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
| Autor: | Brad Herberich, Thomas Nixey, Lin Yin, Tayo Ikotun, Kelvin K. C. Sham, Jason Long, Hongyan Li, Charles Glaus, Xuhai Be, Les P. Miranda, Yuan Cheng, Marcus Soto, James R. Falsey, Bryan D. Moyer, Robert S. Foti, Christopher M. Tegley, Chawita Netirojjanakul, Linh Tran, Kip P. Conner, Jennifer Aral, Liyue Huang, Kaustav Biswas, Justin K. Murray, Bin Wu, Loren Berry, Dean Hickman, Dan A. Rock |
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| Rok vydání: | 2019 |
| Předmět: |
Male
Immunoconjugates Metabolic Clearance Rate In silico Drug Evaluation Preclinical Pharmaceutical Science Peptide Receptors Fc Mice Neonatal Fc receptor In vivo medicine Animals Tissue Distribution Receptor Cryopreservation Mice Knockout Voltage-Gated Sodium Channel Blockers Pharmacology chemistry.chemical_classification Drug discovery Histocompatibility Antigens Class I NAV1.7 Voltage-Gated Sodium Channel Antibodies Monoclonal In vitro Macaca fascicularis medicine.anatomical_structure Biochemistry chemistry Hepatocyte Hepatocytes Administration Intravenous Chronic Pain Peptides |
| Zdroj: | Drug Metabolism and Disposition. 47:1111-1121 |
| ISSN: | 1521-009X 0090-9556 |
| Popis: | The identification of nonopioid 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 percentage remaining in the media and in vivo clearance (Pearson r = -0.5525). Physicochemical (charge and hydrophobicity), receptor-binding [neonatal Fc receptor (FcRn)], and in vivo pharmacokinetic data were generated and compared with the results from our in vitro hepatocyte assay, which was hypothesized to encompass all of the aforementioned properties. Correlations were observed among 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, although 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: Although 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 hepatocytes, hydrophobic interaction chromatography, and neonatal Fc receptor 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. |
| Databáze: | OpenAIRE |
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