Exploring the Effect of Amphiphile Architecture on Intracellular Protein Delivery Capacity: Dimeric versus Trimeric Amphiphiles

Carrier-mediatedintracellular protein delivery holdstremendousapplication potential in biology and medicine. The ideal carrier shouldbe well-controlled and cost-effective and able to facilitate robustdelivery of diverse types of proteins into the target cells, thusensuring efficacy in different application scenarios. Here, we describea modular chemistry approach for generating a small-molecule amphiphilemolecular library based on the Ugi four-component reaction under one-potand mild conditions. Then, two different types of amphiphiles withthe dimeric or trimeric architecture were obtained for intracellularprotein delivery through in vitro screening test.Depending on the precise adjustment of the hydrophobic tails of amphiphiles,the optimized trimeric amphiphile (TA) exhibited more superior proteinloading performance and a higher efficiency of delivering proteinsinto cells through the endocytosis pathway and subsequent endosomalescape. Furthermore, we demonstrated that the TA could be a universaldelivery carrier capable of transporting broad-spectrum proteins,especially for the hard-to-deliver native antibodies, into the cytosol.Overall, we describe a robust amphiphile platform with a well-definedand cost-effective design to improve the cytosolic protein deliverycapacity, exhibiting great promise for developing intracellular protein-basedtherapeutics.

Automated summary

A small-molecule amphiphile molecular library was generated and screened, resulting in the identification of dimeric and trimeric amphiphiles for intracellular protein delivery. The trimeric amphiphile showed superior protein loading performance and delivery efficiency through the endocytosis pathway and subsequent endosomal escape. Moreover, the trimeric amphiphile was capable of delivering hard-to-deliver native antibodies into the cytosol. Overall, this well-designed and cost-effective amphiphile platform holds great promise for improving cytosolic protein delivery capacity.