Comparison and Mechanism Study of Antibacterial Activity of Cationic and Neutral Oligo-Thiophene-Ethynylene.

Photodynamic therapy (PDT) is a potential approach to resolve antibiotic resistance, and phenylene/thiophene-ethynylene oligomers have been widely studied as effective antibacterial reagents. Oligomers with thiophene moieties usually exhibit good antibacterial activity under light irradiation and dark conditions. In the previous study, we verified that neutral oligo-p-phenylene-ethynylenes (OPEs) exhibit better antibacterial activity than the corresponding cationic ones; however, whether this regular pattern also operates in other kinds of oligomers such as oligo-thiophene-ethynylene (OTE) is unknown. Also, the antibacterial activity comparison of OTEs bearing cyclic and acyclic amino groups will offer useful information to further understand the role of amino groups in the antibacterial process and guide the antibacterial reagent design as amino groups affect the antibacterial activity a lot. We synthesized four OTEs bearing neutral or cationic, cyclic, or acyclic amino groups and studied their antibacterial activity in detail. The experimental results indicated that the OTEs exhibited better antibacterial activity than the OPEs, the neutral OTEs exhibited better antibacterial activity in most cases, and OTEs bearing cyclic amino groups exhibited better antibacterial activity than those bearing acyclic ones in most cases. This study provides useful guidelines for further antibacterial reagent design and investigations.

Chemical Diversification of Simple Synthetic Antibodies.

Antibodies possess properties that make them valuable as therapeutics, diagnostics, and basic research tools. However, antibody chemical reactivity and covalent antigen binding are constrained, or even prevented, by the narrow range of chemistries encoded in canonical amino acids. In this work, we investigate strategies for leveraging an expanded range of chemical functionality using yeast displayed antibodies containing noncanonical amino acids (ncAAs) in or near antibody complementarity determining regions (CDRs). To enable systematic characterization of the effects of ncAA incorporation on antibody function, we first investigated whether diversification of a single antibody loop would support the isolation of binding clones against immunoglobulins from three species. We constructed and screened a billion-member library containing canonical amino acid diversity and loop length diversity only within the third complementarity determining region of the heavy chain (CDR-H3). Isolated clones exhibited moderate affinities (double- to triple-digit nanomolar affinities) and, in several cases, single-species specificity, confirming that antibody specificity can be mediated by a single CDR. This constrained diversity enabled the utilization of additional CDRs for the installation of chemically reactive and photo-cross-linkable ncAAs. Binding studies of ncAA-substituted antibodies revealed that ncAA incorporation is reasonably well tolerated, with observed changes in affinity occurring as a function of ncAA side chain identity, substitution site, and the ncAA incorporation machinery used. Multiple azide-containing ncAAs supported copper-catalyzed azide-alkyne cycloaddition (CuAAC) and strain-promoted azide-alkyne cycloaddition (SPAAC) without the abrogation of binding function. Similarly, several alkyne substitutions facilitated CuAAC without the apparent disruption of binding. Finally, antibodies substituted with a photo-cross-linkable ncAA were evaluated for ultraviolet-mediated cross-linking on the yeast surface. Competition-based assays revealed position-dependent covalent linkages, strongly suggesting successful cross-linking. Key findings regarding CuAAC reactions and photo-cross-linking on the yeast surface were confirmed using soluble forms of ncAA-substituted clones. The consistency of findings on the yeast surface and in solution suggest that chemical diversification can be incorporated into yeast display screening approaches. Taken together, our results highlight the power of integrating the use of yeast display and ncAAs in search of proteins with "chemically augmented" binding functions. This includes strategies for systematically introducing small molecule functionality within binding protein structures and evaluating protein-based covalent target binding. The efficient preparation and chemical diversification of antibodies on the yeast surface open up new possibilities for discovering "drug-like" protein leads in high throughput.

Stable and Well-Organized Near-Infrared Platinum(II)-Acetylide-Based Metallacycles-Mediated Cancer Phototherapy.

The development of metallacycles with high stability and intense near-infrared (NIR) absorption is important for biomedical applications. However, very few molecular design strategies have been developed on such metallacycles. Herein, we report a new series of stable and well-defined NIR-absorbing metallacycles (M1-M3) through the Pt-acetylide coordination with highly efficient photoconversion performance for cancer phototherapy. The metallacycles showed high stability and strong NIR absorption, and the absorption peaks were red shifted approximately 30 nm in comparison with their corresponding precursors. The introduction of Pt into metallacycles promotes significant photoconversions, including the singlet-to-triplet and nonradiative transitions. Moreover, the fabricated M3 nanoparticles (M3-NPs) showed favorable photoconversions into both thermal effect and singlet oxygen generation upon NIR irradiation, achieving tumor ablation. This novel design of Pt-acetylide metallacycles possesses not only complex topological architectures but also a valuable paradigm for precise cancer phototherapy, which is important for grafting stimuli-responsive functional groups into metallacycles for the development of high-performance biomedical supramolecular materials.

Pure white light emission and charge transfer in organogels of symmetrical and unsymmetrical π-chromophoric oligo-p-(phenyleneethynylene) bola-amphiphiles.

Two novel bola-amphiphilic oligo-p-(phenyleneethynylene) (OPE) dicarboxylates have been synthesized having non-polar and mixed-polar side chains. This led to gelation in both with vesicular morphology. Upon in situ loading of a suitable dye and redox-active molecule, pure white light emitting and charge transfer (CT)-gels, respectively, were realized.

Synthesis of carbonylated heteroaromatic compounds via visible-light-driven intramolecular decarboxylative cyclization of o-alkynylated carboxylic acids.

An efficient strategy for the easy access to carbonylated heteroaromatic compounds has been developed via a visible-light-promoted intramolecular decarboxylative cyclization reaction of o-alkynylated carboxylic acids. This method is characterized by its benign conditions and the tolerance to a wide range of functionalities.

In vitro cytotoxicity of antimicrobial conjugated electrolytes: interactions with mammalian cells.

An estimated 19 000 deaths and $3-4 billion in health care costs per year in the United States are attributed to methicillin-resistant Staphlococcus aureus (MRSA) infections. Certain conjugated phenylene ethynylene (CPE)-based polymers (PPE) and oligomers (OPE) have been demonstrated to exhibit dark and light-activated antimicrobial activity. Until recently, the relative cytotoxicity of these PPEs and OPEs toward mammalian cells haas been unknown, limiting the applications for which they may be used (e.g., reducing and/or preventing the spread of untreatable bacterial strains). In this work, we examine the toxicity of CPEs to mammalian cells using cytotoxicity assays of cellular monolayers. Eight CPEs, two PPEs and six OPEs, were selected for these studies based on their biocidal activity, and diversity of repeat unit number and functional groups. Briefly, two cell types were exposed to CPEs at concentrations ranging from 1-100 ug/mL for 24 h. We find that concentration largely determines the resulting viability of cells, although at intermediate concentrations (5-10 ug/mL), the effect of light on light-activated compounds is very important. Furthermore, we find that the longer-chained compounds are cytotoxic at much higher concentrations, and therefore have the widest range of concentrations available for potential applications.

The effect of hydrogen-bonds of amino acid-derived diacetylene by photopolymerization in supramolecular hydrogels.

Amino acids derived diacetylene hydrogelators 1 and 2 which have glycine or alanine moiety as head group were synthesized by several steps. The influences on the hydrogen-bonding interaction between amino acids moieties of hydrogelators in the polymerization by UV irradiation were investigated. The glycine-based hydrogel 1 resulted in the generation of stronger absorption peak at 540 nm at room temperature by UV-irradiation. Interestingly, the red color of the glycine-based diacetylene hydrogel 1 was gradually changed into blue color by decreasing temperature. Finally, the absorption band with 640 nm of glycine-based diacetylene hydrogel 1 showed at 77 K. On the other hand, the alanine-based hydrogel 2 had no significant color change at 77 K. These different color changes between the hydrogels 1 and 2 are attributed to the strong intermolecular hydrogen-bonding interaction between glycine moieties of the hydrogelator 1, as confirmed by FTIR observation. These results indicate that the hydrogen-bonding strength of the self-assembled hydrogel is a strong influence on the degree of polymerization.

Light-induced antibacterial activity of symmetrical and asymmetrical oligophenylene ethynylenes.

The light-induced antibacterial activity of symmetric and asymmetric oligophenylene ethynylenes (OPEs) was investigated against Gram-positive (Staphylococcus aureus and Staphylococcus epidermidis) and Gram-negative (Escherichia coli) bacteria. To understand the light-induced biocidal effect better, the transient absorption and triplet lifetime of OPEs were studied in methanol and water. A higher triplet lifetime was observed for OPE samples in water than in methanol. The magnitudes of the changes in optical density (ΔOD) of the S-OPE-n(H) series of symmetric oligomers are much higher than that of the asymmetric OPE-n series in water and are generally correlated with the singlet oxygen yield. It was found that the antibacterial activity against both Gram-positive and Gram-negative bacteria is size-, concentration-, and time-dependent. The light-induced antibacterial activity may result from the coordinated interactions of membrane disruption and interfacial or intracellular singlet oxygen generation, and the dominant factor is most likely the latter. The results obtained in this study will aid in the design of more efficient biocides in the future.

Synthesis of glycoluril catalyzed by potassium hydroxide under ultrasound irradiation.

Synthesis of the glycolurils catalyzed by potassium hydroxide was carried out in 17-75% yield at 40 degrees C in EtOH under ultrasound irradiation. Compared to the method using stirring, the main advantage of the present procedure is milder conditions and shorter reaction time.

Stereocontrolled synthesis of carbocycles via four successive pericyclic reactions.

Herein, we report a highly diastereoselective construction of fused carbocycles using four successive pericyclic reactions.

Singlet and triplet excited-state interactions and photochemical reactivity of phenyleneethynylene oligomers.

The rigid rodlike character of phenyleneethynylenes and their ability to communicate charge/excitation energy over long distances have made them useful as molecular linkers in the light energy harvesting assemblies and molecular electronics devices. These linker molecules themselves possess rich photochemistry as evident from the relatively large yields of the excited singlet (0.5-0.66) and triplet (0.4-0.5) states of two model oligomers, 1,4-bis(phenylethynyl)-2,5-bis(hexyloxy)benzene (OPE-1) and 1,4-bis((4-phenylethynyl)phenylethynyl)-2,5-bis(hexyloxy)benzene (OPE-2). In particular, the long-lived triplet excited state is capable of undergoing deactivation by self-quenching processes such as ground-state quenching and triplet-triplet (T-T) annihilation. The T-T annihilation occurs with a nearly diffusion-controlled rate (approximately 2 x 10(9) M(-1) s(-1)), and ground-state quenching occurs with a rate constant of approximately 6 x 10(7) M(-1) s(-1). The electron transfer from the excited OPE-1 and OPE-2 to benzoquinone as characterized from the transient absorption spectroscopy illustrates the ability of these molecules to shuttle the electrons to acceptor moieties. In addition, pulse radiolysis experiments confirm the spectroscopic fingerprint of the cation radical (or "trapped hole") with absorption bands in the 500-600 nm region.

Synthesis and protein degradation capacity of photoactivated enediynes.

[structure: see text] The viability of proteins as targets of thermally and photoactivated enediynes has been confirmed at the molecular level. Model studies using a labeled substrate confirmed the efficacy of atom transfer from diyl radicals produced from enediynes to form captodatively stabilized carbon centered aminoacyl radicals, which then undergo either fragmentation or dimerization. To exploit this finding, a family of enediynes was developed using an intramolecular coupling strategy. Derivatives were prepared and used to target specific proteins, showing good correlation between affinity and photoinduced protein degrading activity. The findings have potential applications in the design of artificial chemical proteases and add to our understanding of the mechanism of action of the clinically important enediyne antitumor antibiotics.

Preparation of bis(diazo) compounds incorporated into butadiyne and thiophene units and generation and characterization of their photoproducts.

(2,6-dimethyl-4-tert-butylphenyl)(2,4,6-tribromophenyl)diazomethane(-N(2)) was found to be stable enough to survive under Sonogashira coupling reaction conditions, and aryldiazomethyl substituents were introduced at the 1,4-positions of butadiyne (4-2N(2)) and the 2,5-positions of thiophene(5-2N(2)). Irradiation of those bis(diazo) compounds generated bis(carbenes), which were characterized by using ESR and UV/vis spectroscopic techniques in a matrix at low temperature as well as time-resolved UV/vis spectroscopy in solution at room temperature. These studies revealed that both of the bis(carbenes), 4 and 5, have singlet quinoidal diradical ground states with a very small singlet-triplet energy gap of less than 1 kcal mol(-1). A remarkable increase in the lifetime of bis(carbenes), as opposed to that of the monocarbene (2), was noted and was interpreted to indicate that bis(carbenes) are thermodynamically stabilized as a result of delocalization of unpaired electrons throughout the pi net framework. In spite of the stability, both bis(carbenes) are readily trapped by molecular oxygen to afford bis(ketones). Presumably, the reaction of the upper-lying localized quintet states with oxygen is much faster than that for lower-lying states.

[Mixed monolayers of amphiphile-modified nucleic bases and diynoic acids. I. Phase states at air-water interface and in Langmuir-Blodgett films]. / Smeshannye monosloi amfifil'nykh proizvodnykh azotistykh osnovanii i diinovykh kislot. I. Fazovoe sostoianie na granitse voda-vozdukh i v plenkakh Lengmiura-Blodzhett.

Monolayers of amphiphile-modified nucleic bases with diynoic acid were obtained and characterized. The synthesized nucleic bases contained in the monolayer complementarily bind the nucleotide molecules contained in the aqueous subphase, and the structure of the resulting monolayers can be fixed by the photopolymerization of diynoic acid. The resulting monolayer exemplifies a novel type of model systems for investigating molecular recognition at the surface of biological membranes. Procedures for the transfer of the monolayers onto solid substrates and photopolymerization of the diynoic acid in mixtures with the derivatives of nucleic bases were developed. The films obtained were structurally characterized using atomic force microscopy. Compression isotherms of the mixed monolayers as well as individual components of monolayers at the air-water interface allowed one to determine the concentration range at which the diynoic acid form true mixtures or domain structures with the derivatives of nucleic base. A study of the films transferred to the solid substrate by atomic force microscopy indicated that this concentration dependence of miscibility behavior was conserved in the transferred films.

Synthesis and DNA damaging ability of enediyne model compounds possessing photo-triggering devices.

Enediyne model compounds possessing photo-triggering devices were developed. These enediynes afforded biradicals by UV irradiation and showed DNA cleaving activity. The DNA damage was confirmed to be mainly caused by the biradical, not singlet oxygen.

A novel concept to activate enediynes for DNA cleavage.

A novel concept is presented to activate enediynes via a biscumulenic intermediate using photoinduced electron transfer (PET).

The ovicidal activity of cis-dehydromatricaria ester: time-dependance of its enhancement by UV light.

Freshly laid eggs of Drosophila melanogaster were treated with the polyacetylenic compound cis-dehydromatricaria ester. The toxicity of the chemical was enhanced by treatments with long-wavelength ultraviolet light, and the maximum effect was observed when the UV irradiation was performed 4-5 h after the initial contact of the eggs with the chemical.

Ultraviolet-mediated antibiotic activity of species of Compositae caused by polyacetylenic compounds.

A survey of North, Central, and South American composites, and of certain polyacetylenic compounds occurring in them, confirmed that the ultraviolet-mediated antibiotic activity against Candida albicans can be ascribed to the presence of particular polyacetylenes and their thiophene derivatives. Leaves, stems, roots, and achenes were assayed separately. An attempt was made to relate the phototoxic activity of specific compounds to their chemical structures. Most composites tested were not phototoxic against Candida, but many were antibiotic. The antibiotic activity also appears to be caused by polyacetylenes. Of 65 sequiterpene lactones assayed, only a few were found to be antibiotic, and only one, glaucolide G, was phototoxic.