Selective Detection of Choline in Pseudophysiological Medium with a Fluorescent Cage Receptor.

A fluorescent cage receptor for the detection of choline in pseudophysiological medium is described. Not only does this capsule complex choline with an association constant greater than 9.9 × 104 M-1 in buffered medium but it is also selective toward acetylcholine.

Hemicryptophanes with Improved Fluorescent Properties for the Selective Recognition of Acetylcholine over Choline.

The synthesis of two new fluorescent hemicryptophanes is reported. They were found to be efficient and selective receptors for acetylcholine over choline. When compared to other hemicryptophane hosts previously reported for the selective recognition of acetylcholine, they display improved fluorescent properties: their maximum emission wavelengths are red-shifted and the quantum yields are higher. NMR titration experiments and density functional theory (DFT) calculations support the results obtained from fluorescence spectroscopy and give insights into the interactions involved in the host/guest complexes and into the selectivity for acetylcholine over choline.

Selective recognition of acetylcholine over choline by a fluorescent cage.

A fluorescent hemicryptophane has been synthesized and can be used as a turn on receptor of acetylcholine. A binding constant of 2.4 × 104 M-1 was measured for this neurotransmitter, and its selective and sensitive detection over choline and choline phosphate was achieved. NMR and DFT calculations provide insight into the interactions involved in this selective recognition process.

Synthesis and Structural Studies of Gallium(III) and Iron(III) Hemicryptophane Complexes.

New gallium(III) and iron(III) endohedral complexes were obtained from a hemicryptophane ligand bearing suitable binding sites for octahedral metal coordination. The solid-state structures of the free host and of the complexes were determined by single-crystal X-ray diffraction analysis. The metal ion is linked to the hydrazone nitrogen and the phenolate oxygen atoms, yielding a distorted octahedral geometry around the encapsulated metal. The two isomorphous structures of the metal complexes reveal the exclusive formation of PΔ/MΛ enantiomeric pairs.

Differential Photoluminescent and Electrochemiluminescent Behavior for Resonance Energy Transfer Processes in Thermoresponsive Microgels.

Stimuli-responsive microgels with redox and luminescent resonance energy transfer (LRET) properties are reported. Poly(N-isopropylacrylamide) microgels are functionalized simultaneously with two models dyes: a derivative of tris(bipyridine) ruthenium complex and cyanine 5. Both moieties are chosen as a pair of luminophores with a spectral overlap for resonance energy transfer, where the ruthenium complex acts as a donor and the cyanine an acceptor. The effect of the temperature on the efficiency of the LRET of the microgels has been investigated and compared using either photoluminescence (PL) or electrochemiluminescence (ECL) as the excitation process. In PL, the synthesized microgels exhibit resonance energy transfer regardless of the swelling degree of the microgels. The transfer efficiency is a function of the donor-acceptor distance and can be tuned either by the swell-collapse phase transition or by the dye content in the microgel network. In ECL, the microgels emit light only at the wavelength of the ruthenium complex because the resonance energy transfer does not occur. Indeed, even within the microgel matrix, the cyanine dye is oxidized at the potential required for ECL generation, which impairs its emitting properties. Thus, both excitation channels (i.e., PL and ECL) show differential behavior for the resonance energy transfer processes.

Selective, direct detection of acetylcholine in PBS solution, with self-assembled fluorescent nano-particles: experiment and modelling.

We report synthesis, characterisation and molecular modelling of a new fluorescent cyclotriveratrylene probe for acetylcholine in aqueous media, with emission around 430 nm thanks to extended conjugation. The probe discriminates acetylcholine from choline, with respective binding constants 540 and 240 M(-1) in PBS buffered saline solution, an order of magnitude improvement over the previous best performance. Dynamic light scattering and transmission electron microscopy show the new probe self-assembles in ca. 5 nm diameter particles in PBS medium. Molecular modelling suggests that the high fluorescence quantum yield of the probe, 20% in aqueous media, is due to features of the molecular arrangement in the nano-particles, contributing both to exposure of the complexation site and to shielding of the fluorescent π system from quenching by water. Titration data for other quaternary ammoniums and modelling indicate that recognition of acetylcholine vs. choline depends on specific electrostatic interactions, and to a lesser extent on exclusion of water by hydrophobic-hydrophilic segregation. Probe-substrate interactions enhance the fluorescence of the probe by shielding against water and by flattening the π system.

Enhanced electrogenerated chemiluminescence in thermoresponsive microgels.

The electrochemistry, photoluminescence and electrogenerated chemiluminescence of thermoresponsive redox microgels were investigated. For the first time, reversible ECL enhancement is demonstrated in stimuli-responsive 100-nm microgel particles. Such an unexpected amplification reached 2 orders of magnitude, and it is intrinsically correlated with the collapse of the microgel particles. The swell-collapse transition decreases the average distance between adjacent redox sites and favors the electron-transfer processes in the microgels resulting in the enhanced ECL emission.

Electrochemistry and bioactivity relationship of 6-substituted-4H-pyrido[4,3,2-kl]acridin-4-one antitumor drug candidates.

We report here the electrochemical characterization of eight synthetic DNA intercalators based on the 4H-pyrido[4,3,2-kl]acridin-4-one structure. We found that the electrochemical behavior of these redox active drugs is strongly influenced by the nature of the solvent. A single two-electron reduction is observed in an aqueous phosphate buffer (PB) whereas two successive one-electron reductions are observed in aprotic solution (acetonitrile). The influence of the molecular structure on the potential values is addressed along with a comparison between the DNA binding constant (K(DNA)) and the cytotoxic activity against HT29 cells (IC(50)). For typical DNA intercalators, one could expect that toxicity will be roughly proportional to the DNA binding constant. Yet, a structure/activity comparison solely based on the DNA affinity was not conclusive. In contrast, a direct relationship was evidenced for the first time between the decimal logarithm of the in vitro bioactivity and the reduction potential of pyridoacridones recorded in PB at pH 7.0. Moreover, most of the bio/electrochemical relationships previously described for quinone-based drugs were reported with electrochemical characterization in aprotic solvents (typically acetonitrile, dimethylformamide or dimethylsulfoxide). But aqueous solution electrochemistry is definitely the most bio-relevant because the redox mechanism of quinone or iminoquinone reduction directly depends on the protic nature of the solvent.

Development of functionalized cyclotriveratrylene analogues: introduction of withdrawing and π-conjugated groups.

Cyclotriveratrylene analogues (CTVs) are supramolecular bowl-shaped molecules known for their ability to complex organic and organometallic guests, to form liquid crystals, polymers, or nanostructures. In this Article, we report the synthesis of new cyclotriveratrylene analogues with fluorescence properties in which various electron-withdrawing or π-extended conjugated groups are appended to the wide rim ortho to the methoxy-donating groups. Synthetically, these functionalized CTVs cannot be obtained as CTVs with electron-rich functions by the typical method (i.e., the trimerization of the corresponding benzyl alcohol) but are prepared from a common key intermediate, the C(3)-triiodocyclotriveratrylene (CTV-I(3)), in good yields. Despite the synthetic difficulties encountered due to the presence of three reactive centers, we have demonstrated the possibility of performing Sonogashira coupling and Huisgen cycloaddition reactions directly to the CTV core for the first time. CTVs with π-extended conjugated groups reveal interesting fluorescence profiles. More broadly, this study utilizes CTV-I(3) to introduce novel functionalities into CTVs to keep exploring their potential applications.

The fraction of α-linolenic acid present in the sn-2 position of structured triacylglycerols decreases in lymph chylomicrons and plasma triacylglycerols during the course of lipid absorption in rats.

Little is known about the ability of α-linolenic acid (Ln) to remain in the sn-2 position of TG during the absorption process. The goal of this study was to determine the Ln distribution in the lymph (Study 1) and plasma (Study 2) TG of rats fed a single i.g. load of structured TG [300 mg/rat of either oleic acid (O)/Ln/O TG (OLnO) or Ln/O/O TG (LnOO), n = 7 rats]. In an early fraction (3-4 h) of lymph (OLnO group; 100% Ln in the sn-2 position), 46 ± 2% Ln was maintained in this position in lymph TG. There was even less (29 ± 6%) in the last fraction (7-24 h) (P < 0.05). Ln was also found (9 ± 3%) in the sn-2 position of lymph TG in the LnOO group. The Ln content in lymph phospholipids was twice as high in rats when they were fed LnOO (4.2 ± 0.1%) than OLnO (2.3 ± 0.2%) (P < 0.005). Six hours postprandially (Study 2), 21 ± 3% of the Ln incorporated into plasma TG was located in the sn-2 position in the OLnO group compared to 13 ± 2% in the LnOO group (P < 0.001). Overall, these results indicate that the amount of Ln that moved from the sn-2 position of structured TG to the sn-1(3) position of lymph TG increased during absorption. This may account for a substantial hydrolysis of the 2-monolinolenylglycerols in enterocytes, leading to the intramolecular redistribution of Ln in lymph TG and, consequently, in plasma TG.

C3-triiodocyclotriveratrylene as a key intermediate to fluorescent probes: application to selective choline recognition.

A new strategy to obtain fluorescent cyclotriveratrylene (CTV) probes is proposed. The key intermediate, a triiodo CTV, is prepared in 3 steps with 47% overall yield. The whole synthesis requires only one purification step. The potential of this triiodo CTV as an intermediate is illustrated through the synthesis of a fluorescent phosphorylated probe that is able to bind choline and acetylcholine in pseudo-physiological conditions, with selectivity towards choline. As a consequence, this intermediate should allow us to rapidly form a library of probes in order to highlight the most promising ones.

Designed glucose-responsive microgels with selective shrinking behavior.

We report on the synthesis of various glucose-responsive microgels based on N-alkylacrylamide derivatives and phenylboronic acid (PBA) as a glucose sensing moiety. Depending on their chemical composition, the microgels exhibit opposite behaviors in response to glucose concentration increase: they can either swell or shrink, using two different mechanisms for glucose recognition. Both behaviors may be suitable for glucose sensing and insulin delivery. When glucose binds a single boronate receptor, the microgel swells as glucose concentration increases. This mechanism can be used to deliver a drug by diffusion through the network. In other cases, glucose binds specifically to two boronates, which creates additional cross-links within the network and provokes shrinkage. Such systems are promising for the development of sensors with improved selectivity and also as potential "intelligent" valves in microfabricated delivery systems. By a rational choice of the constituting units of the network structure, we show how to favor one or the other type of response to glucose variation. Therefore, glucose-swelling microgels operating under physiological conditions have been obtained by copolymerization with an appropriate choice of alkylacrylamide monomer and boronate derivative. At a pH above the pK(a) of the boronic acid derivative, the same structures shrink in response to glucose concentration. The nature of the cross-linker is a key parameter to enable this dual behavior. In other microgels, an amine group is introduced in the vicinity of the boronic acid, which lowers its pK(a) and favors microgel contraction at physiological pH. This work has allowed us to give some general rules to control the swelling/shrinking behavior of glucose-responsive microgels.

A fluorescent cyclotriveratrylene: synthesis, emission properties and acetylcholine recognition in water.

A fluorescent cyclotriveratrylene 1 was synthesized and characterized in methanol and water. Soluble in pure water and physiological media, compound 1 has binding properties towards acetylcholine. This detection is direct, contrary to most fluorescent systems which rely upon a competition principle between the guest and a fluorophore.

Differential photoluminescent and electrochemiluminescent detection of anions with a modified ruthenium(II)-bipyridyl complex.

A new guanidinium 3,3'-functionalized bipyridylruthenium(II) complex has been prepared for the differential sensing of L-glutamate and dihydrogenphosphate anions depending on the luminescent detection scheme. The effects of such anions on the photoluminescent (PL) and electrochemiluminescent (ECL) properties of the complex have been investigated and compared. The PL intensity increases up to fourfold in the presence of L-glutamate. The increase of intensity in the presence of dihydrogenphosphate is weaker and no change in PL intensity is observed in presence of acetate, iodide, or chloride anions. With n-tripropylamine, ECL emission of the Ru(II) complex is initiated at 1.45 V versus Ag/AgCl/KCl and the ECL intensity increases only in the presence of dihydrogenphosphate. Indeed, L-glutamate is already oxidized at the relatively high potential required for ECL generation and thus it does not affect the ECL signal. The comparison of the competitive ECL and PL assays in a mixture of anions confirms the differential detection of L-glutamate and of dihydrogenphosphate. Thus, both sensing channels (i.e., PL and ECL) show different selectivities depending on the nature and on the electroactivity of the target anions. Multianion analysis is demonstrated in competitive assays using complementary detection methods.

Monodispersed glucose-responsive microgels operating at physiological salinity.

Monodispersed poly(N-isopropylacrylamide) submicrometric microgels modified with a phenylboronic acid (PBA) derivative have been synthesized by precipitation polymerization. Particles with a well-controlled size and adjustable composition were obtained. These particles were found to be glucose responsive at a pH close to the pKa of the PBA derivative, with a swelling degree proportional to the concentration of glucose. In addition, the response to glucose was found to strongly depend on the initial state of the microgel, which depended itself on the initial temperature and the functionalization degree of the particle. This result explained the fundamental difference in the behavior of PBA-poor particles and rich ones in the presence of electrolyte. Interestingly, the latter exhibited a high swelling ratio in the presence of glucose at physiological electrolyte concentration. These particles may serve as building blocks for the design of colorimetric sensors based on the light diffraction of colloidal crystals.

Photoswitching of cation complexation with A monoaza-crown dithienylethene photochrome.

A photochromic dithienylethene, bearing a phenyl azacrown as an ionophore and a formyl group as an electron-accepting substituent, changes its binding ability for Ca2+ by a factor higher than 103 by photoirradiation. This new photoionochromic displays a wavelength-dependent competition between fluorescence and photocyclization assigned to a red-shifted absorption of the fluorescing conformer compared to the absorption of the photoreactive conformer.