DNA-modularized construction of bivalent ligands precisely regulates receptor binding and activation

Great interest is focused on the construction of bivalent ligands, where the spacer is crucial to coordinate the distance and orientation of two pharmacophores for optimal biological effects. Current strategies rely on polymers as spacers but suffer from the paucity of structural precision and variability with time-consuming ligation procedures. Herein, we originate a DNA-modularized strategy where pharmacophores are modularly modified with the protecting groups used for automatic DNA synthesis while natural deoxynucleotides serve as spacers. By programmably regulating the number and permutation of the bridged DNA spacer, the two pharmacophores are adjusted with a defined distance and versatile orientation. Using this strategy, we successfully constructed a reservoir of bivalent ligands containing an orthosteric agonist and an allosteric modulator, exhibiting a single-nucleotide difference in the selective activation of muscarinic acetylcholine receptors. Our strategy opens a new avenue for the precise construction and efficient screening of bivalent ligands toward a myriad of biomedical applications.