Toxins from venomous animals are extremely potent and selective towards defined targets involved in numerous physiological processes, such as ion channels and receptors. This work aims to develop innovative approaches to elucidate and characterize molecules issued from venoms, potentially exploitable for pharmacological, cosmetics and medical applications, with a particular focus on spider venoms. A structure-driven strategy, mainly based on transcriptomics and proteomics techniques, was developed and allowed uncovering the peptide toxins arsenal of four unstudied spider species. A bioactivity-guided strategy was used to highlight acylpolyamines from a spider venom as quinone reductase inducers. Finally, a biocomputing-assisted strategy was designed and applied to the study of peptides, leading to the discovery and characterization of novel antifungal agents.