A triple-stimulus responsive melanin-based nanoplatform with an aggregation-induced emission-active photosensitiser for imaging-guided targeted synergistic phototherapy/hypoxia-activated chemotherapy.

Multimodal synergistic therapy has gained increasing attention in cancer treatment to overcome the limitations of monotherapy and achieve high anticancer efficacy. In this study, a synergistic phototherapy and hypoxia-activated chemotherapy nanoplatform based on natural melanin nanoparticles (MPs) loaded with the bioreduction prodrug tirapazamine (TPZ) and decorated with hyaluronic acid (HA) was developed. A self-reporting aggregation-induced emission (AIE)-active photosensitizer (PS) (BATTMN) was linked to the prepared nanoparticles by boronate ester bonds. The MPs and BATTMN-HA played roles as quenchers for PS and cancer targeting/photodynamic moieties, respectively. As a pH sensitive bond, the borate ester bonds between HA and BATTMN are hydrolysed in the acidic cancer environment, thereby separating BATTMN from the nanoparticles and leading to the induction of fluorescence for imaging-guided synergistic phototherapy/hypoxia-activated chemotherapy under dual irradiation. TPZ can be released upon activation by pH, near-infrared (NIR) and hyaluronidase (Hyal). Particularly, the hypoxia-dependent cytotoxicity of TPZ was amplified by oxygen consumption in the tumor intracellular environment induced by the AIE-active PS in photodynamic therapy (PDT). The nanoparticles developed in our research showed favorable photothermal conversion efficiency (η = 37%), desired cytocompatibility, and excellent synergistic therapeutic efficacy. The proposed nanoplatform not only extends the application scope of melanin materials with AIE-active PSs, but also offers useful insights into developing multistimulus as well as multimodal synergistic tumor treatment.

Protein Substrates for Reaction Discovery: Site-Selective Modification with Boronic Acid Reagents.

Chemical modification of natural proteins must navigate difficult selectivity questions in a complex polyfunctional aqueous environment, within a narrow window of acceptable conditions. Limits on solvent mixtures, pH, and temperature create challenges for most synthetic methods. While a protein's complex polyfunctional environment undoubtedly creates challenges for traditional reactions, we wondered if it also might create opportunities for pursuing new bioconjugation reactivity directly on protein substrates. This Account describes our efforts to date to discover and develop new and useful reactivity for protein modification by starting from an open-ended screen of potential transition-metal catalysts for boronic acid reactivity with a model protein substrate. By starting from a broad screen, we were hoping to take advantage of the very many potential reactive sites on even a small model protein. And perhaps more importantly, whole proteins as reaction screening substrates might exhibit uniquely reactive local environments, the results of a dense combination of functional groups that would be nearly impossible to mimic in a small-molecule context. This effort has resulted in the discovery of four new protein modification reactions with boronic acid reagents, including a remarkable modification of specific backbone N-H bonds. This histidine-directed Chan-Lam coupling, based on specific proximity of an imidazole and two amide groups, is one important example of powerful reactivity that depends on a combination of functional groups that proteins make possible. Other bioconjugation reactions uncovered include a three-component tyrosine metalation with rhodium(III), a nickel-catalyzed cysteine arylation, and an unusual ascorbate-mediated oxidative process for N-terminal modification. The remarkably broad scope of reactivity types encountered in this work is a testament to the breadth of boronic acid reactivity. It is also a demonstration of the diverse reactivities that are possible by the combined alteration of boronic acid structure and metal promoter. The discovery of specific backbone modification chemistry has been a broadly empowering reactivity. Pyroglutamate, a naturally occurring posttranslational modification, exhibits remarkably high reactivity in histidine-directed backbone modification, which allows us to treat pyroglutamate as a reactive bioorthogonal handle that is readily incorporated into proteins of interest by natural machinery. In another research direction, the development of a vinylogous photocleavage system has allowed us to view backbone modification as a photocaging modification which is released by exposure to light.

A simple boronic acid-based fluorescent probe for selective detection of hydrogen peroxide in solutions and living cells.

An approach of high sensitivity and selectivity for hydrogen peroxide (H2O2) detection is highly demanded due to its important roles in regulating diverse biological process. In this work, we introduced an easily synthesized fluorescent "turn off" probe, BNBD. It is designed based on the core structure of 4-chloro-7-nitrobenzofurazan as a fluorophore and incorporated with a specific H2O2-reactive group, aryl boronate, for sensitive and selective detection of H2O2. We demonstrated its selectivity by incubating the probe with other types of ROS, and measured the limit of detection of BNBD as 1.8 nM. BNBD is also conducive to H2O2 detection at physiological conditions. We thus applied it to detect both exogenous and endogenous changes of H2O2 in living cells by confocal microscopy, supporting its future applications to selectively monitor H2O2 levels and identify H2O2-related physiological or pathological responses from live cells or tissues in the near future.

Acceleration of arylzinc formation and its enantioselective addition to aldehydes by microwave irradiation and aziridine-2-methanol catalysts.

The formation and 2-amino alcohol catalyzed addition of arylzinc reagents from and with boronic acids, respectively, is drastically accelerated to a few minutes under microwave irradiation without loss of enantioselectivity (up to 98% ee). Of the amino acid derived catalysts tested, the conformationally restricted bulky substituted aziridine-2-methanols derived from serine show the best overall performance in the formation of diarylmethanols.

The Suzuki homocoupling reaction under high-intensity ultrasound.

The Pd-catalysed Suzuki homocoupling of boronic acids was successfully carried out in water under high-intensity ultrasound (HIU). Heterogeneous catalysis with Pd/C, facilitating work up and purification, could be used without adding phosphine ligands. Reaction rates and yields were strongly influenced by the oxidant employed; excellent results were obtained using either molecular oxygen or 3-bromo-4-hydroxycoumarin. 3-Arylation of the latter with the Suzuki procedure had failed, exclusively affording the homocoupling products, symmetric biaryls. Besides offering a number of operational advantages, the use of HIU broadens the field of application for the Suzuki reaction.

Ultrasound effect on Suzuki reactions. 1. Synthesis of unsymmetrical biaryls.

Ultrasound effect on the Pd(0) catalysed reaction of arylboronic acid with halobenzenes was investigated. The effect of catalyst, base as well as solvent was tested. Heterogenous reaction of iodoarenes with different arylboronic acids, catalysed by Pd/C and KF as the base in methanol:water mixture resulted in good yields of cross-coupling products. Reaction time of sonochemical reaction was 1 h, while 4 h of reflux was necessary to achieve comparable results. Bromobenzenes gave best results using aqueous solution of PdCl2 as the catalyst, potassium carbonate as the base in toluene:water two phase system using TEBA (benzyltriethylammonium chloride) as PT catalyst. Chlorobenzenes gave just feeble yields of cross-coupling products.