Palladium responsive liposomes for triggered release of aqueous contents.

Palladium (Pd) is a promising metal catalyst for novel bioorthogonal chemistry and prodrug activation. This report describes the first example of palladium responsive liposomes. The key molecule is a new caged phospholipid called Alloc-PE that forms stable liposomes (large unilamellar vesicles, ∼220 nm diameter). Liposome treatment with PdCl2 removes the chemical cage, liberates membrane destabilizing dioleoylphosphoethanolamine (DOPE), and triggers liposome leakage of encapsulated aqueous contents. The results indicate a path towards liposomal drug delivery technologies that exploit transition metal triggered leakage.

Intracellular fluorescence competition assay for inhibitor engagement of histone deacetylase.

An intracellular fluorescence competition assay was developed to assess the capability of inhibitor candidates to engage histone deacetylase (HDAC) inside living cells and thus diminish cell uptake and staining by the HDAC-targeted fluorescent probe APS. Fluorescence cell microscopy and flow cytometry showed that pre-incubation of living cells with candidate inhibitors led to diminished cell uptake of the fluorescent probe. The assay was effective because the fluorescent probe (APS) possessed the required performance properties, including bright fluorescence, ready membrane diffusion, selective intracellular HDAC affinity, and negligible acute cytotoxicity. The concept of an intracellular fluorescence competition assay is generalizable and has broad applicability since it obviates the requirement to use the isolated biomacromolecule target for screening of molecular candidates with target affinity.

Supramolecular Loading of a Broad Spectrum of Molecular Guests In Hyperbranched Polytriazole Nanoparticles with Cores Containing Multiple Functional Groups.

This study evaluated the supramolecular properties of a new family of water-soluble hyperbranched polytriazoles that have a unimolecular micelle structure. Two new, structurally related hyperbranched polymers (HBPa and HBPn), with the same size (Dh = 11 nm) and core-shell architecture, were prepared and found to act as nanoscale hosts for a broad spectrum of molecular guests. The globular-shaped hyperbranched polymers were synthesized by a straightforward one-pot polymerization method that permits easy synthetic control of the multiple functional groups within the core. Surrounding the core is a shell of polyethylene glycol chains that promotes solubility in pH 7.4 buffer solution and inhibits self-aggregation of the nanoparticles. The core of HBPa, containing a mixture of anionic carboxylate groups and 1,2,3-triazole rings, could be loaded with cationic hydrophilic (i.e., propidium iodide) or partially hydrophobic (i.e., Hoechst 33342) dyes or drugs, including a binary dye/drug pair (i.e., indocyanine green/mitoxantrone). The core of HBPn, containing a mixture of uncharged 2-pentanone chains and 1,2,3-triazole rings, could be loaded with uncharged and very hydrophobic dyes (i.e., Nile Red) or drugs. Improved aqueous solubility of camptothecin was achieved 10-fold from 8.4 to 75 ng/mL. Additionally, cell toxicity studies showed that HBPn was able to release the camptothecin drug inside A549 cancer cells resulting in increased cell death. Taken together, the results suggest that this new family of water-soluble hyperbranched polytriazoles could be broadly useful as nanocarriers for various applications in therapy, imaging, or a combination of the two (theranostics).

Fluorescent Self-Threaded Peptide Probes for Biological Imaging.

A general synthetic method creates a new class of covalently connected, self-threaded, fluorescent molecular probes with figure-eight topology, an encapsulated deep-red fluorophore, and two peripheral peptide loops. The globular molecular shape and rigidified peptide loops enhance imaging performance by promoting water solubility, eliminating probe self-aggregation, and increasing probe stability. Moreover, the peptide loops determine the affinity and selectivity for targets within complex biological samples such as cell culture, tissue histology slices, or living subjects. For example, a probe with cell-penetrating peptide loops targets the surface of cell plasma membranes, whereas, a probe with bone-targeting peptide loops selectively stains the skeleton within a living mouse. The unique combination of bright deep-red fluorescence, high stability, and predictable peptide-based targeting is ideal for photon intense fluorescence microscopy and biological imaging.

Supramolecular capture of highly polar amidosquaraine dye in water with nanomolar affinity and large turn-on fluorescence.

A supramolecular dye-capture system comprising anionic amidosquaraine guest and macrocyclic tetralactam host exhibits nanomolar affinity and "turn on" visible fluorescence. Utility is demonstrated with a new fluorescent assay for liposome leakage induced by the biomedically important enzyme phospholipase A2.