The fluorination effect of fluoroamphiphiles in cytosolic protein delivery

Abstract

Direct delivery of proteins into cells avoids many drawbacks of gene delivery, and thus has emerging applications in biotherapy. However, it remains a challenging task owing to limited charges and relatively large size of proteins. Here, we report an efficient protein delivery system via the co-assembly of fluoroamphiphiles and proteins into nanoparticles. Fluorous substituents on the amphiphiles play essential roles in the formation of uniform nanoparticles, avoiding protein denaturation, efficient endocytosis, and maintaining low cytotoxicity. Structure-activity relationship studies reveal that longer fluorous chain length and higher fluorination degree contribute to more efficient protein delivery, but excess fluorophilicity on the polymer leads to the pre-assembly of fluoroamphiphiles into stable vesicles, and thus failed protein encapsulation and cytosolic delivery. This study highlights the advantage of fluoroamphiphiles over other existing strategies for intracellular protein delivery.

Fluorinated Polymer Mediated Transmucosal Peptide Delivery for Intravesical Instillation Therapy of Bladder Cancer

“…The preparation of MPI/F‐PEI NPs was started with the synthesis of F‐PEI, which was conducted following the literature protocol by using branched PEI with a molecular weight of 25 kDa as the polymer scaffold and 3‐(perfluorohex‐1‐yl)‐1,2‐propenoxide as the fluorous tag ( Figure a) . According to the fluorine element analysis, the average number of fluoroalkane conjugated on each PEI was ≈30 in the obtained F‐PEI.…”

“…In recent years, it has been discovered that fluorinated polymers could be utilized for rather effective transmembrane transportation of biomacromolecules such as genes and proteins, by utilizing the unique hydrophobic and lipohphobic behaviors of fluorocarbon chains to endow efficient cross‐membrane penetration, making them particularly promising for applications in gene therapy of cancer . In our previous studies, we synthesized a series of fluorinated polymers and explored its structure–activity relationship on biomolecular transportation.…”

“…The results demonstrated that higher fluorination degree contributed to higher biomolecule delivery efficiency, while excess fluorophilicity on the amphiphiles resulted in the preassembly of fluoroamphiphiles, and thus impeded biomolecular encapsulation and intracellular delivery. In that work, fluorinated polyethylenimine (F‐PEI) with proteins showed remarkable advantages in self‐assembly property, endocytosis, endosomal escape, biocompatibility, and intracellular release . Therefore, exactly the same formulation of F‐PEI was chosen in our current study.…”

“…In this work, we ought to apply F‐PEI as a new type of transmucosal carrier for the delivery of MPI to treat orthotopic bladder cancer by intravesical instillation therapy ( Scheme a). Owing to the fluorophilic effect, hydrogen‐bonding, and electrostatic interactions, MPI peptide upon mixing with F‐PEI or nonfluorinated PEI would form MPI/F‐PEI or MPI/PEI nanoparticles (NPs) by self‐assembly. Compared to free MPI and MPI/PEI NPs, MPI/F‐PEI showed dramatically enhanced cross‐membrane, transmucosal and intratumoral penetration abilities as evidenced by a number of different in vitro and ex vivo assays (Scheme b).…”

Mixed Fluorinated/Hydrogenated Self‐Assembled Monolayer‐Protected Gold Nanoparticles: In Silico and In Vitro Behavior

“…Spontaneous phase separation in supramolecular aggregates of F/H amphiphiles is well documented for several such systems (e.g., micelles, Langmuir, and Langmuir–Blodgett films), including 2D SAMs on gold surfaces . Furthermore, fluorinated amphiphiles are receiving increasing interest for biological/biomedical applications as they confer useful properties to the materials they are linked to, such as inertness, facilitated cellular uptake, avoidance of protein denaturation, and reduced toxicity . Additionally, being NMR active, fluorine nuclei also assist in probing biological interactions and may lead to novel supramolecular contrast agents for bioimaging .…”