New strategies leveraging ALIAmides in acute and chronic anti-inflammatory therapies

This thesis examines innovative approaches to enhance the anti-inflammatory and antiproliferative potential of ALIAmides, bioactive lipids with promising roles in modulating physiological and pathological inflammatory responses. ALIAmides, particularly Palmitoylethanolamide (PEA) and novel compounds like N-palmitoyl-D- glucosamine (PGA) and adelmidrol, exert significant anti-inflammatory effects primarily through PPAR-α activation, which suppresses pro-inflammatory cytokines and enhances anti-inflammatory and antioxidant gene expression. However, pharmacokinetic limitations, including poor bioavailability, restrict their therapeutic application. This research addresses these challenges by investigating novel delivery systems and synthetic alternatives in murine models of inflammatory bowel disease (IBD), colorectal cancer (CRC), and SARS-CoV-2-induced lung inflammation. To optimize ALIAmide administration, micronized PGA (m-PGA) and adelmidrol were formulated to improve absorption and efficacy, with m-PGA tested in a DNBS (dinitrobenzene sulfonic acid)-induced colitis model to evaluate its impact on inflammation during both acute and resolution phases. In addition, m- PGA was investigated for its antiproliferative properties in a colorectal cancer model induced by AOM + DSS (azoxymethane + dextran sodium sulfate), focusing on its effects on inflammatory markers, tumor size, and cellular proliferation. To further enhance targeted delivery, an engineered strain of Lactobacillus paracasei F19 (pNAPE-LP) was developed, capable of biosynthesizing PEA at the site of inflammation in response to palmitate supplementation, with its efficacy assessed in a murine model of SARS-CoV-2-induced lung inflammation. A novel adelmidrol- hyaluronic acid rectal gel was also evaluated in a murine colitis model and ex vivo in human biopsy samples for its potential to reduce inflammation and protect mucosal integrity. The m-PGA demonstrated significant anti-inflammatory effects in the colitis model, reducing cytokine expression and improving histological scores in both acute and resolution phases. In the CRC model, m-PGA inhibited tumor proliferation, reduced inflammation, and showed marked improvements in tissue integrity. The adelmidrol-hyaluronic acid gel (Ade/HA) produced similar anti- inflammatory effects, effectively reducing pro-inflammatory markers and preserving mucosal integrity. Notably, pNAPE-LP in the lung inflammation model showed 8 promising results in mitigating inflammatory cell infiltration, preserving alveolar structure, and downregulating ACE-2 expression, indicating its potential as an adjunctive therapy for SARS-CoV-2-related respiratory distress. The results underscore the potential of ALIAmides as versatile agents for managing inflammation through both oral and topical formulations. The findings support the hypothesis that m-PGA and adelmidrol can provide effective anti- inflammatory and antiproliferative benefits in IBD and CRC by modulating PPAR-α pathways, offering therapeutic advantages with fewer side effects than traditional treatments. The engineered pNAPE-LP strain represents a groundbreaking approach to in situ drug delivery, capable of enhancing bioavailability while minimizing systemic exposure. This research suggests exciting directions for optimizing ALIAmide-based therapies through pharmaceutical and biotechnological innovations and contributes to advancing anti-inflammatory strategies, positioning ALIAmides as promising candidates for novel therapies in chronic inflammation and cancer.