Effective and Selective Anti‐Cancer Protein Delivery via All‐Functions‐in‐One Nanocarriers Coupled with Visible Light‐Responsive, Reversible Protein Engineering

Abstract

Efficient intracellular delivery of protein drugs and tumor-specific activation of protein functions are critical toward anti-cancer protein therapy. However, an omnipotent protein delivery system that can harmonize the complicated systemic barriers as well as spatiotemporally manipulate protein function is lacking. Herein, an "all-functions-in-one" nanocarrier doped with photosensitizer (PS) is developed and coupled with reactive oxygen species (ROS)-responsive, reversible protein engineering to realize cancer-targeted protein delivery, and spatiotemporal manipulation of protein activities using long-wavelength visible light (635 nm) at low power density (5 mW cm −2 ). Particularly, RNase A is caged with H 2 O 2 -cleavable phenylboronic acid to form 4-nitrophenyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl carbonate (NBC)-modified RNase (RNBC), which is encapsulated in aciddegradable, ketal-crosslinked PEI (KPEI)-based nanocomplexes (NCs) coatedwith PS-modified hyaluronic acid (HA). Such NCs harmonize the critical processes for protein delivery, wherein HA coating renders NCs with long blood circulation and cancer cell targeting, and KPEI enables endosomal escape as well as acid-triggered intracellular RNBC release. Tumor-specific light irradiation generates H 2 O 2 to kill cancer cells and restore the protein activity, thus achieving synergistic anti-cancer efficacy. It is the first time to photomanipulate protein functions by coupling ROS-cleavable protein caging with PS-mediated ROS generation, and the "all-functions-in-one" nanocarrier represents a promising example for the programmed anti-cancer protein delivery.

Nanosized Delivery Systems for Therapeutic Proteins: Clinically Validated Technologies and Advanced Development Strategies

“…Nanosized dendrimers and hyperbranched polymers have also been proposed as protein nanocarriers. Negatively charged proteins can be easily entrapped within positively charged dendrimers such as PAMAM (He et al, 2018). Dendrimer-based carriers with a hydrophobic membranedisruptive region (aromatic motif), and a multivalent protein binding surface (guanidyl-based) was developed for the delivery of BSA, R-phycoerythrin, p53, saporin, β-galactosidase, and peptides into the cytosol of living cells (Chang et al, 2017;Liu et al, 2019).…”

Size Switchable Nanoclusters Fueled by Extracellular ATP for Promoting Deep Penetration and MRI‐Guided Tumor Photothermal Therapy

“…In the past few decades, proteins or protein‐based theranostic agents (PBTAs) have been extensively explored for the diagnosis and treatment of various cancers . Therapeutic proteins, such as GOx, DNase I, RNase A, cytochrome c, or β‐Gal, as bio‐macromolecular drugs, exhibited exciting activity on antitumor therapy. Other biocompatible proteins, like albumin, hemoglobin, or transferrin, could acted as the carrier of therapeutic small molecular drugs, dyes or the diagnosis agent.…”

Dynamic Covalent Bond‐Assisted Programmed and Traceless Protein Release: High Loading Nanogel for Systemic and Cytosolic Delivery

“…In this regard, many systems are not suitable for in vivo uses, especially when systemically administrated. Moreover, because of the contradictory requirements posed by the complex biological barriers to the design of nanomedicine, very few examples, regardless the covalent or noncovalent approach, can realize efficient systemic and cytosolic protein delivery . As a result, only those proteins do not require an intracellular mechanism of action are currently used in clinic.…”

“…To this end, one unmet need is a simple formulation simultaneously enabling high protein loading, outstanding stability under physiological conditions, and most importantly, a traceless release character in the cytoplasm. [12b,14–16]…”

“…Third, we have recently shown that the onset pH of the maleamic amide hydrolysis can be precisely adjusted to both the extracellular tumor microenvironment (pH e , ≈6.5–6.8) and the intracellular late endosome (pH i , ≈5.0–6.0) by using bis‐ and monosubstituted MA derivatives, respectively ( Figure ). [17a,18] Thus, the introduction of both bis‐and monosubstituted MA in the NG allows programmed cargo release responding to tandem pH changes . Previously, MA derivatives have been widely used as charge inversion moieties in nanomedicines facilitating enhanced tumor retention and cellular uptake at tumor microenvironment .…”