Two hydroxy-cinnamic acid derivatives (I and II) were synthetised by the reaction of Caffeic acid, m-hydroxybenzoic acid with p-aminomethylbenzoic acid (AMBA), respectively. The chemical structures were confirmed by MS, IR, 1H-NMR and 13C-NMR. The interactions between compounds and HSA were investigated under imitated physiological condition by fluorescence spectroscopy, UV-visible absorption spectroscopy and Molecular docking. The experimental results indicated that the quenching mechanism of HSA by the derivatives was static quenching mechanism at different temperatures. The derivatives-HSA interactions were mainly driven by hydrogen bonds and van der Waals’ forces. Meanwhile, derivatives I and II affected the conformation of HSA. The molecular docking represented that derivatives I and II existed in subdomain IIA (site I) of HSA. Furthermore, derivatives were closed to tryptophan residues (Trp214). The results revealed that the intrinsic fluorescence of HSA could be successfully decreased by derivatives interactions. The experimental results were in agreement with molecular docking.