Toll-like receptors (TLRs) are transmembrane proteins that play pivotal roles in immune defence system in different kinds of organisms. They recognise pathogen associated molecular patterns (PAMPs) and danger associated molecular patterns (DAMPs) and the recognition leads to quick responses to eliminate the pathogen from the host. Ligand stimulation of TLR triggers the activation of their cytosolic Toll/interleukin-1 receptor (TIR) domains, which can recruit other intracellular, TIR-containing adaptor proteins, including myeloid differentiation primary response gene 88 (MyD88) and MyD88 adaptor-like (MAL) via TIR-TIR interactions. The stimulation of cell surface TLRs results in the cytosolic TIR domains from at least two TLRs to come into proximity and form a scaffold for downstream signalling. The adapter protein MAL has been studied extensively and is a key player in TLR signalling cascade of human immune system. The cytosolic MAL protein is recruited upon activation of TLR4 and TLR5, or the combination of TLR2 with either TLR1 or 6. Upon activation MAL forms filaments, which further leads to the formation of the functional TLR signalosomes and subsequent NF-κB activation. It was reported in mice with the disease-associated MAL polymorphism D96N are protected from lipopolysaccharide induced lethality in-vivo and have reduced inflammation, validating the therapeutic potential of MAL in the TLR4 signalosome. Hence, MAL represents a promising target for inhibition with broad therapeutic applications. Previous studies have identified o-vanillin to disrupt higher order assembly formation of MALTIR filament and there are no other small molecule inhibitors reported so far for MALTIR. Previous study from the lab, identified few small molecule binders using 19F fragments-based drug discovery (FBDD) pipeline with few positive hits showing binding using both NMR and SPR. Here, we will extend the characterisation of the interactions between the shortlisted molecules against MALTIR and will analyse the inhibitory activity of the molecules on MALTIR filament formation using turbidity assay. The integrated data will provide the basis for establishing the structure-activity relationship of these interactions and allow us to chemically elaborate the active hits to achieve higher potency. The project aims to develop lead therapeutic agents for treating the debilitating inflammatory diseases.