Targeting mitochondria by ceramide in head and neck cancers - SUMMARY Introduction: Lipo-Immuno Tech (LIT), LLC, is a small biotech company that originated from the Hollings Cancer Center (Medical University of South Carolina), which focuses on cancer therapeutics. This Fast-track R42 application aims to develop a potent, specific, and well-tolerated C18-ceramide analog drug that selectively induces lethal mitophagy to inhibit tumor growth and enhance immunotherapy in head and neck squamous cell carcinoma (HNSCC). Significance: HNSCC represents over 71,000 new cancer cases, with about 16,000 cancer-related deaths in the United States annually. There is an unmet clinical need for the development of novel therapeutic agents to treat this disease and enhance response to immunotherapy in advanced HNSCC. Innovation and the Product: Lower C18-ceramide levels are associated with advanced disease stages and poor survival in patients with HNSCC. To selectively restore C18-ceramide accumulation in HNSCC mitochondria, we developed a first-in-class novel lead compound, LCL768, which conjugates C18- ceramide with pyridinium, targeting C18-ceramide to HNSCC mitochondria to bind LC3B and recruit autophagosomes (IC50 = 100 nM). Additionally, selenite is included to induce Drp1 activation and lethal mitophagy, leading to tumor suppression. The patent application for LCL768 and its derivatives has recently been issued (US 12,390,490 B2, August 19, 2025). Preliminary Data: Based on our initial optimization studies of 30 ceramide analogs, we selected LCL768 as the lead compound, which also reduced the number of exhausted CD8+ T cells and M2 macrophages in the HNSCC tumor microenvironment. We will complete our Fast-Track STTR-R42 application with three Specific Aims: Phase I-Specific Aim: SAR and optimization studies to improve the on-target potency of the lead LCL768 and its backup analog drug. Milestones and benchmarks of success (Stop/Go Determinations): Upon completion of Phase I, we expect to have LCL768 derivatives with improved on-target potency, specificity, and selectivity towards LC3B binding and HNSCC mitochondria accumulation, as well as strong Drp1 activation and mitophagy induction with fumarate depletion (a biomarker for mitochondrial metabolism inhibition). Phase II- Specific Aim 1: Lead optimization to test drug- likeness and ADME (absorption, distribution, metabolism, and excretion) properties of LCL768 and its backup derivative. Specific Aim 2. PK/PD and toxicity studies, as well as testing the lead LCL768 and its backup derivative, to enhance response and overcome resistance to immunotherapy in vivo. Milestones and benchmarks of success: A lead LCL768 or its selected analog drug, showing bioavailability of >20-30% (F > 20-30%) in two species and in vivo tumor suppressor activity and immunotherapy sensitization (TGI > 80%), without toxicity, will be considered an optimized lead for an IND application.
