SBIR-STTR Award

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase I project is to develop technology using engineered bacteria to improve protein stability in two large markets: therapeutic proteins and industrial enzymes. Proteins used as therapeutics frequently have insufficient half-lives in human blood plasma, so that patients with chronic disease need to receive frequent dosings, often by painful injections. These burdensome dosing schedules result in high rates of patient noncompliance, with attendant poor responses and negative health outcomes. Therapeutic proteins with improved half-lives in blood plasma would permit relaxed dosing schedules, lowering costs of administration and improving outcomes by reducing noncompliance. This proposed technology focuses on improving the plasma half-life of a therapeutic for a chronic disease primarily affecting children that currently comprises a $4B global market. Similarly, industrial enzymes are frequently deployed in harsh environments that impair enzyme stability and activity. Endowing enzymes with improved resistance to destabilizing chemicals would enable their deployment in high-value environments that are currently prohibitive. The proposed technology will be used to stabilize an enzyme for application in a $730M segment of the personalized medicine market. If successful, this work would provide a platform for developing next-generation enzymes deployable in currently prohibitive environments. The intellectual merit of this SBIR Phase I project is to utilize a synthetic biology platform to create proteins with new amino acid building blocks that dramatically improve half-life and stability. Many proteins used as therapeutics or used as industrial enzymes are stabilized by disulfide bonds. These bonds break in the presence of chemicals called reducing agents that are found both in human blood plasma (destabilizing to therapeutics) and in solvents and buffers (destabilizing to industrial enzymes). Using a strain of engineered E. coli that can incorporate amino acids beyond the 20 standard amino acids into proteins, the goal is to replace disulfide-forming cysteine amino acids with selenocysteine amino acids that form bonds called diselenides. Diselenide-stabilized proteins maintain stability and activity in environments with reducing agents incompatible with disulfide-stabilized proteins. This project will produce a diselenide-stabilized protein therapeutic for a major disease class. Improvements to disulfide-stabilized therapeutics will be demonstrated by ELISA assays showing improved binding activity, and by cell-based assays showing improved therapeutic activity, after prolonged exposure to blood plasma. This project also will produce a diselenide-stabilized industrial enzyme to catalyze a critical reaction for personalized medicine. Performance improvements over disulfide-stabilized enzymes will be demonstrated by in vitro stability and activity assays in reducing conditions.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

The broader impact of this Small Business Innovation Research (SBIR) Phase II project is to help patients living with diabetes. The disease accounts for 12% of deaths in the US and patients face major lifestyle changes. Most patients transition to insulin replacement therapy, which carries a complex dosing schedule that, if not followed closely, can leave patients in dangerous states of glucose dysregulation. More than 50 million diabetics currently use basal insulin analogs designed for longer activity than human insulin. The convenience and improved safety of these analogs has led to widespread adoption and a global market surpassing $10B. However, all current basal insulins require daily injections, a dosing burden that leads to poor treatment adherence, leaving patients vulnerable to dangerous fluctuations in blood glucose. The modified insulin described in this Phase II project is intended to provide the stability necessary to achieve once-weekly dosing. Relaxing the injection schedule should dramatically improve compliance and safety for patients; furthermore, the solution can be delivered at lower cost. The project uses a scalable in vivo protein production platform to produce long-acting insulin analogs for the diabetes market. The project utilizes the platform?s unique capability to site-specifically install non-standard amino acids into proteins, and to produce the modified proteins at scale. By replacing key bond-forming amino acids in insulin with non-standard amino acids that form stronger bonds, the modified insulins can achieve the stability necessary to support once-weekly dosing. The research objectives are to: produce sufficient quantities of variants of this insulin analog to support an experimental program, demonstrate improved stability of the variants over wild-type insulin in cell-based assays, and demonstrate sufficiently prolongated pharmacodynamics of the insulin analogs in an animal study to support once-weekly dosing. Potential outcomes include the first insulin analog capable of filling a major clinical and commercial need for affordable, safe insulin analogs with relaxed dosing schedules. Further, the work provides technical validation of a novel protein production platform.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.