Combinatorial Evolution of Artificial Tyroserleutide Transporters for Enhanced Cancer Therapy In Vivo

<b>Peptide transporters are integral membrane proteins responsible for the cellular uptake of di- and tripeptides from the extracellular environment, which play pivotal roles in nutrient absorption, antigen presentation, and cellular signaling. Despite their essential biological functions, the development of artificial peptide transporters capable of efficiently transporting charge-neutral peptides remains elusive. Herein, we introduce a novel class of peptide transporters involving the combinatorial integration of anion and cation transport functionalities. Notably, 5F-C12, which functions as a molecular tweezer, forms a stable 1:1 complex with charge-neutral peptide tyroserleutide—an anticancer agent currently in Phase III clinical trials—and actively facilitates its transmembrane transport by shielding it from the membrane's hydrophobic core, with an EC</b>_{<strong>50</strong>}<b> value of 7.5 μM. Unprecedentedly, 5F-C12 could remarkably enhance the peptide’s bioavailability and exhibit a pronounced enhanced anticancer effect against MCF-7 breast cancer cells both </b><b><i>in vitro</i></b><b> and </b><b><i>in vivo</i></b><b>.</b>