Validation of Immunometabolic NLRX1 Therapeutics for IBD

Validation of Immunometabolic NLRX1 Therapeutics for IBD Biotherapeutics Inc (BTI) is an emerging biotech company that synergistically combines the power of advanced computational modeling with translational experimentation to accelerate the development of novel products for precision medicine and health. This SBIR application stems from data showing a vital role for nucleotide-binding oligomerization domain, leucine rich repeat containing X1 (NLRX1) as a new therapeutic target for IBD. Our Product: BTI has identified the first family of small-molecule compounds that bind and activate the novel regulatory molecule, NLRX1. In a Phase I SBIR, we successfully established NX-13 as a NLRX1-specific agonist that reduces inflammatory lesions by 90% in CD4+ T cell-specific immunometabolic mechanisms. Background: IBD is a chronic widespread and debilitating illness that afflicts over 5 million people worldwide with total expenses exceeding $15 billion annually in the U.S alone. Current treatments are only modestly successful with significant adverse side effects. Thus, there is an unmet clinical need for safer, more efficacious IBD therapeutics. NLRX1 can suppress intestinal inflammation during infections and autoimmune disorders. BTI has validated that loss of NLRX1 causes reduced mucosal healing, increased fibrosis, andgt;5-fold up-regulation in inflammatory cytokine biomarkers, and complete restructuring of gut microbiome ecology during IBD. This SBIR Phase II application will characterize microbial and anti-fibrotic mechanisms of NX-13, demonstrate translational efficacy in human UC primary cells and conduct IND-enabling safety studies. The Specific Aims are to: AIM 1. Determine the effect of NX-13 on the intestinal microbiome during colitis through 16S gut microbiome sequencing and signatures of gut-microbiome interactions. AIM 2. Characterize the anti-fibrotic mechanisms of NX-13 in chronic colitis models and in vitro effects on fibroblast gene expression and extracellular matrix deposition. AIM 3. Validate the translational efficacy of NX-13 in PBMCs and LPMCs from UC patients through evaluation of overall cellular response and CD4-specific immunometabolic mechanisms. AIM 4. Conduct an IND-enabling GLP toxicity study in a non-rodent species with a 28-day repeat dose toxicity study at three dose levels (250, 500, 1000 mg/kg) compared to vehicle. Expected Outcomes: Validation of NX-13 as a lead agonistic molecule for targeting NLRX1 through: i) identification of gut microbial and anti-fibrotic mechanisms of NX-13; ii) 50% reduction in TNFα and IFNγ production in human PBMCs and LPMCs; and iii) a benign safety profile with oral NOAEL ≥ 1,000 mg/kg. Commercial Application: Success in this project will launch a new drug development pipeline centered on NLRX1-activating therapeutics with anti-inflammatory and anti-fibrotic effects. BTI’s new NLRX1-targeting oral therapeutics could disrupt a market of over $10B annually growing at 25% annual rates.Public Health Relevance The spread of Inflammatory bowel disease (IBD) is growing globally and it afflicts over 2 million people in the United States and over 5 million people worldwide. This SBIR Phase I has the potential to develop a novel class of potent oral drugs for IBD that exert their therapeutic actions by acting on the novel target, NLRX1, and advance them along the FDA regulatory pipeline into IND and clinical testing. The proposed studies will lay the groundwork for developing NX-13 a small molecule therapeutic with the potential to selectively engage and activate NLRX1 in the gastrointestinal (GI) tract and discovering how NX-13 modulates immune and inflammatory responses in the GI tract in IBD.