Interaction of a bioactive pyrazole derivative with calf thymus DNA: deciphering the mode of binding by multi-spectroscopic and molecular docking investigations

Deoxyribonuclic acid (DNA) is the most relevant intracellular target for a wide variety of anticancer and antibiotic drugs. Elucidating the binding interaction of small bioactive molecules with DNA provides a structural guideline for designing new drugs with improved selectivity and superior clinical efficacy. In the present work interaction of a newly synthesized biologically relevant fluorophore, namely, (E)-1,5-diphenyl-3-styryl-4,5-dihydro-1H-pyrazole (DSDP) with calf thymus DNA (ctDNA) has been investigated vividly through a number of in vitro studies. Noteworthy modifications in the UV–Vis absorption and emission spectra reveal the formation of the probe–ctDNA complex. Several other spectroscopic experiments such as circular dichromism (CD), iodide induced quenching, competitive binding assay with known groove binder probe, 3-hydroxyflavone (3HF), time resolved fluorescence decay measurements, thermometric experiment in connection with the helix melting of ctDNA etc. unequivocally ascertain the groove binding interaction of DSDP with ctDNA. Determination of the thermodynamic parameters through temperature variation study implies the dominant role of hydrophobic interaction in the probe―DNA binding process. Inappreciable change in the CD spectrum of ctDNA with the addition of DSDP suggests that binding of the probe with the DNA does not lead to a significant modification in the DNA conformation. In-silico molecular docking simulation corroborates the experimental findings and depicts that DSDP favorably binds to the minor groove region of the biomacromolecule

Photophysics of α-furil at room temperature and 77 K: spectroscopic and quantum chemical studies

Steady state and time resolved spectroscopic measurements have been exploited to assign the emissions from different conformations of α-furil (2, 2′-furil) in solution phase at room temperature as well as cryogen (liquid nitrogen, LN2) frozen matrices of ethanol and methylcyclohexane. Room temperature studies reveal a single fluorescence from the trans-planar conformer of the fluorophore or two fluorescence bands coming from the trans-planar and the relaxed skew forms depending on excitation at the nπ* or the ππ* absorption band, respectively. Together with the fluorescence bands, the LN2 studies in both the solvents unambiguously ascertain two phosphorescence emissions with lifetimes 5 ± 0.3 ms (trans-planar triplet) and 81 ± 3 ms (relaxed skew triplet). Quantum chemical calculations have been performed using density functional theory at CAM-B3LYP/6-311++G** level to prop up the spectroscopic surveillance. The simulated potential energy curves (PECs) illustrate that α-furil is capable of giving two emissions from each of the S1 and the T1 states—one corresponding to the trans-planar and the other to the relaxed skew conformation. Contrary to the other 1,2-dicarbonyl molecular systems like benzil and α-naphthil, α-furil does not exhibit any fluorescence from its second excited singlet (S2) state. This is ascribed to the proximity of the minimum of the PEC of the S2 state and the hill-top of the PEC of the S1 state

Exploration of the binding interaction of a potential nervous system stimulant with calf-thymus DNA and dissociation of the drug–DNA complex by detergent sequestration

The binding interaction of a potential nervous system stimulant, 3-acetyl-4-oxo-6,7-dihydro-12H-indolo-[2,3-a]-quinolizine (AODIQ), with calf-thymus DNA (ctDNA) has been explored thoroughly. The modified photophysics of the fluorescent molecule within the microheterogeneous biomacromolecular system has been exploited to divulge the drug–DNA binding interaction. The absorption and various fluorometric measurements together with the fluorescence quenching, urea-induced denaturation study, circular dichroism and DNA-melting studies unequivocally ascertain the mode of binding of the drug with the DNA to be groove binding. Corroborating the experimental observations, molecular docking simulation projects the minor groove of the biomacromolecule to be the site of binding. Further experiments have revealed that dissociation of the drug from the drug–DNA complex can be achieved by the detergent sequestration method. Besides providing an insight into the drug–DNA interaction the work also demonstrates the surfactant-induced excretion of the drug from the biomacromolecular assembly

Exploration of photophysics of 2,2′-pyridil at room temperature and 77 K: a combined spectroscopic and quantum chemical approach

The photophysics of 2,2′-pyridil has been explored thoroughly using steady state and time resolved fluorometric techniques at room temperature (RT) in liquid media as well as in glassy matrices at cryogenic temperature (77 K). Ethanol and methylcyclohexane are exploited for this purpose, as polar and non-polar media respectively. Notwithstanding the observation of multiple emissions from the fluorophore, the experiments unequivocally rule out emission from excited singlet states other than the S1 state, consistent with Kasha's rule. Among 1,2-dicarbonyl molecular systems, this behavior resembles that of α-furil, while it contradicts that of benzil and α-naphthil which exhibit S2 emissions. The dual fluorescence and dual phosphorescence of the fluorophore are ascribed to the emissions originating from the two conformers, namely near-trans and relaxed skew. Coexistence of the two conformers is substantiated by time resolved area normalized emission spectroscopy (TRANES) at both RT and 77 K. The potential energy curves (PECs) simulated from calculations based on density functional theory and its time dependent extension provide adequate support to the experimental observations

Cyclodextrin induced controlled delivery of a biological photosensitizer from a nanocarrier to DNA

In this article, we have addressed to a demanding physicochemical aspect of therapeutic and drug research. We have reported a simple yet prospective technique that can be exploited for the controlled delivery of drugs and/or bioactive small molecules to the most relevant biomolecular target DNA. Exploiting various steady state and time resolved spectroscopic techniques together with the DNA helix melting study, we have shown that a biologically significant photosensitizer, namely, phenosafranin (PSF), can be quantitatively transferred to the DNA from the micellar nanocarrier made up of sodium tetradecyl sulfate (STS) using the external stimulant β-cyclodextrin (β-CD). The complexation property of β-CD with the nanocarrier (STS) has been utilized for the controlled release of the probe from the micelle to the DNA. Non-toxicity of the stimulant and the noninvasive nature of the carrier towards the target are expected to add to the suitability of this approach from a clinical perspective

DNA induced sequestration of a bioactive cationic fluorophore from the lipid environment: a spectroscopic investigation

The effect of calf-thymus DNA (ctDNA) on the lipid bound probe, formed by the cationic phenazinium dye phenosafranin (PSF) and the anionic lipid dimyristoyl-L-α-phosphatidylglycerol (DMPG), has been unearthed exploiting various spectroscopic techniques. Steady state and time-resolved fluorometric studies and measurements of circular dichroism and DNA helix melting temperature reveal that in the presence of DNA the probe is dislodged from the lipid environment and gets intercalated within the DNA helix. The work qualitatively illustrates that the anionic lipid can be used as a potential nanocarrier for delivering the cationic drugs to the most relevant biomacromolecular target, DNA

Delivery of a bioactive photosensitizer to natural DNA using γ-cyclodextrin as carrier

147-156A challenging goal of targeted drug delivery is to discover novel administrating route for the delivery of bioactive molecules/drugs in the field of pharmaceutical, medicinal and biophysical research. Delivery of a cationic photosensitizer, namely, phenosafranin (PSF), to the most relevant biomolecular target DNA using nanocarrier, γ-cyclodextrin (γ-CD), has been explored via various steady state and time resolved fluorometric as well as other spectroscopic techniques. The detailed fluorometric studies including DNA helix melting experiments divulge that the binding affinity of probe with the target (DNA) is order of magnitude larger than with the carrier (γ-CD). This leads to the release of the probe from the carrier cyclodextrin to bind with the DNA. Endogenous activation, in terms of competitive binding affinity, has been exploited for this purpose. This work qualitatively illustrates that the γ-cyclodextrin can be used as a safe and potential drug delivery system (DDS) for drug targeting towards DNA

Binding of an anionic fluorescent probe with calf thymus DNA and effect of salt on the probe–DNA binding: a spectroscopic and molecular docking investigation

Binding interaction of a biologically relevant anionic probe molecule, 8-anilino-1-naphthalene sulfonate (ANS) with calf-thymus deoxyribonucleic acid (ctDNA) has been investigated exploiting vivid spectroscopic techniques together with molecular docking study. Significant modifications in the absorption and emission profiles, the determined binding constant, micropolarity analysis, circular dichroism (CD) spectral study, comparative binding study with ethidium bromide (EtBr)—an intercalative binder, thermometric experiment relating to the helix melting of ctDNA and blind molecular docking simulation confirm the groove binding of ANS with ctDNA. Furthermore, a remarkable enhancement is observed in the fluorescence intensity as well as in the fluorescence lifetime of the DNA-bound probe with the addition of salts. Reduction in the electrostatic repulsion between the ANS and DNA at high salt concentration has been assigned responsible for this observation. Besides providing an insight into the probe–DNA interaction, the work implies that the binding interaction of a negatively charged probe with DNA can be enhanced considerably by the addition of salts

Binding interaction of differently charged fluorescent probes with egg yolk phosphatidylcholine and the effect of β-cyclodextrin on the lipid–probe complexes: a fluorometric investigation

Interaction of cationic phenosafranin (PSF), anionic 8-anilino-1-naphthalene sulfonate (ANS) and non-ionic nile red (NR) have been studied with the zwitterionic phospholipid, egg yolk L-α-phosphatidylcholine (EYPC). The study reveals discernible binding interactions of the three fluorescent probes with the EYPC lipid vesicle. Once the binding of the probes with the lipid is established, the effect of cyclic oligosaccharide, β-cyclodextrin (β-CD), on these lipid bound probes has been investigated. Different fluorometric techniques suggest that addition of β-CD to the probe–lipid complexes leads to the release of the probes from the lipid medium through the formation of probe–β-CD inclusion complexes. A competitive binding of the probes between β-cyclodextrin and the lipid is ascribed to be responsible for the effect. This provides an easy avenue for the removal of the probe molecules from the lipid environment. Extension of this work with drug molecules in cell membranes is expected to give rise to a strategy for the removal of adsorbed drugs from the cell membranes by the use of non-toxic β-cyclodextrin

Impact of structural modification on the photophysical response of benzoquinoline fluorophores

Structural influence on the photophysical behavior of two pairs of molecular systems from the biologically potent benzoquinoline family, namely, dimethyl-3-(4-chlorophenyl)-3,4-dihydrobenzo[f]-quinoline-1,2-dicarboxylate, dimethyl-3-(2,6-dichlorophenyl)-3,4-dihydrobenzo[f]quinoline-1,2-dicarboxylate and their corresponding dehydrogenated analogues has been investigated exploiting experimental as well as computational techniques. The study unveils that dehydrogenation in the heterocyclic rings of the studied quinoline derivatives modifies their photophysics radically. Experimental observations imply that the photophysical behavior of the dihydro analogues is governed by the intramolecular charge transfer (ICT) process. However, the ICT process is restricted significantly by the dehydrogenation of the heterocyclic rings. Computational exertion leads to the proposition that the change in the electronic distribution in these molecular systems on dehydrogenation is the rationale behind the dramatic modification of their photophysics