Fig. 3: Screening of macrocyclic compound library against thrombin, hit identification, and structural analysis. | Nature Communications

Fig. 3: Screening of macrocyclic compound library against thrombin, hit identification, and structural analysis.

From: Synthesis and direct assay of large macrocycle diversities by combinatorial late-stage modification at picomole scale

Fig. 3

a Heat map with thrombin inhibition indicated for each macrocycle. The amino acid sequences of the scaffolds are provided in the Source Data file. b Replicate reaction (black) compared to original screen (green) of all macrocycles containing acid 14. For each of the two independent experiments, the reactions were performed once and the thrombin inhibition was measured once. c Chemical structures and activities of the top three hits M1, M2, and M3. The mean values and SDs of three independent measurements are shown. d Chromatographic separation of the acylation reactions to generate M1 and M2 and analysis of the fractions for thrombin inhibition. e X-ray structure of M1 bound to human thrombin at 2.27 Å resolution (PDB 6Z48) (https://www.rcsb.org/structure/6Z48). The inhibitor is shown in the space-filling model with the scaffold in blue and the carboxylic acid in yellow. f Zoomed in structure of thrombin with the sub-sites indicated. The chlorothiophene group fills the S1 sub-site and forms an interaction with Tyr228.

Back to article page