This publication makes a speciality of thielocin B1 (TB1), which was once came upon to be an inhibitor of protein–protein interactions (PPIs) of proteasome assembling chaperone (PAC) three homodimer, and elucidates the mechanism through nuclear magnetic resonance (NMR) experiences. Interfaces of PPIs lately were anticipated to be novel healing objectives, whereas it's tough to use traditional technique in response to lock and key thought. the writer completed the 1st overall synthesis of TB1 and its spin-labeled spinoff to hold out NMR experiments as the provide of TB1 from typical resources was once constrained. exact 2,2’,6,6’-tetrasubstituted diphenyl ether moiety of TB1 used to be synthesized from a depsidone skeleton through chemoselective relief of lactone. within the means of elongating part wings, effective formylation using dichloromethyl methyl ether–silver trifluoromethanesulfonate was once constructed for the sterically hindered fragrant compound. NMR titration experiments and paramagnetic leisure enhancement remark of PAC3 homodimer have been played with synthesized TB1 and its molecular probe, respectively. the result of the above NMR reviews and extra in silico docking reports instructed that TB1 promotes the dissociation to monomeric PAC3 after interplay with PAC3 homodimer. The infrequent mechanism proven during this e-book exhibits a possible novel drug goal within the interfaces of PPIs without hollow space or groove.