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Drug Metabolism and Bioanalysis Letters

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ISSN (Print): 2949-6810
ISSN (Online): 2949-6829

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Research Article

Practical Application of Rodent Transporter Knockout Models to Assess Brain Penetration in Drug Discovery

Author(s): Elin Eneberg*, Christopher R. Jones, Thomas Jensen, Kristine Langthaler and Christoffer Bundgaard

Volume 15, Issue 1, 2022

Published on: 26 April, 2022

Page: [12 - 21] Pages: 10

DOI: 10.2174/1872312815666220222091032

Price: $65

Abstract

Background and Objective: Compound X is a drug candidate for the treatment of neurodegenerative diseases. Its brain distribution was evaluated as part of the lead identification and optimization activities undertaken in early drug discovery.

Methods: The brain distribution of compound X was studied in genetic transporter knockout rodent models, in vivo models with a chemical inhibitor, and in vitro transporter cell systems.

Results: Compound X was found to be a substrate for human Breast Cancer-Resistance Protein (BCRP) in vitro (efflux ratio 8.1) and rodent Bcrp in vivo (Kp, uuKO/Kp, uuWT = 0.15/0.057 = 2.7, p< 0.05) but not a substrate for human P-glycoprotein (P-gp) in vitro (efflux ratio 1.0) nor rodent P-gp in vivo (Kp, uuKO/Kp, uuWT = 0.056/ 0.051 = 1.1, p> 0.05). When both transporters were knocked out in vivo, Kp, uu increased to 0.51±0.02. A similar pattern observed across compounds with related chemistry corroborating the structure-activity relationship.

Conclusion: While in vitro assays showed compound X to be a substrate for human BCRP and not P-gp, in vivo studies indicated a synergistic effect between rodent efflux transporters. However, this only accounted for ~50% of restricted BBB-transport, suggesting involvement of other efflux transporters. Considering Kp, uu as a key criterion for assessing the technical quality of CNS candidates before progression into clinical development, it is important to identify relevant screening assays for a better understanding of low Kp, uu and brain distribution in pre-clinical models for translation to humans.

Keywords: CNS, blood-brain-barrier, ABC transporters, P-gp, BCRP, Kp, uu, drug discovery.

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