Bacterial fouling on surfaces significantly increases the resistance of bacteria toward antibiotics, which leads to medical complications and a corresponding financial burden. Here, we report on a general and robust technique for facile modification of various surfaces with different antibacterial agents. Our approach in this study was inspired by the strong adhesion of mussel adhesion proteins (MAPs) to many types of surfaces, including metals, polymers, and inorganic materials. Thus, glass and polymeric slides were dip-coated with dopamine, as a MAP mimic, and the resulting surfaces were characterized. The reactivity of dopamine-coated surfaces toward nucleophilic addition was then confirmed by reacting them with fluorescent probes containing either a free amino or a free thiol group. Laser scanning confocal microscopy (LSCM), X-ray photoelectron spectroscopy (XPS), confocal Raman microscopy, matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectroscopy, and cyclic voltammetry studies collectively suggested that the probes had covalently attached to the surfaces. Fabrication of dopamine-coated surfaces with an antibacterial quaternary amine or an ultrashort lipopeptide analog generated surfaces that effectively kill Escherichia coli and Staphylococcus aureus cells on contact. Moreover, minimal leaching of the fabricated agent was detected after prolonged incubation. This technique could be further developed to a ''paint-like'' or selfassembling monolayer-like procedure for the preparation of antibacterial surfaces on various materials.

Single-triggered disassemble dendrimers were recently developed and introduced as a potential platform for a multi-prodrug. These unique structural dendrimers can release all of their tail units through a self-immolative chain fragmentation initiated by a single cleavage at the dendrimer's core. There are several examples for the bioactivation of first-generation self-immolative dendritic prodrugs. However, enzymatic activation failed for second-generation self-immolative dendrimers. The hydrophobic large molecular structure of the dendritic prodrugs results in aggregation under aqueous conditions and prevented the enzyme from reaching the triggering substrate. Here we show a simple solution for the enzymatic activation of second-generation self-immolative dendrimers. Poly(ethylene glycol) (PEG) was conjugated to the dendritic platform via click chemistry. The poly(ethylene glycol) tails significantly decreased the hydrophobic properties of the dendrimers and thereby prevented aggregate formation. We designed and synthesized a dendritic prodrug with four molecules of the anticancer agent camptothecin and a trigger that can be activated by penicillin-G-amidase. The PEG5000-conjugated, self-immolative dendritic prodrug was effectively activated by penicillin-G-amidase under physiological conditions and free camptothecin was released to the reaction media. Cell-growth inhibition assays demonstrated increased toxicity of the dendritic prodrug upon incubation with the enzyme.

The M-type K ϩ current (M-current), encoded by Kv7.2/3 (KCNQ2/3) K ϩ channels, plays a critical role in regulating neuronal excitability because it counteracts subthreshold depolarizations. Here we have characterized the functions of pre-and postsynaptic M-channels using a novel Kv7.2/3 channel opener, NH6, which we synthesized as a new derivative of N-phenylanthranilic acid. NH6 exhibits a good selectivity as it does not affect Kv7.1 and I KS K ϩ currents as well as NR1/NR2B, AMPA, and GABA A receptor-mediated currents. Superfusion of NH6 increased recombinant Kv7.2/3 current amplitude (EC 50 ϭ 18 M) by causing a hyperpolarizing shift of the voltage activation curve and by markedly slowing the deactivation kinetics. Activation of native M-currents by NH6 robustly reduced the number of evoked and spontaneous action potentials in cultured cortical, hippocampal and dorsal root ganglion neurons. In hippocampal slices, NH6 decreased somatically evoked spike afterdepolarization of CA1 pyramidal neurons and induced regular firing in bursting neurons. Activation of M-channels by NH6, potently reduced the frequency of spontaneous excitatory and inhibitory postsynaptic currents. Activation of M-channels also decreased the frequency of miniature excitatory (mEPSC) and inhibitory (mIPSC) postsynaptic currents without affecting their amplitude and waveform, thus suggesting that Mchannels presynaptically inhibit glutamate and GABA release. Our results suggest a role of presynaptic M-channels in the release of glutamate and GABA. They also indicate that M-channels act pre-and postsynaptically to dampen neuronal excitability.

"Chemical adaptor systems" are molecules used to link different functionalities, based on unique reactivity that allows controlled fragmentation. Two different mechanistic reactivities were used to prepare chemical adaptor systems. The first is based on a spontaneous intra-cyclization reaction to form a stable ring molecule. Cleavage of the trigger generates a free nucleophile, for example, an amine group, which undergoes intra-cyclization to release the target molecule from the handle part (e.g., a targeting antibody or a solid support for synthesis). The second applied reactivity is an elimination reaction, which is usually based on a quinone-methide-type rearrangement. Similarly, cleavage of the trigger generates a free phenol functionality, which can undergo a self-elimination reaction through a quinone-methide rearrangement to release the target molecule. The adaptor molecules have been applied in the field of drug delivery to release a drug from a targeting device and in the field of solid-phase synthesis to release a synthetic molecule from the solid support. A chemical adaptor molecule has also been used as a building unit to construct dendrimers with a triggered fragmentation.

Cyclooxygenase (COX) enzymes are molecular targets of nonsteroidal anti-inflammatory drugs (NSAIDs), the most used medication worldwide. However, the COX enzymes are not the sole molecular targets of NSAIDs. Recently, we showed that two NSAIDs, diclofenac and meclofenamate, also act as openers of Kv7.2/3 K+ channels underlying the neuronal M-current. Here we designed new derivatives of diphenylamine carboxylate to dissociate the M-channel opener property from COX inhibition. The carboxylate moiety was derivatized into amides or esters and linked to various alkyl and ether chains. Powerful M-channel openers were generated, provided that the diphenylamine moiety and a terminal hydroxyl group are preserved. In transfected CHO cells, they activated recombinant Kv7.2/3 K+ channels, causing a hyperpolarizing shift of current activation as measured by whole-cell patch-clamp recording. In sensory dorsal root ganglion and hippocampal neurons, the openers hyperpolarized the membrane potential and robustly depressed evoked spike discharges. They also decreased hippocampal glutamate and GABA release by reducing the frequency of spontaneous excitatory and inhibitory post-synaptic currents. In vivo, the openers exhibited anti-convulsant activity, as measured in mice by the maximal electroshock seizure model. Conversion of the carboxylate function into amide abolished COX inhibition but preserved M-channel modulation. Remarkably, the very same template let us generating potent M-channel blockers. Our results reveal a new and crucial determinant of NSAID-mediated COX inhibition. They also provide a structural framework for designing novel M-channel modulators, including openers and blockers.