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The predicted inhibitor-binding mode of 1j with an orientation and conformation that favors most negative docking score when superimposed with the endogenous substrate NCA of the NNMT enzyme indicated that the analog binds consistent with the binding mode of NCA, i.e., the N1-atom of both ligands aligned almost identical, conferring similar molecular interactions with key residues within the active site of the enzyme

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Additional biochemical assays were used to test the ability of compounds to inhibit nicotinamide phosphoribosyl transferase (NAMPT) and NAD+-dependent protein deacetylase sirtuin 1 (SIRT1), two enzymes in the NAD+ biosynthesis/salvage pathway