Studies About the Effect of Halogenated Solvents on the Fluorescence Properties of 9-Aryl-Substituted Isoquinolinium Derivatives - A Case Study.

It was demonstrated that 9-aryl-substituted isoquinolinium derivatives have significantly increased fluorescence quantum yields in halogenated solvents, mostly pronounced in chloroalkanes, which appears to be specific for this type of solvents. Further analysis with selected halogenated solvents revealed that the type and number of halogen substituents and the dielectric constant of the solvent have a distinct impact on the emission quantum yield. The solvent effect is explained by a solvation of the charge shift (CS) state by attractive halogen-π interactions (halogen bond), which impedes the torsional relaxation of the excited state.

Photocontrolled Binding of Styrylnaphthyridine Ligands to Abasic Site-Containing DNA by Reversible [2+2] Cycloaddition and Cycloreversion.

Five novel styrylnaphthyridine derivatives were synthesized and shown to operate as photoswitchable, selective ligands for abasic site-containing DNA (AP-DNA), which is an important therapeutic and diagnostic target. These compounds associate with AP-DNA with binding constants of 0.5-8.4×104 M-1 as shown by photometric and fluorimetric titrations. Specifically, these ligands bind preferentially to AP-DNA relative to regularly paired duplex DNA. As a special feature, the association of these ligands with DNA can be controlled by means of a reversible [2+2] photocycloaddition. Upon irradiation at 420 nm the photodimer is formed, which does not bind to AP-DNA. In turn, the naphthyridine is regained with excitation at 315 nm. Most notably, this photoinduced deactivation and release of the DNA ligand can be performed in situ in the presence of AP-DNA, thus providing a tool for on-demand delivery of a DNA binder. Overall, these results provide a promising starting point for the development of functional AP-DNA ligands whose bioactivity can be modulated by light with local and temporal control.

Reversible Restrain and Release of the Dynamic Valence Isomerization in a Shape-shifting Bullvalene by Complex Formation.

In search for structural features that enable the control of the valence isomerization of the fluxional bullvalene, a bullvalene-bis(harmane) conjugate is identified that acts as chelating ligand in complexes with metal ions. Spectrometric titrations show that this ligand forms 1 : 1 complexes with Ag+, Cu+, Cu2+, and Zn2+. Most importantly, detailed NMR-spectroscopic analysis at different temperatures reveals that the complexation with Ag+ strongly affects the dynamic isomerization of the bullvalene unit of the ligand such that only one predominant valence isomer is formed, even at 5 °C. Detailed 1H-NMR-spectroscopic studies disclose an increased barrier (~11 kJ mol-1) of the Cope rearrangement. Furthermore, the addition of hexacyclene displaces the Ag+ from the complex, so that the valence isomerization is accelerated and an equilibrium with two predominant isomers is formed. In turn, repeated addition of Ag+ regains the complex with the restrained isomerization of the bullvalene unit. This method to control the valence isomerism by straightforward chemical stimuli may be used to simplify structural analysis at elevated temperatures, i. e. a feature not available so far with bullvalenes, and it may be employed as functional element in dynamic supramolecular assemblies.

Photoinduced in situ generation of DNA-targeting ligands: DNA-binding and DNA-photodamaging properties of benzo[c]quinolizinium ions.

The photoreactions of selected styrylpyridine derivatives to the corresponding benzo[c]quinolizinium ions are described. It is shown that these reactions are more efficient in aqueous solution (97-44%) than in organic solvents (78-20% in MeCN). The quinolizinium derivatives bind to DNA by intercalation with binding constants of 6-11 × 104 M-1, as shown by photometric and fluorimetric titrations as well as by CD- and LD-spectroscopic analyses. These ligand-DNA complexes can also be established in situ upon irradiation of the styrylpyridines and formation of the intercalator directly in the presence of DNA. In addition to the DNA-binding properties, the tested benzo[c]quinolizinium derivatives also operate as photosensitizers, which induce DNA damage at relative low concentrations and short irradiation times, even under anaerobic conditions. Investigations of the mechanism of the DNA damage revealed the involvement of intermediate hydroxyl radicals and C-centered radicals. Under aerobic conditions, singlet oxygen only contributes to marginal extent to the DNA damage.

Fluorimetric Cell Analysis with 9-Aryl-Substituted Berberine Derivatives as DNA-Targeting Fluorescent Probes.

DNA-sensitive fluorescent light-up probes based on berberine are presented. This biogenic fluorophore was chosen as central unit to use its potential biocompatibility and its DNA-binding properties. To provide predictable fluorescence quenching in aqueous solution and a fluorescence light-up effect upon DNA binding, aryl substituents were attached at the 9-position by Suzuki-Miyaura coupling reactions. The 9-arylberberine derivatives have a very low fluorescence quantum yield (Φfl =<0.02), which is caused by the radiationless deactivation of the excited state by torsional relaxation about the biaryl axis. In addition, these berberine derivatives intercalate into DNA with high affinity (Kb =2.0-22×104  M-1 ). Except for the nitrophenyl- and hydroxyphenyl-substituted derivatives, all tested compounds exhibited a pronounced fluorescence light-up effect upon association with DNA, because the deactivation of the excited-state by torsional relaxation is suppressed in the DNA binding site. Most notably, it was shown exemplarily with the 9-(4-methoxyphenyl)- and the 9-(6-methoxynaphthyl)-substituted derivatives that these properties are suited for fluorimetric cell analysis. In particular, these probes generated distinct staining patterns in eukaryotic cells (NIH 3T3 mouse fibroblasts), which enabled the identification of nuclear substructures, most likely heterochromatin or nucleoli, respectively.

Bis- and Tris-norbornadienes with High Energy Densities for Efficient Molecular Solar Thermal Energy Storage.

Molecular solar thermal energy storage (MOST) systems can convert, store and release solar energy in chemical bonds, i.e., as chemical energy. In this work, phenyl- and naphthyl-linked bis- and tris-norbornadienes are presented as promising MOST systems with very high energy densities. The substrates were synthesized by Suzuki-Miyaura coupling reactions and their absorption properties and characteristic parameters for MOST applications were investigated. The norbornadiene derivatives showed absorption onsets of up to 386 nm and photoisomerization quantum yields of 56 % per photoisomerization event. The resulting quadricyclane products have half-lifes up to 14 d and very high energy densities of up to 734 kJ/kg. Overall, these norbornadienes fulfill necessary criteria for an optimal MOST system and are, therefore, a highly promising basis for the development of materials for efficient solar energy conversion and storage.

Switching between DNA binding modes with a photo- and redox-active DNA-targeting ligand, part II: the influence of the substitution pattern.

A disulfide-functionalized photoactive DNA ligand is presented that enables the control of its DNA-binding properties by a combination of a photocycloaddition reaction and the redox reactivity of the sulfide/disulfide functionalities. In particular, the initially applied ligand binds to DNA by a combination of intercalation and groove-binding of separate benzo[b]quinolizinium units. The association to DNA is interrupted by an intramolecular [4 + 4] photocycloaddition to the non-binding head-to-head cyclomers. In turn, the subsequent cleavage of these cyclomers with dithiothreitol (DTT) regains temporarily a DNA-intercalating benzoquinolizinium ligand that is eventually converted into a non-binding benzothiophene. As a special feature, this sequence of controlled deactivation, recovery and internal shut-off of DNA-binding properties can be performed directly in the presence of DNA.

Switching between DNA binding modes with a photo- and redox-active DNA-targeting ligand.

A disulfide-functionalized bis-benzo[b]quinolizinium is presented that is transformed quantitatively into its cyclomers in a fast intramolecular [4 + 4] photocycloaddition. Both the bis-quinolizinium and the photocyclomers react with glutathione (GSH) or dithiothreitol (DTT) to give 9-(sulfanylmethyl)benzo[b]quinolizinium as the only product. As all components of this reaction sequence have different DNA-binding properties, it enables the external control and switching of DNA association. Hence, the bis-benzo[b]quinolizinium binds strongly to DNA and is deactivated upon photocycloaddition to the non-binding cyclomers. In turn, the subsequent cleavage of the cyclomers with DTT regains a DNA-intercalating benzoquinolizinium ligand. Notably, this sequence of controlled deactivation and recovery of DNA-binding properties can be performed directly in the presence of DNA.

Ligands for Abasic Site-containing DNA and their Use as Fluorescent Probes.

Apurinic and apyrimidinic sites, also referred to as abasic or AP sites, are residues of duplex DNA in which one DNA base is removed from a Watson-Crick base pair. They are formed during the enzymatic repair of DNA and offer binding sites for a variety of guest molecules. Specifically, the AP site may bind an appropriate ligand as a substitute for the missing nucleic base, thus stabilizing the abasic site-containing DNA (AP-DNA). Notably, ligands that bind selectively to abasic sites may be employed for analytical and therapeutical purposes. As a result, there is a search for structural features that establish a strong and selective association of a given ligand with the abasic position in DNA. Against this background, this review provides an overview of the different classes of ligands for abasic site-containing DNA (AP-DNA). This review covers covalently binding substrates, namely amine and oxyamine derivatives, as well as ligands that bind to AP-DNA by noncovalent association, as represented by small heterocyclic aromatic compounds, metal-organic complexes, macrocyclic cyclophanes, and intercalator-nucleobase conjugates. As the systematic development of fluorescent probes for AP-DNA has been somewhat neglected so far, this review article contains a survey of the available reports on the fluorimetric response of the ligand upon binding to the AP-DNA. Based on these data, this compilation shall present a perspective for future developments of fluorescent probes for AP-DNA.

Pushing Photochemistry into Water: Acceleration of the Di-π-Methane Rearrangement and the Paternó-Büchi Reaction "On-Water".

Two representative organic photoreactions, namely a bimolecular photocycloaddition and a monomolecular photorearrangement, are presented that are accelerated when the reaction is performed "on-water", that is, at the water-substrate interface with no solvation of the reaction components. According to the established models of ground-state reactions "on-water", the enhanced efficiency of the photoreactions is explained by hydrophobic effects (Paternó-Büchi reaction) or specific hydrogen bonding (di-π-methane rearrangement) at the water-substrate interface that decrease the energy of the respective transition state. These results point to the potential of this approach to conduct photoreactions more efficiently in an ecologically favorable medium.

The Stereoselective Total Synthesis of Axially Chiral Naphthylisoquinoline Alkaloids.

The naphthylisoquinoline (NIQ) alkaloids are a thrilling class of natural biaryls─structurally, biosynthetically, and pharmacologically. A common feature of these metabolites is the biaryl bond between their naphthalene and isoquinoline moieties, which in most cases is rotationally hindered, leading to the phenomenon of axial chirality. Depending on their individual structures, including the respective axial configurations, NIQs show promising bioactivities. Their total synthesis is a challenging but rewarding goal, with the stereocontrolled construction of the biaryl linkage as the key step.The position of the biaryl axis and its configuration determine the overall molecular shape and thus the choice of the best possible method for efficient asymmetric aryl-aryl bond formation. The axis in NIQs can cover a broad range of steric hindrance, from freely rotating to configurationally stable. For dioncophylline B (1) and dioncophylline F (2a/b), with only two ortho-substituents next to the axis, the synthesis is easy to accomplish by direct coupling of the intact naphthalene moiety with the isoquinoline unit, and no atropo-selectivity is required.Naphthylisoquinolines with a configurationally stable biaryl axis are the focus of the present Account. They are more difficult to synthesize because, in addition to the problem of decreased chemical yields with increasing steric hindrance at the axis, the synthesis needs to proceed stereoselectively. Within this class of NIQs, 5,8'-coupled representatives, such as korupensamine A (3a), have received considerable synthetic attention because the rotational barrier is high enough for the existence of atropisomerism without being too excessive, and they show potent bioactivities. Their synthesis, as systematically presented herein, thus occupies a central role in this report. For their aryl-aryl bond formation, both intra- and intermolecular approaches can be successfully applied. Axial stereoinformation is introduced by internal asymmetric induction from stereogenic elements already present in the isoquinoline or its precursors, from chiral auxiliary elements artificially introduced, or by external asymmetric induction using chiral catalysts.To overcome even higher steric hindrance, as in ancistrocladine (4a), innovative approaches were developed. A most successful strategy is the "lactone concept" developed by the Bringmann group, which allows the directed synthesis of any desired atropisomer in high chemical and optical yields, thus permitting the atropo-divergent preparation of the two isomers from a single joint precursor. In this approach, the two formal tasks of stereoselective biaryl synthesis, which are usually done simultaneously─the C-C linkage and the asymmetric induction─are achieved consecutively. The coupling step is performed intramolecularly after prefixation of the coupling partners by an ester bridge. The resulting biaryl lactone already possesses the biaryl axis but is still configurationally unstable; it can then, with internal or external asymmetric induction, be cleaved atropo-divergently with high stereoselectivities. Besides its unique concept, the procedure excels by its broad applicability; among all presented methods, it has been used for the synthesis of the largest number of NIQs, more than 20 representatives, including those with the highest steric hindrance.This Account gives comprehensive insight into the plethora of conceptual approaches for the efficient formation of the hindered biaryl bond of NIQs.

Borylated norbornadiene derivatives: Synthesis and application in Pd-catalyzed Suzuki-Miyaura coupling reactions.

The photochromic norbornadiene/quadricyclane system is among the most promising candidates for molecular solar thermal (MOST) energy storage. As in this context there is still the need for new tailor-made derivatives, borylated norbornadienes were synthesized that may be used as versatile building blocks. Thus, the 4,4,5,5-tetramethyl-2-(bicyclo[2.2.1]heptadien-2-yl)-1,3,2-dioxaborolane was prepared and shown to be a suitable substrate for Pd-catalyzed Suzuki-Miyaura coupling reactions with selected haloarenes. It was demonstrated exemplarily that the novel monosubstituted 2-(1-naphthyl)norbornadiene, that is accessible through this route, is transformed to the corresponding quadricyclane upon irradiation, whereas the back reaction can be accomplished by thermal treatment.

Synthesis of Fluorescent, DNA-Binding Benzo[b]indolonaphthyridinium Derivatives by a Misguided Westphal Condensation.

A novel type of azoniahetarene, namely, benzo[b]indolonaphthyridinium, was unexpectedly formed by the reaction of N-alkylated ß-carbolinium derivatives and the enolizable 1,2-cyclohexadione under typical conditions of a Westphal reaction. The products exhibit high fluorescence intensities in polar solvents (Φfl = 0.52-0.67) and bind to DNA by intercalation with high affinity (Kb = 1.5 × 106 M-1). Furthermore, under the same conditions, DNA-binding sempervirine derivatives were synthesized in a Westphal reaction from 1,2-diketones that have at least one non-enolizable α-carbon atom, which shows that the reaction pathway is determined by the substrate structure.

Studies on the Interactions of 3,11-Difluoro-6,8,13-trimethyl-8H-quino[4,3,2-kl]acridinium and Insulin with the Quadruplex-Forming Oligonucleotide Sequence a2 from the Insulin-Linked Polymorphic Region.

Recently, several quadruplex-DNA-forming sequences have been identified in the insulin-linked polymorphic region (ILPR), which is a guanine-rich oligonucleotide sequence in the promoter region of insulin. The formation of this non-canonical quadruplex DNA (G4-DNA) has been shown to be involved in the biological activity of the ILPR, specifically with regard to its interplay with insulin. In this context, this contribution reports on the investigation of the association of the quadruplex-forming ILPR sequence a2 with insulin as well as with the well-known G4-DNA ligand 3,11-difluoro-6,8,13-trimethyl-8H-quino[4,3,2-kl]acridinium (1), also named RHPS4, by optical and NMR spectroscopy. CD- and NMR-spectroscopic measurements confirmed the preferential formation of an antiparallel quadruplex structure of a2 with four stacked guanine quartets. Furthermore, ligand 1 has high affinity toward a2 and binds by terminal π stacking to the G1-G11-G15-G25 quartet. In addition, the spectroscopic studies pointed to an association of insulin to the deoxyribose backbone of the loops of a2.

DNA-Triggered Enhancement of Singlet Oxygen Production by Pyridinium Alkynylanthracenes.

There is an ongoing interest in 1 O2 sensitizers, whose activity is selectively controlled by their interaction with DNA. To this end, we synthesized three isomeric pyridinium alkynylanthracenes 2 o-p and a water-soluble trapping reagent for 1 O2 . In water and in the absence of DNA, these dyes show a poor efficiency to sensitize the photooxygenation of the trapping reagent as they decompose due to electron transfer processes. In contrast, in the presence of DNA 1 O2 is generated from the excited DNA-bound ligand. The interactions of 2 o-p with DNA were investigated by thermal DNA melting studies, UV/vis and fluorescence spectroscopy, and linear and circular dichroism spectroscopy. Our studies revealed an intercalative binding with an orientation of the long pyridyl-alkynyl axis parallel to the main axis of the DNA base pairs. In the presence of poly(dA : dT), all three isomers show an enhanced formation of singlet oxygen, as indicated by the reaction of the latter with the trapping reagent. With green light irradiation of isomer 2 o in poly(dA : dT), the conversion rate of the trapping reagent is enhanced by a factor >10. The formation of 1 O2 was confirmed by control experiments under anaerobic conditions, in deuterated solvents, or by addition of 1 O2 quenchers. When bound to poly(dG : dC), the opposite effect was observed only for isomers 2 o and 2 m, namely the trapping reagent reacted significantly slower. Overall, we showed that pyridinium alkynylanthracenes are very useful intercalators, that exhibit an enhanced photochemical 1 O2 generation in the DNA-bound state.

Synthesis of 10-O-aryl-substituted berberine derivatives by Chan-Evans-Lam coupling and investigation of their DNA-binding properties.

Eleven novel 10-O-aryl-substituted berberrubine and berberine derivatives were synthesized by the Cu2+-catalyzed Chan-Evans-Lam coupling of berberrubine with arylboronic acids and subsequent 9-O-methylation. The reaction is likely introduced by the Cu2+-induced demethylation of berberrubine and subsequent arylation of the resulting 10-oxyanion functionality. Thus, this synthetic route represents the first successful Cu-mediated coupling reaction of berberine substrates. The DNA-binding properties of the 10-O-arylberberine derivatives with duplex and quadruplex DNA were studied by thermal DNA denaturation experiments, spectrometric titrations as well as CD and LD spectroscopy. Fluorimetric DNA melting analysis with different types of quadruplex DNA revealed a moderate stabilization of the telomeric quadruplex-forming oligonucleotide sequence G3(TTAG3)3. The derivatives showed a moderate affinity towards quadruplex DNA (K b = 5-9 × 105 M-1) and ct DNA (K b = 3-5 × 104 M-1) and exhibited a fluorescence light-up effect upon complexation to both DNA forms, with slightly higher intensity in the presence of the quadruplex DNA. Furthermore, the CD- and LD-spectroscopic studies revealed that the title compounds intercalate into ct DNA and bind to G4-DNA by terminal stacking.

Berberrubine Phosphate: A Selective Fluorescent Probe for Quadruplex DNA.

A phosphate-substituted, zwitterionic berberine derivative was synthesized and its binding properties with duplex DNA and G4-DNA were studied using photometric, fluorimetric and polarimetric titrations and thermal DNA denaturation experiments. The ligand binds with high affinity toward both DNA forms (Kb = 2-7 × 105 M-1) and induces a slight stabilization of G4-DNA toward thermally induced unfolding, mostly pronounced for the telomeric quadruplex 22AG. The ligand likely binds by aggregation and intercalation with ct DNA and by terminal stacking with G4-DNA. Thus, this compound represents one of the rare examples of phosphate-substituted DNA binders. In an aqueous solution, the title compound has a very weak fluorescence intensity (Φfl < 0.01) that increases significantly upon binding to G4-DNA (Φfl = 0.01). In contrast, the association with duplex DNA was not accompanied by such a strong fluorescence light-up effect (Φfl < 0.01). These different fluorimetric responses upon binding to particular DNA forms are proposed to be caused by the different binding modes and may be used for the selective fluorimetric detection of G4-DNA.

Selective, pH-Dependent Colorimetric and Fluorimetric Detection of Quadruplex DNA with 4-Dimethylamino(phenyl)-Substituted Berberine Derivatives.

The 9- and 12-dimethylaminophenyl-substituted berberine derivatives 3 a and 3 b were readily synthesized by Suzuki-Miyaura reactions and shown to be useful fluorescent probes for the optical detection of quadruplex DNA (G4-DNA). Their association with the nucleic acids was investigated by spectrometric titrations, CD and LD spectroscopy, and with DNA-melting analysis. Both ligands bind to duplex DNA by intercalation and to G4-DNA by terminal π stacking. At neutral conditions, they bind with higher affinity (Kb =105 -106  M-1 ) to representative quadruplex forming oligonucleotides 22AG, c-myc, c-kit, and a2, than to duplex calf thymus (ct) DNA (Kb =5-7×104  M-1 ). At pH 5, however, the affinity of 3 a towards G4-DNA 22AG is higher (Kb =1.2×106  M-1 ), whereas the binding constant towards ct DNA is lower (Kb =3.9×103  M-1 ) than under neutral conditions. Notably, the association of the ligand with DNA results in characteristic changes of the absorption and emission properties under specific conditions, which may be used for optical DNA detection. Other than the parent berberine, the ligands do not show a noticeable increase of their very low intrinsic emission intensity upon association with DNA at neutral conditions. In contrast, a fluorescence light-up effect was observed upon association to duplex (Φfl =0.01) and quadruplex DNA (Φfl =0.04) at pH 5. This fluorimetric response to G4-DNA association in combination with the distinct, red-shifted absorption under these conditions provides a simple and conclusive optical detection of G4-DNA at lower pH.

Synthesis, DNA-binding and antiproliferative properties of diarylquinolizinium derivatives.

A series of ten 2,7- and 2,8-diarylquinolizinium derivatives was synthesized and their DNA-binding and cytotoxic properties were investigated. Except for one nitro-substituted derivative all tested diarylquinolizinium ions bind to DNA with sufficient affinity (2 × 104 M-1-2 × 105 M-1). It was shown with photometric, fluorimetric and polarimetric titrations as well as with flow-LD analysis that the ligands bind mainly by intercalation to duplex DNA, however, depending on the ligand-DNA ratio, groove binding and backbone association were also observed with some derivatives. The biological activity was further investigated with tests of cytotoxicity and antiproliferative properties towards non-tumor cells and selected cancer cells, along with cell cycle analysis and an annexin-V assay. Notably, substrates that carry donor-functionalities in the 4-position of the phenyl substituents revealed a strong, and in some cases selective, antiproliferative activity as quantified by the growth inhibition, GI50, at very low micromolar and even submicromolar level both in leukemia and solid tumors.

Fluorimetric Detection of Zn2+, Mg2+, and Fe2+ with 3-Hydroxy-4-Pyridylisoquinoline as Fluorescent Probe.

The suitability of 3-hydroxy-4-pyridylisoquinoline to operate as fluorescent chemosensor for the detection of metal ions was investigated. For that purpose, the interactions of the title compound with selected metal ions were investigated by absorption and emission spectroscopy. The complexation of Zn2+, Fe2+, Mg2+ with 1:1 and 2:1 stoichiometry leads to characteristic optical responses that depend significantly on the employed solvents, thus allowing for the fluorimetric identification and detection of particular metal cations in a matrix-based pattern analysis or by fluorimetric titrations.