Cheng
A1 Kip Conner
A1 James R Falsey
A1 Charles Glaus
A1 Brad Herberich
A1 Dean Hickman
A1 Tayo Ikotun
A1 Hongyan Li
A1 Jason Long
A1 Liyue Huang
A1 Les P Miranda
A1 Justin Murray
A1 Bryan Moyer
A1 Chawita Netirojjanakul
A1 Thomas E Nixey
A1 Kelvin Sham
A1 Marcus Soto
A1 Christopher M Tegley
A1 Linh Tran
A1 Bin Wu
A1 Lin Yin
A1 Dan A Rock
YR 2019
UL http://dmd.aspetjournals.org/content/early/2019/08/06/dmd.119.087742.1.abstract
AB The identification of non-opioid alternatives to treat chronic pain has received a great deal of interest in recent years. Recently, the engineering of a series of Nav1.7 inhibitory peptide-antibody conjugates has been reported and herein, the preclinical efforts to identify novel approaches to characterize the pharmacokinetic properties of the peptide conjugates are described. A cryopreserved plated mouse hepatocyte assay was designed to measure the depletion of the peptide-antibody conjugates from the media, with a correlation being observed between percent remaining in the media and in vivo clearance (Pearson r = -0.5525). Physicochemical (charge and hydrophobicity), receptor-binding (FcRn) and in vivo pharmacokinetic data were generated and compared to the results from an in vitro our hepatocyte assay that was hypothesized to encompass all of the aforementioned properties. Correlations were observed between hydrophobicity, FcRn binding, depletion rates from the hepatocyte assay and ultimately, in vivo clearance. Subsequent studies identified potential roles for the low-density lipoprotein and mannose/galactose receptors in the association of the Nav1.7 peptide conjugates with mouse hepatocytes, though in vivo studies suggested that FcRn was still the primary receptor involved in determining the pharmacokinetics of the peptide conjugates. Ultimately, the use of the cryopreserved hepatocyte assay along with FcRn binding and hydrophobic interaction chromatography provided an efficient and integrated approach to rapidly triage molecules for advancement while reducing the number of in vivo pharmacokinetic studies.SIGNIFICANCE STATEMENT While multiple in vitro and in silico tools are available in small molecule drug discovery, pharmacokinetic characterization of protein therapeutics is still highly dependent upon the use of in vivo studies in preclinical species. The current work demonstrates the combined use of cryopreserved mouse hepatocytes, hydrophobic interaction chromatography and FcRn binding to characterize a series of Nav1.7 peptide-antibody conjugates prior to conducting in vivo studies, thus providing a means to rapidly evaluate novel protein therapeutic platforms while concomitantly reducing the number of in vivo studies conducted in preclinical species.