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 II project will build upon our successful Phase I demonstration that substrate-free autobioluminescent signal generation can detect both the pre- and post-biotransformed metabolic impacts of therapeutic compounds from a single plate-based assay. Here, we will leverage this technology to develop a panel of industry-relevant autobioluminescent cell lines optimized for the detection of pre- and post-biotransformed compound metabolic impacts and the identification of specific detoxification pathway activation using modern three-dimensional (3D) microphysiological culture systems. These products and their underlying technology will specifically 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. By optimizing this technology to function within the industry-preferred 3D microphysiological format, we will address the critical need for new methods that can both identify compound toxicity and elucidate the mechanisms through which cells mitigate the compounds? effects. The autonomous nature of this technology will increase toxicological data acquisition while preserving the critical advantage of presenting physiologically- relevant data, and reducing the cost of performance by eliminating substrates, reducing complexity, limiting hands-on operation time, obviating the need for sample destruction, and reducing the potential for measurement error. Through the validation of this technology at a scale relevant to tier 1 drug discovery screening and its comparative analysis against the existing gold-standard ATP content assay, this revolutionary approach is poised to have a significant and immediate impact towards reducing the estimated $8B/year in unnecessary expenditures made by pharmaceutical companies during their development of the 48% of new compounds that fail at the Phase I clinical trial stage due to misidentification of toxicological effects during tier 1 screening. This is possible because, as demonstrated in our Phase I work, the use of our autobioluminescent technology overcomes the high economic and logistical costs of existing, traditionally-bioluminescent cell?s requisite chemical substrate addition, which must co-occur with each generation of signal, and the intensive hands-on time necessitated to scale cultures due to their requisite sample destruction concurrent with imaging. Similarly, our autobioluminescent technology also obviates the hurdles presented by fluorescent cell?s susceptibility to autofluorescent signal inhibition and their tendency to remain active during downturns in cellular metabolism or even after cell death. The technology and products developed in this effort will therefore be capable of significantly improving the throughput and effectiveness of microphysiological systems-based tier 1 compound screening to improve the efficiency and economics of new compound development, and ultimately, consumer safety. This will allow them to thrive in a microphysiological system market that is predicted to maintain a compound annual growth rate of 70% to exceed $1.3B globally by 2022.
Failure to identify the toxicity of pharmaceutical compounds such as Merck?s Vioxx, Bayer?s Baycol, and Wyeth?s FenPhen have caused consumers to be injured or killed by the drugs that were supposed to help them, and resulted in negative publicity and legal fees costing billions of dollars for the companies that produced them. While pharmaceutical companies have significantly enhanced their early stage toxicity screening regimens by transitioning to complex cell culture systems that can better identify new compounds? toxicological effects on the human body to avoid repeating these mistakes, the use of these systems has increased the cost of new drug development to a level that is unsustainable for both consumers and the companies themselves due to a lack of technologies capable of economically identifying toxicity in these modern systems. In this Phase II R&D effort, 490 BioTech proposes to implement a novel toxicological screening technology based upon a synthetic luciferase genetic construct that links cellular health to the autonomous production of light in the visible spectrum to improve toxicity data acquisition by increasing the throughput and reducing the cost of modern pharmacological development assays while preserving their increased level of safety.
| Xu, Tingting; Conway, Michael; Frank, Ashley et al. (2017) Co-Cultured Continuously Bioluminescent Human Cells as Bioreporters for the Detection of Prodrug Therapeutic Impact Pre- and Post-Metabolism. Sensors (Basel) 17: |
| Xu, Tingting; Close, Dan; Handagama, Winode et al. (2016) The Expanding Toolbox of In Vivo Bioluminescent Imaging. Front Oncol 6:150 |
