Drug metabolism and pharmacokinetic testing (DMPK) is crucial for understanding the clearance mechanisms, clearance rate, and any potential drug-drug interactions of small molecule drugs during lead optimization prior to initiating in-human clinical trials. DMPK testing is currently performed using animal models, usually rats. However, these studies are problematic for several reasons: 1) Animal-based DMPK studies are expensive and create a large financial burden during lead optimization; the top 20 pharmaceutical companies spend >$1.2 billion each year on PK testing, with over 1 million animals used by industry per year; 2) They are time-consuming and delay the collection of important data. For example, the synthesis of each compound needs to be scaled up from µg to mg quantities before rat studies can be initiated. 3) While higher animal species, such as dogs, pigs, and non-human primates, have greater human relevance, they are more expensive than rodent studies and require synthesis of even greater quantities of active pharmaceutical ingredient (API); and 4) The ability of animal models to predict human outcomes is controversial. Although recent attempts have been made to identify in vitro approaches, these are all limited in terms of reliability, long-term analysis ability, reproducibility, poor reflection of in vivo hepatic transport, clearance, and metabolism. Thus, the pharmaceutical industry needs better approaches for DMPK testing that are more predictive, cheaper, and faster than currently available in vitro and in vivo methods. Successful development of such a system will improve safety and help reduce the ~90% of drug candidates that currently fail in clinical trials. In solution to this unmet need, we propose an integrated (in vitro and in silico) hepatic clearance platform that merges a human liver tissue chip and translational software. This technology can predict human hepatic clearance parameters accurately in 2 weeks without the need of API scale-up. Javelin is the only organization pursuing this technology for drug metabolism & disposition, which requires design features that are unique to DMPK studies and that cannot be met using microphysiological systems designed for toxicology and pharmacology applications. The purpose of this Direct to Phase II SBIR proposal is improve our technology ready for launch by optimizing the chip design to enhance usability, evaluate drug clearance mechanisms, and assess the clinical translation of our technology. This will be achieved through the following Aims: 1) System design optimization; 2) Evaluation and characterization of known drug clearance mechanisms on the Javelin hepatic clearance chip; and 3) Assessment of in vitro to in vivo translation to predict clinical outcomes. Successful application of our tissue chip perfusion system will reduce the need for other preclinical models to estimate drug PK outcomes, thereby reducing the risk and cost associated with drug development. Indeed, ou...