Fig. 4. (a) Visualisation of the library through similarity matrix and heatmap generation with a side column indicating crosslinking to CAI (dark blue: hit) along with structures of non-crosslinking primary sulfonamides (9–10) and generalised structures of non-crosslinking secondary/tertiary sulfonamides (11a–r) and sulfones (12a–e). Dendrograms link structurally similar compounds and are a result of the hierarchical clustering method used. (b) Proposed iterative design–make–test hit ID protocol utilising the D2B HTC-PhABit platform to identify binders. (c) Crosslinking yields of the 100 HTC-PhABits with CAI (green: >1.5% crosslinking, blue: <1.5% crosslinking). (d) Heatmap displaying the similarity between the members of the 100-membered second-generation HTC-PhABit library with a side column indicating crosslinking to CAI (dark blue: hit). (e) Generalised structure of a hit primary sulfonamide summarising the tolerated features for binding in the Zn²⁺ binding pocket. (f) Structures and corresponding mass spectra for the 9 highest crosslinking hits from the single shot screen.