Development of a high-throughput screen to detect the effects of both pre- and post-biotransformed compounds for enhanced content drug discovery workflows

DESCRIPTION provided by applicant Development of a high throughput screen to detect the effects of both pre and post biotransformed compounds for enhanced content drug discovery workflows Project Summary This Small Business Innovation Research Phase I project proposes to engineer a panel of autonomously bioluminescent human cell lines for the simultaneous high throughput detection of both pre and post biotransformed cytotoxicity onsets resulting from drug treatment across multiple tissue types to address the National Institute of General Medical Sciences NIGMS request for novel in vivo and in vitro methods for predicting the safety and toxicities of pharmacologic agents With an average of novel molecules that must be screened for each new lead compound developed and an average of to years of research and development at a cost of up to $ B to manufacture one new drug pharmaceutical companies must develop new testing regimens that provide more data at a lower cost in order to achieve the economics necessary to remain profitable Up to of failures for these new compounds at the clinical level are related to cytotoxicity which often onl manifests during the costly and time consuming process of whole animal testing This problem could be significantly mitigated by coordinately screening multiple tissue types simultaneously in a high throughput fashion during upstream tier screening However with existing bioluminescent reporter technology this is simply not possible because the current market of bioluminescent reporter cells being applied toward toxicology screening relies upon a firefly luciferase gene construct that must be provided with a chemical substrate to activate its light emission response resulting in only marginally informative single time point snapshots of potential toxicological interactions In contrast our substrate free autobioluminescent reporter cell lines emit light continuously and can modulate the output level of this signal in real time in response to cytotoxic interactions This provides an uninterrupted stream of visual data over the lifetime of the reporter cell as it interacts and reacts to compound exposure at both its pre and post biotransformed states Furthermore by leveraging human cells as the sensing platform our assay provides more accurate and realistic information in regards to bioavailability and a chemicalandapos s true effect on individual human health than does the employment of small animal models With current in vitro screening assays now representing a $ B market with a predicted annual growth rate we believe we possess a product capable of significantly impacting the chemical drug screening market and here in particular advancing our understanding of cytotoxic chemical biotransformations as they pertain to public health and consumer safety PUBLIC HEALTH RELEVANCE The drug discovery process requires that upwards of molecules be screened for each new lead compound developed and relies upon an expensive and time consuming combination of in vitro cell culture based and in vivo whole animal based models to identify validate and ensure the safety of any resulting potential therapeutic agents This process is frustratingly exacerbated by an unfortunate dichotomy whereby the inexpensive in vitro cell culture systems used for tier screening contribute to failures due to their inabiliy to model the complexity and parallel systems interaction inherent in whole animal models which are responsible for up to of new compound failures at the clinical level and because they are not capable of demonstrating species specific effects To overcome these detractions and develop an improved tier screening system that reduces the cost and time required for new compound evaluation BioTech proposes to develop a panel of multiple continuously bioluminescent human cell lines that will permit the simultaneous monitoring of each line to ascertain both the individual effects of compound treatment as well as the downstream effects of a compoundandapos s biotransformed metabolic breakdown products in real time