Matteo Calvaresi

Chemophototherapy (CPT), combining chemotherapy and photodynamic therapy (PDT), is a promising strategy to enhance anticancer efficacy while reducing systemic toxicity. Many approved drugs contain chromophoric moieties that absorb visible light, suggesting their potential as intrinsic photosensitizers. Herein, we introduce a single-molecule chemophototherapy (SM-CPT) approach based on computational screening of approved anticancer drugs for repurposing as photosensitizers. A curated library of 238 small-molecule drugs was screened using time-dependent density functional theory to identify compounds with favourable photophysical properties. Fifteen candidates were identified, and mitoxantrone, pixantrone, daunorubicin, and epirubicin were selected for experimental validation. All compounds generated reactive oxygen species upon irradiation and showed enhanced cytotoxicity in breast cancer cell models, with up to an order-of-magnitude reduction in half-maximal inhibitory concentration values. The approach was further validated in ovarian cancer models, including platinum-resistant cells and patient-derived ascites, highlighting SM-CPT as a translationally attractive light-activated cancer therapy.

Conventional chemotherapeutics often suffer from poor aqueous solubility, short circulation half-lives, lack of tumor selectivity, and present dose-limiting systemic toxicity. This perspective examines the Trojan Horse Approach (THA), in which plasma carrier proteins (PCPs) are exploited as discrete, non-covalent supramolecular hosts for chemotherapeutic agents, and discusses its potential to overcome the limitations of both free drugs and conventional protein nanoparticles. Two principal strategies for assembling ex vivo drug–protein complexes are discussed: (i) molecular recognition, in which drugs occupy native ligand-binding pockets, and (ii) molecular anchoring, in which drugs are conjugated to high-affinity anchor ligands. The perspective critically assesses the current state of the art, distinguishes different levels of experimental validation, and discusses the major limitations and barriers to clinical translation. Structural, biochemical, and pharmacological evidence is examined for human serum albumin, transferrin, transthyretin, alpha-1-acid glycoprotein, alpha-fetoprotein, lactoferrin, and other plasma proteins. When appropriately designed and validated, protein-based complexes may preserve the native protein structure while conferring improved drug solubility, protection from premature metabolism, prolonged systemic circulation and enhanced tumor accumulation via combined passive and active targeting. PCPs represent a versatile and underexploited toolbox for cancer drug delivery. Expanding the repertoire of protein carriers, applying in silico rational design to guide the selection and optimization of drug–protein complexes, and exploring the use of patient-derived autologous proteins may open new opportunities for more effective and personalized therapies.

Photodynamic therapy (PDT) is a promising therapeutic modality based on the combined use of light and photosensitizers (PSs) and approved for treating several types of cancers. Thio‐heterocyclic naphthalimide‐based PSs are particularly attractive as phototheranostic agents, combining fluorescence imaging for diagnosis with reactive oxygen species (ROS)‐mediated cytotoxicity for treatment. However, no effective conjugation strategy for simultaneously endowing naphthalimides with water solubility and efficient ROS production ability currently exists. Herein, novel water‐soluble, heavy atom‐free thioxanthene‐fused naphthalimide–polyamine conjugates 1–4 have been developed. Structural variation in the polyamine moiety, including chain length and nitrogen content, allowed the fine‐tuning of key properties. Conjugates 2 and 3 exhibited (i) favorable photophysical and fluorescent properties, (ii) enhanced water solubility, (iii) improved photodynamic efficacy against MCF‐7 breast cancer cells, and (iv) negligible dark toxicity. This novel class of thioxanthene‐fused naphthalimide–polyamine conjugates provides a promising conjugation strategy for improving the drug‐like properties of PSs.

Water-soluble fullerene derivatives represent emerging molecular nanomaterials for antiviral drug discovery, yet their target selectivity against SARS-CoV-2 proteins remains insufficiently defined. Here, four functionalized [60]fullerenes, C60(OH)24, C60ser, C60ALA, and C60Trp, were evaluated as binders of key SARS-CoV-2 targets involved in viral entry and replication: spike glycoprotein, papain-like protease, 3CL main protease, and RNA-dependent RNA polymerase. Microscale thermophoresis revealed distinct target-dependent binding profiles governed by fullerene functionalization. The C60Trp fullerene was the most active derivative, binding Spike with nanomolar affinity (Kd = 94.8 nM) and showing micromolar affinity toward papain-like protease and 3CL main protease. Docking and molecular dynamics simulations supported the preferential interaction of C60Trp with the spike protein cavity through combined fullerene cage complementarity and polar interactions with lysine and arginine residues. The tested derivatives showed low cytotoxicity in A549 cells, and C60Trp was efficiently internalized. These findings identify tryptophan-functionalized [60]fullerene as a promising multivalent scaffold for SARS-CoV-2 protein targeting.

Here we present three novel photoswitchable DNA intercalators based on the diazocine scaffold. These compounds bind B-DNA in the trans state (K_D,trans = (4.14 ± 1.25)-(7.84 ± 3.41) × 10^-4 M) and show fully reversible "on"-"off" behavior upon visible-light irradiation without any sign of fatigue over six cycles. The affinity of the trans state is at least 1000-fold higher than of the cis state. The ability to reversibly turn off all observable binding with in situ visible light irradiation is so far unmatched in the literature. Preliminary cytotoxicity studies of 3,3'-dimethanamine-diazocine (3) show negligible dark toxicity (EC₅₀ > 100 μM) with a significant increase upon irradiation (395 nm, EC₅₀ = 45.4 μM, PI > 2) in HeLa cells, allowing for spatiotemporal control of activity in photopharmacology applications.

The blood‐brain barrier (BBB) is a highly selective gatekeeper. While providing essential protection against pathogens, BBB also poses a major challenge by restricting the delivery of systemically administered therapeutics. Consequently, there is a pressing need for innovative strategies that can efficiently and selectively target neuronal populations to treat diverse brain disorders. Nanomaterials enable transcellular transport across the endothelial layer; however, given the biomolecular corona paradigm, designing biomimetic hybrid nanoconstructs that preserve and leverage biological functionality within a complex physiological environment is more promising. Here, we investigated the potential of M13 bacteriophages as biocompatible and versatile nanovectors across the BBB. M13 phages exhibited: ( i ) a high cargo capacity within the main protein capsomer (here exploited for fluorescent labeling); ( ii ) offered genetic flexibility for ligand display; ( iii ) showed a natural propensity to efficiently cross the endothelial layer via specific intracellular pathways; ( iv ) maintained their structure and functionality intact. We also demonstrate that M13 phages engineered to target specific neuronal populations via single‐domain antibody display retain their specificity after BBB translocation and in a protein‐ and cell‐crowded environment. Overall, our results provide mechanistic insights emphasizing an unprecedented field of application for (engineered) M13 phages as biomimetic nanotools for targeting and delivery across the BBB.

Bioconjugation of Rose Bengal and Chlorin e6 onto an FRα-targeted M13 bacteriophage enables validated dual-light phototheranostic assessment and selective treatment of ovarian cancer.

Perfluoroalkyl substances (PFAS) are a family of over seven million chemicals found in a vast number of industrial and consumer applications. Often referred to as the “forever chemicals,” they have gained increasing attention due to their environmental and health implications. The growing awareness and interest in these perfluorinated pollutants demand the exploitation of an integrated approach where the use of computational pipelines can play a major role in the understanding and prediction of their behavior. To this end, we developed an accurate and transferable coarse-grain PFAS model in the framework of the Martini 3 force field. Given the large amount of PFAS already reported, 18 linear perfluorocarboxylic acids and perfluorosulfonic acids of different chain lengths were considered in the parametrization. The model was validated following the standard Martini procedure and complemented with additional studies on self-aggregation and the interaction with graphene, which is a common substrate for sensors and wastewater remediation adsorbents. The results are highly consistent with both all-atom simulations and experimental data, successfully reproducing the key structural and physical properties. We believe that our study opens the way for high-throughput simulations to explore the interaction between PFAS and nanoparticles/(bio)molecules.

X-MACE predicts key excited-state properties with near quantum-mechanical accuracy at low computational cost, enabling rapid screening of molecular libraries. Its best candidate was validated in vitro for ROS generation and enhanced phototoxicity.