P-stereocontrolled synthesis of oligo(nucleoside N3′→O5′ phosphoramidothioate)s – opportunities and limitations

3′-N-(2-Thio-1,3,2-oxathiaphospholane) derivatives of 5′-O-DMT-3′-amino-2′,3′-dideoxy-ribonucleosides (_NOTP-N), that bear a 4,4-unsubstituted, 4,4-dimethyl, or 4,4-pentamethylene substituted oxathiaphospholane ring, were synthesized. Within these three series, _NOTP-N differed by canonical nucleobases (i.e., Ade^Bz, Cyt^Bz, Gua^iBu, or Thy). The monomers were chromatographically separated into P-diastereomers, which were further used to prepare N_NPSN′ dinucleotides (3), as well as short P-stereodefined oligo(deoxyribonucleoside N3′→O5′ phosphoramidothioate)s (NPS-) and chimeric NPS/PO- and NPS/PS-oligomers. The condensation reaction for _NOTP-N monomers was found to be 5–6 times slower than the analogous OTP derivatives. When the 5′-end nucleoside of a growing oligomer adopts a C3′-endo conformation, a conformational ‘clash’ with the incoming _NOTP-N monomer takes place, which is a main factor decreasing the repetitive yield of chain elongation. Although both isomers of N_NPSN′ were digested by the HINT1 phosphoramidase enzyme, the isomers hydrolyzed at a faster rate were tentatively assigned the R_P absolute configuration. This assignment is supported by X-ray analysis of the protected dinucleotide ^DMTdG^iBu_NPSMeT_OAc, which is P-stereoequivalent to the hydrolyzed faster P-diastereomer of dG_NPST.

Separated P-diastereomers of 3′-N-(2-thio-1,3,2-oxathiaphospholane) derivatives of 5′-O-DMT-3′-amino-2′,3′-dideoxy-ribonucleosides were used to prepare P-stereodefined N_NPSN′ dinucleotides and short NPS-, NPS/PO- and NPS/PS-oligomers.     

[2,2′-Bithiophene]-4,4′-dicarboxamide: a novel building block for semiconducting polymers

A novel electron deficient building block [2,2′-bithiophene]-4,4′-dicarboxamide (BTDCA) was designed to lower the highest occupied molecular orbital (HOMO) energy level of polythiophenes in order to achieve a higher open circuit voltage (V_oc) and thus a higher power conversion efficiency in polymer solar cells (PSCs). BTDCA dibromo monomers were conveniently synthesized in four steps, and were used to prepare three thiophene-based D-A polymers, P(BTDCA66-BT) (66BT), P(BTDCA44-BT) (44BT) and P(BTDCA44-TT) (44TT). All the polymers exhibited unipolar hole transport properties, exhibiting mobilities in the range of ∼10⁻⁴ to 10⁻² cm² V⁻¹ s⁻¹ with the highest hole mobility of up to 1.43 × 10⁻² cm² V⁻¹ s⁻¹ achieved for 44BT in bottom-gate bottom-contact organic thin film transistors (OTFTs). In PSCs, these polymers achieved high V_oc''s of 0.81–0.87 V when PCBM or ITIC was used as acceptor. When 44TT was used as donor and ITIC was used as acceptor, a power conversion efficiency (PCE) of up to 4.5% was obtained, a significant improvement when compared with the poly(3-hexylthiophene) (P3HT):ITIC devices, which showed the highest PCE of merely 0.92%.

A new electron acceptor building block, [2,2′-bithiophene]-4,4′-dicarboxamide, is synthesized and used to develop donor polymers for organic solar cells.     

Antiherpesvirus Activities of (1′S,2′R)-9-{[1′,2′-Bis(hydroxymethyl)cycloprop-1′-yl]methyl}guanine (A-5021) in Cell Culture

Antiherpetic activity of (1′S,2′R)-9-{[1′,2′-bis(hydroxymethyl)cycloprop-1′-yl]methyl}guanine (A-5021) was compared with those of acyclovir (ACV) and penciclovir (PCV) in cell cultures. In a plaque reduction assay using a selection of human cells, A-5021 showed the most potent activity in all cells. Against clinical isolates of herpes simplex virus type 1 (HSV-1, n = 5) and type 2 (HSV-2, n = 6), mean 50% inhibitory concentrations (IC₅₀s) for A-5021 were 0.013 and 0.15 μg/ml, respectively, in MRC-5 cells. Corresponding IC₅₀s for ACV were 0.22 and 0.30 μg/ml, and those for PCV were 0.84 and 1.5 μg/ml, respectively. Against clinical isolates of varicella-zoster virus (VZV, n = 5), mean IC₅₀s for A-5021, ACV, and PCV were 0.77, 5.2, and 14 μg/ml, respectively, in human embryonic lung (HEL) cells. A-5021 showed considerably more prolonged antiviral activity than ACV when infected cells were treated for a short time. The selectivity index, the ratio of 50% cytotoxic concentration to IC₅₀, of A-5021 was superior to those of ACV and PCV for HSV-1 and almost comparable for HSV-2 and VZV. In a growth inhibition assay of murine granulocyte-macrophage progenitor cells, A-5021 showed the least inhibitory effect of the three compounds. These results show that A-5021 is a potent and selective antiviral agent against HSV-1, HSV-2, and VZV.     

Enzymatic Adenylylation by Aminoglycoside 4′-Adenylyltransferase and 2′′-Adenylyltransferase as a Means of Determining Concentrations of Aminoglycoside Antibiotics in Serum

A fast and accurate enzymatic estimation of the concentration of aminoglycoside antiobiotics in serum is given by aminoglycoside 4′-adenylyltransferase (synonym: tobramycin adenylyltransferase), a new enzyme recently found in tobramycin-resistant staphylococci (P. Santanam and F. H. Kayser, J. Infect. Dis. [Suppl.], in press). This enzyme is useful in assaying the kanamycins, the butirosins, amikacin, ribostamycin and, if required, the neomycins, tobramycin, and paromomycin. In combination with the assay using aminoglycoside 2′′-adenylyltransferase (synonym: gentamicin adenylyltransferase) to determine the level of gentamicin, sisomicin, and tobramycin, a convenient method to estimate a wide range of aminoglycoside antibiotics in patients' sera is described.     

Theoretical study of D–A′–π–A/D–π–A′–π–A triphenylamine and quinoline derivatives as sensitizers for dye-sensitized solar cells

We have designed four dyes based on D–A′–π–A/D–π–A′–π–A triphenylamine and quinoline derivatives for dye-sensitized solar cells (DSSCs) and studied their optoelectronic properties as well as the effects of the introduction of alkoxy groups and thiophene group on these properties. The geometries, single point energy, charge population, electrostatic potential (ESP) distribution, dipole moments, frontier molecular orbitals (FMOs) and HOMO–LUMO energy gaps of the dyes were discussed to study the electronic properties of dyes based on density functional theory (DFT). And the absorption spectra, light harvesting efficiency (LHE), hole–electron distribution, charge transfer amount from HOMO to LUMO (Q_CT), D index, H_CT index, S_m index and exciton binding energy (E_coul) were discussed to investigate the optical and charge-transfer properties of dyes by time-dependent density functional theory (TD-DFT). The calculated results show that all the dyes follow the energy level matching principle and have broadened absorption bands at visible region. Besides, the introduction of alkoxy groups into triarylamine donors and thiophene groups into conjugated bridges can obviously improve the stability and optoelectronic properties of dyes. It is shown that the dye D4, which has had alkoxy groups as well as thiophene groups introduced and possesses a D–π–A′–π–A configuration, has the optimal optoelectronic properties and can be used as an ideal dye sensitizer.

We have designed four dyes based on D–A′–π–A/D–π–A′–π–A triphenylamine and quinoline derivatives for DSSCs and studied their optoelectronic properties as well as the effects of the introduction of alkoxy groups and thiophene group on the properties.     

Molecular dynamics simulations of a cyclotetramethylene tetra-nitramine/hydrazine 5,5′-bitetrazole-1,1′-diolate cocrystal

An energetic ionic salt (EIS)-based cocrystal formation, cyclotetramethylene tetra-nitramine (HMX)/hydrazine 5,5′-bitetrazole-1,1′-diolate (HA·BTO), is predicted based on molecular dynamics simulations. HA·BTO is a newly-synthesized environmentally friendly energetic ionic salt with good detonation performance and low sensitivity. Calculated powder X-ray diffraction patterns and intermolecular interactions deduce the formation of the new cocrystal structure. Radial distribution function analysis suggests that hydrogen bonds and van der Waals (vdW) forces exist between the H⋯O pairs of HMX and HA·BTO, while the hydrogen bonds between the H of HA·BTO and the O of HMX play a prominent role. The cohesive energy density and mechanical properties are also analyzed. The cohesive energy density of the HMX/HA·BTO cocrystal is larger than that of the composite of HMX and HA·BTO, indicating an improvement in crystal stability by cocrystalization. Compared to both HMX and HA·BTO, HMX/HA·BTO has smaller Young modulus, bulk modulus and shear modulus values, but larger K/G values and a positive Cauchy pressure, suggesting decreased stiffness and improved ductibility. Moreover, the calculated formation energy is −405.79 kJ mol⁻¹ at 298 K, which implies that the proposed cocrystal structure is likely to be synthesized at ambient temperature. In summary, we have predicted an EIS-based cocrystal formation in which the safety and mechanical properties of HMX have been improved via cocrystalization with HA·BTO, and this provides deep insight into the formation mechanism of the EIS-based cocrystal.

An energetic ionic salt-based cocrystal formation, HMX/HA·BTO, is predicted based on molecular dynamics simulations.     

Reactive molecular dynamics simulation of the high-temperature pyrolysis of 2,2′,2′′,4,4′,4′′,6,6′,6′′-nonanitro-1,1′:3′,1′′-terphenyl (NONA)

2,2′,2′′,4,4′,4′′,6,6′,6′′-Nonanitro-1,1′:3′,1′′-terphenyl (NONA) is currently recognized as an excellent heat-resistant explosive. To improve the atomistic understanding of the thermal decomposition paths of NONA, we performed a series of reactive force field (ReaxFF) molecular dynamics simulations under extreme conditions of temperature and pressure. The results show that two distinct initial decomposition mechanisms are the homolytic cleavage of the C–NO₂ bond and nitro–nitrite (NO₂ → ONO) isomerization followed by NO fission. Bimolecular and fused ring compounds are found in the subsequent decomposition of NONA. The product identification analysis under finite time steps showed that the gaseous products are CO₂, N₂, and H₂O. The amount of CO₂ is energetically more favorable for the system at high temperature or low density. The carbon-containing clusters are a favorable growth pathway at low temperatures, and this process was further demonstrated by the analysis of diffusion coefficients. The increase of the crystal density accelerates the decomposition of NONA judged by the analysis of reaction kinetic parameters and activation barriers. In the endothermic and exothermic stages, a 20% increase in NONA density increases the activation energies by 3.24 and 0.48 kcal mol⁻¹, respectively. The values of activation energies (49.34–49.82 kcal mol⁻¹) agree with the experimental data in the initial decomposition stage.

The bimolecular and fused ring compounds are found in the high-temperature pyrolysis of NONA using ReaxFF molecular dynamics simulations.     

A promising TNT alternative 3,3′-bi(1,2,4-oxadiazole)-5,5′-diylbis(methylene)dinitrate (BOM): thermal behaviors and eutectic characteristics

3,3′-Bi(1,2,4-oxadiazole)-5,5′-diylbis(methylene)dinitrate (BOM) is a liquid phase carrier for melt cast explosives that is expected to replace TNT. The combination of a conjugated 1,2,4-oxadiazole backbone and nitrate ester groups endows BOM with both good energetic performance and impressive insensitivity. In this paper, the thermal behaviors of BOM were investigated using a TG–DSC synchronous thermal analyzer, proving that BOM is basically non-volatile under heating and melting processes. The apparent activation energy of BOM calculated by the Kissinger method was 158.2 kJ mol⁻¹ at atmospheric pressure, which is higher than that of DNTF at atmospheric pressure and TNT at 2 MPa, indicating good thermal stability at low temperatures. The thermal decomposition mechanism of BOM was studied through both DSC-MS and in situ FTIR technologies. The low eutectic characteristics of BOM and DNTF were also investigated carefully and the best ratio of BOM/DNTF was 40/60 with a melting point at 75.5 °C. Finally, the detonation performances of TNT/HMX, BOM/HMX and BOM/DNTF(40/60)/HMX explosive formulations were calculated, showing that the detonation performances of the latter two formulations were significantly higher than that of TNT/HMX.

3,3′-Bi(1,2,4-oxadiazole)-5,5′-diylbis(methylene)dinitrate (BOM) is a liquid phase carrier for melt cast explosives that is expected to replace TNT.     

The modulation effect of pi–pi interactions on the electronic and photochromic properties of viologen complexes containing N,N′-bis(carboxyethyl)-4,4′-bipyridinium

Two viologen complexes containing N,N′-bis(carboxyethyl)-4,4′-bipyridinium (BCEbpy) were prepared, namely [Zn(H₂O)₆]·(BCEbpy)·(p-BDC)·3H₂O (1) and [Co(H₂O)₆]·(BCEbpy)·(p-BDC)·3H₂O (2) (p-H₂BDC = 1,4-benzenedicarboxylic acid), and their crystal structures, photochromism, frontier molecular orbitals, Hirshfeld surfaces and 2D fingerprint plots were investigated. The modulation effects of pi–pi interactions were explored on the electronic and photochromic properties of compounds 1 and 2. Due to the existence of photo-response viologen radicals, both complexes 1 and 2 display excellent photo-response properties in the sequence 1 < 2. The results indicate that compound 1 exhibits intramolecular electron transfer; compound 2 exhibits both intramolecular and intermolecular electron transfer, which is mainly due to the change of electronic and steric structures caused by pi–pi interactions with a faster photo-response rate than that of compound 1. The donor–acceptor modes, matching principles and inter/intramolecular atom–atom close contacts were illustrated by the density functional theory (DFT)-B3LYP/6-311(d,p) method.

Two viologen complexes containing BCEbpy were prepared and displayed excellent photo-response properties by the modulation effect of pi–pi interactions.     

Mode of Action of (1′S,2′R)-9-{[1′,2′-Bis(hydroxymethyl) cycloprop-1′-yl]methyl}guanine (A-5021) against Herpes Simplex Virus Type 1 and Type 2 and Varicella-Zoster Virus

The mode of action of (1′S,2′R)-9-{[1′,2′-bis(hydroxymethyl)cycloprop-1′-yl]methyl}guanine (A-5021) against herpes simplex virus type 1 (HSV-1), HSV-2, and varicella-zoster virus (VZV) was studied. A-5021 was monophosphorylated at the 2′ site by viral thymidine kinases (TKs). The 50% inhibitory values for thymidine phosphorylation of A-5021 by HSV-1 TK and HSV-2 TK were comparable to those for penciclovir (PCV) and lower than those for acyclovir (ACV). Of these three agents, A-5021 inhibited VZV TK most efficiently. A-5021 was phosphorylated to a mono-, di-, and triphosphate in MRC-5 cells infected with HSV-1, HSV-2, and VZV. A-5021 triphosphate accumulated more than ACV triphosphate but less than PCV triphosphate in MRC-5 cells infected with HSV-1 or VZV, whereas HSV-2-infected MRC-5 cells had comparable levels of A-5021 and ACV triphosphates. The intracellular half-life of A-5021 triphosphate was considerably longer than that of ACV triphosphate and shorter than that of PCV triphosphate. A-5021 triphosphate competitively inhibited HSV DNA polymerases with respect to dGTP. Inhibition was strongest with ACV triphosphate, followed by A-5021 triphosphate and then (R,S)-PCV triphosphate. A DNA chain elongation experiment revealed that A-5021 triphosphate was incorporated into DNA instead of dGTP and terminated elongation, although limited chain extension was observed. Thus, the strong antiviral activity of A-5021 appears to depend on a more rapid and stable accumulation of its triphosphate in infected cells than that of ACV and on stronger inhibition of viral DNA polymerase by its triphosphate than that of PCV.     

Organic binary charge-transfer compounds of 2,2′ : 6′,2′′ : 6′′,6-trioxotriphenylamine and a pyrene-annulated azaacene as donors

Three binary charge-transfer (CT) compounds resulting from the donor 2,2′ : 6′,2′′ : 6′′,6-trioxotriphenylamine (TOTA) and the acceptors F₄TCNQ and F₄BQ and of a pyrene-annulated azaacene (PAA) with the acceptor F₄TCNQ are reported. The identity of these CT compounds are confirmed by single-crystal X-ray diffraction as well as by IR, UV-vis-NIR and EPR spectroscopy. X-ray diffraction analysis reveals a 1 : 1 stoichiometry for TOTA·F₄TCNQ, a 2 : 1 donor : acceptor ratio in (TOTA)₂·F₄BQ, and a rare 4 : 1 stoichiometry in (PAA)₄·F₄TCNQ, respectively. Metrical parameters of the donor (D) and acceptor (A) constituents as well as IR spectra indicate full CT in TOTA·F₄TCNQ, partial CT in (TOTA)₂·F₄BQ and only a very modest one in (PAA)₄·F₄TCNQ. Intricate packing motifs are present in the crystal lattice with encaged, π-stacked (F₄TCNQ⁻)₂ dimers in TOTA·F₄TCNQ or mixed D/A stacks in the other two compounds. Their solid-state UV-vis-NIR spectra feature CT transitions. The CT compounds with F₄TCNQ are electrical insulators, while (TOTA)₂·F₄BQ is weakly conducting.

Three binary charge-transfer (CT) compounds resulting from the donor 2,2′ : 6′,2′′ : 6′′,6-trioxotriphenylamine (TOTA) and the acceptors F₄TCNQ and F₄BQ and of a pyrene-annulated azaacene (PAA) with the acceptor F₄TCNQ are reported.     

Poly[2,2′-(4,4′-bipyridine)-5,5′-bibenzimidazole] functionalization of carbon black for improving the oxidation stability and oxygen reduction reaction of fuel cells

The rapid oxidation of carbon black (CB) is a major drawback for its use as a catalyst support in polymer electrolyte fuel cells. Here, we synthesize poly[2,2′-(4,4′-bipyridine)-5,5′-bibenzimidazole] (BiPyPBI) as a conducting polymer and use it to functionalize the surface of CB and homogenously anchor platinum metal nanoparticles (Pt-NPs) on a CB surface. The as-prepared materials were confirmed by different spectroscopic techniques, including nuclear magnetic resonance spectroscopy, energy-dispersive X-ray, thermal gravimetric analysis, and scanning-transmittance microscopy. The as-fabricated polymer-based CB catalyst showed an electrochemical surface area (ECSA) of 63.1 cm² mg_Pt⁻¹, giving a catalyst utilization efficiency of 74.3%. Notably, the BiPyPBI-based CB catalyst exhibited remarkable catalytic activity towards oxygen reduction reactions. The onset potential and the diffusion-limiting current density reached 0.66 V and 5.35 mA cm⁻², respectively. Furthermore, oxidation stability testing showed a loss of only 16% of Pt-ECSA for BiPyPBI-based CB compared to a 36% loss of Pt-ECSA for commercial Pt/CB after 5000 potential cycles. These improvements were related to the synergetic effect between the nitrogen-rich BiPyPBI polymer, which promoted the catalytic activity through the structural nitrogen atoms, and demolished the degradation of CB via the wrapping process.

The rapid oxidation of carbon black (CB) is a major drawback for its use as a catalyst support in polymer electrolyte fuel cells.     

2′-Deoxy-5-(hydroxymethyl)cytidine: estimation in human cancer cells with a simple chemosensor

A pyrrole-based rhodamine conjugate (CS-1) has been developed and characterized for the selective detection and quantification of 2′-deoxy-5-(hydroxymethyl)cytidine (5hmC) in human cancer cells with a simple chemosensing method.

A new chemosensor, CS-1, has been developed and characterized for the selective detection and quantification of 2′-deoxy-5-(hydroxymethyl)cytidine (5hmC) in human cancer cells.

2′-Deoxy-5-(hydroxymethyl)cytidine (5hmC) is found in both neuronal cells and embryonic stem cells. It is a modified pyrimidine and used to quantify DNA hydroxymethylation levels in biological samples^1–3 as it is capable of producing interstrand cross-links in double-stranded DNA. It is produced through an enzymatic pathway carried out by the Ten-Eleven Translocation (TET1, TET2, TET3) enzymes, iron and 2-oxoglutarate dependent dioxygenase.^4–7 In the DNA demethylation process, methylcytosine is converted to cytosine and generates 5hmC as an intermediate in the first step of this process which is then further oxidized to 5-formylcytosine (fC) and 5-carboxycytosine (caC) of very low levels compared to the cytosine level.⁸ Though the biological function of 5hmC in the mammalian genome is still not revealed, the presence of a hydroxymethyl group can regulate gene expression (switch ON & OFF). Reports say that in artificial DNA 5hmC is converted to unmodified cytosine when introduced into mammalian cells.^9,10Levels of 5hmC substantially vary in different tissues and cells. It is found to be highest in the brain, particularly in nervous system and in moderate percentage in liver, colon, rectum and kidney tissues, whereas it is relatively low in lung and very low in breast and placenta.^11,12 The percentage of 5hmC content is much less in cancer and tumor tissues compared to the healthy ones. The reason behind this loss is the absence of TET1, TET2, TET3, IDH1, or IDH2 mutations in most of the human cancer cells which means decrease of methylcytosine oxidation.^13–15 This loss of 5hmC in cancer cells is being used as a diagnostic tool for the detection of early-stage of malignant disease. Few analytical methods^16–19 such as glucosyltransferase assays, tungsten-based oxidation systems, and TET-assisted bisulfite sequencing (TAB-Seq) or oxidative bisulfite sequencing (oxBS-Seq) protocols are now developed to differentiate 5hmC from other nucleotide which are naturally occurred. There are also few methods such as liquid chromatography/tandem mass spectroscopy (LC/MS-MS), which determine the level of 5hmC in mammalian cancer cell.^20–22 However, these procedures are highly toxic and expensive due to requirement of catalyzation through enzymes or heavy metal ion and these techniques require expertise, facilities, much time and costs even beyond standard DNA sequencing. As a result, these detection techniques are currently inappropriate for the high-throughput screening of genome-wide 5hmC levels (performance comparison is shown in Table S1, ESI^†).Among all reputed methods fluorescence detection method using chemosensors is significantly important due to its indispensable role in medicinal and biological applications.^23–27 Chemosensors have been effectively explored to monitor biochemical processes and assays through in situ analysis in living systems and abiotic samples with much less time and cost.In this contribution we prepared and characterize (Scheme S1 and Fig. S1–S3, ESI^†) a pyrrole–rhodamine based chemosensor (CS-1) which shows efficient and selective fluorescence signal for 5hmC in aqueous medium (Scheme 1). A transparent single crystal of CS-1 (Fig. 1) was obtained by slow evaporation of the solvent from a solution of CS-1 in CH₃CN. It crystallizes as monoclinic with space group P2₁/n (Fig. S4 and Table S2, ESI^†).Open in a separate windowScheme 15hmC-induced FRET OFF–ON mechanism of the chemosensor CS-1.Open in a separate windowFig. 1ORTEP diagram of CS-1 (ellipsoids are drawn at 40% probability level).Spectrophotometric and spectrofluorimetric titrations were carried out to understand the CS-1–5hmC interaction with 1 : 1 binding stoichiometry (Fig. S5, ESI^†) upon adding varying concentrations of 5hmC to a fixed concentration of CS-1 (1 μM) in aqueous medium at neutral pH. Upon the addition of increasing concentrations of the 5hmC, a clear absorption band (K_a = 4.47 × 10⁵ M⁻¹, Fig. S6, ESI^†) appeared to be centered at 556 nm with increasing intensity (Fig. 2a). On the other hand, for the fluorescence emission spectra of CS-1 (Fig. 2b), upon irradiation at 325 nm, an emission maxima at 390 nm was observed, which was attributed to the fluorescence emission from the donor unit i.e. the pyrrole moiety of CS-1. When 5hmC were added, due to rhodamine moiety CS-1 showed a 95-fold increase in fluorescence at 565 nm (K_a = 4.61 × 10⁵ M⁻¹, Fig. S7, ESI^†) with the detection limit of 8 nM (Fig. S8, ESI^†). The binding of 5hmC induces opening of the spirolactam ring in CS-1, inducing a shift of the emission spectrum. Subsequently, increased overlap between the emission of the energy-donor (pyrrole) and the absorption of the energy-acceptor (rhodamine) greatly enhances the intramolecular FRET process,^28,29 producing an emission from the energy acceptor unit in CS-1.Open in a separate windowFig. 2(a) UV-vis absorption spectra of CS-1 (1 μM) upon gradual addition of 5hmC up to 1.2 equiv. in H₂O–CH₃CN (15 : 1, v/v) at neutral pH. (b) Fluorescence emission spectra of CS-1 (1 μM) upon addition of 1.2 equiv. of 5hmC in H₂O–CH₃CN (15 : 1, v/v) at neutral pH (λ_ex = 325 nm).In order to establish the sensing selectivity of the chemosensor CS-1, parallel experimentations were carried out with other pyrimidine/purine derivatives such as 5-methylcytosine, cytosine, cytidine, thymine, uracil, 5-hydroxymethyluracil, adenine and guanine. Comparing with other pyrimidine/purine derivatives the abrupt fluorescence enhancement was found upon addition of 5hmC to CS-1 while others do not make any fluorescence changes under UV lamp (Fig. 3, lower panel). Furthermore, the prominent color change from colorless to deep pink allows 5hmC to be detected by naked eye (Fig. 3, upper panel). The above observation shows consistency with the fluorescence titration experiments where no such binding of CS-1 with other pyrimidine/purine derivatives was found (Fig. S9, ESI^†).Open in a separate windowFig. 3Visible color (top) and fluorescence changes (bottom) of CS-1 (1 μM) in aqueous medium upon addition of 1.2 equiv. of various pyrimidine/purine derivatives (λ_ex = 325 nm) in H₂O–CH₃CN (15 : 1, v/v) at neutral pH.pH titration reveals that CS-1 becomes fluorescent below pH 5 due to the spirolactam ring opening of rhodamine. However, it is non-fluorescent at pH range of 5–13. Upon addition of 5hmC to CS-1 shows deep red fluorescence in the pH range of 5–8 (Fig. S10, ESI^†). Considering the biological application and the practical applicability of the chemosensor pH 7.4 has been preferred to accomplish all experiments successfully.In ¹H NMR titration (Fig. S11, ESI^†), the most interesting feature is the continuous downfield shift of aromatic protons on the pyrrole moiety of CS-1 upon gradual addition of 5hmC. This may be explained as the decrease in electron density of the pyrrole moiety upon binding with 5hmC through hydrogen bonding. Xanthene protons to be shifted downfield upon spirolactam ring opening indicates the probe to coordinate with 5hmC and electrons are accumulated around 5hmC. In ¹³C NMR titration the spiro cycle carbon peak at 65 ppm was shifted to 138 ppm along with a little downfield shift of the aromatic region of CS-1 (Fig. S12, ESI^†). This coordination led to the spiro cycle opening and changes to the absorption and emission spectra, further evident by mass spectrometry (Fig. S13, ESI^†), which corroborates the stronger interaction of CS-1 with 5hmC.The experimental findings were validated by density functional theory (DFT) calculations using the 6-31G+(d,p) method basis set implemented at Gaussian 09 program. Energy optimization calculations presented the conformational changes at the spirolactam position of CS-1 while 5hmC takes part to accommodate a probe molecule. After CS-1–5hmC complexation the energy is minimized by 19.45 kcal from the chemosensor CS-1, indicating a stable complex structure (Fig. 4 and Table S3, ESI^†). This theoretical study strongly correlates the experimental findings.Open in a separate windowFig. 4Energy diagram showing the energy differences between CS-1 and CS-1–5hmC complex.The desirable features of CS-1 such as high sensitivity and high selectivity at physiological pH encouraged us to further evaluate the potential of the chemosensor for imaging 5hmC in live cells (Fig. 5). A549 cells (Human cancer cell A549, ATCC no. CCL-185) treated with CS-1 (1 μM) exhibited weak fluorescence, whereas a deep red fluorescence signal was observed in the cells stained with CS-1 (1 μM) and 5hmC (10 μM), which is in good agreement with the FRET OFF–ON profile of the chemosensor CS-1 in presence of 5hmC, thus corroborating the in-solution observation (Fig. S14, ESI^†). Cytotoxicity assay measurement shows that the chemosensor CS-1 does not have any toxicity on the tested cells and CS-1–5hmC complex does not exert any significant adverse effect on cell viability at tested concentrations (Fig. S15, ESI^†). As far as we are aware, this is the first report where we are executing the possible use of the pyrrole–rhodamine based chemosensor for selective recognition of 5hmC in living cells. These findings open an avenue for future biomedical applications of the chemosensor to recognize 5hmC.Open in a separate windowFig. 5Confocal microscopic images of A549 cells treated with CS-1 and 5hmC. (a) Cells treated with only CS-1 at 1 μM concentration. (b) Bright field image of (a). (c) Cells treated with CS-1 and 5hmC at concentration 10 μM. (d) Bright field image of (c). All images were acquired with a 60× objective lens with the applied wavelengths: For (a) and (b), E_ex = 341 nm, E_em = 414 nm, filter used: DIDS; for (c) and (d) E_ex = 550 nm, E_em = 571 nm, filter used: Rhod-2.The concentration of 5hmC was also quantified from A549 human cancer cells. Lysate of 10⁷ A549 cells was added to 1 μM of CS-1 and the fluorescence signal was recorded. Presence of 5hmC in these cancer cells was detected with the help of CS-1–5hmC standard fluorescence curve (Fig. 6) using the selective detection ability of the chemosensor CS-1.Open in a separate windowFig. 6(a) Calibration curve obtained for the estimation of 5hmC. (b) Estimation of the concentration of 5hmC (red point) from the calibration curve.From the standard curve it was found that the concentration of 5hmC in the tested sample was 0.034 μM present in 16.7 mm³ A549 cell volume (†). Assay of 5hmC was further validated from multiple samples of A549 human cancer cells using CS-1. Increasing fold of fluorescence signals was also statistically validated after calculating the Z′ value (Table S5, ESI^†). All tested samples shows the Z′ score value more than 0.9, indicating an optimized and validated assay of 5hmC.Quantification of 5hmC in human cancer cell A549

Appraisal of novel azomethine–thioxoimidazolidinone conjugates as ecto-5′-nucleotidase inhibitors: synthesis and molecular docking studies

Purinergic signaling is regulated by a group of extracellular enzymes called ectonucleotidases. One of its members i.e., ecto-5′-nucleotidase (h-e5′NT) is involved in the final step of the enzymatic hydrolysis cascade that is the conversion of adenosine monophosphate (AMP) to adenosine and therefore, involves the regulation of adenosine level in extracellular space. The overexpression of h-e5′NT has been observed in various pathological conditions such as hypoxia, inflammation and cancers, and led to various complications. Hence, the identification of a potent as well as selective inhibitor of h-e5′NT is of greater importance in therapeutic treatment of various diseases. Azomethine-thioxoimidazolidinone derivatives were studied for their inhibition potential against e5′NT enzyme along with cytotoxic potential against cancer cell lines possessing overexpression of e5′NT enzyme. The derivative (E)-3-((4-((3-methoxybenzyl)oxy)benzylidene)amino)-2-thioxoimidazolidin-4-one (4g) displayed selective and significant inhibition towards h-e5′NT with an IC₅₀ value of 0.23 ± 0.08 μM. While two other derivatives i.e., (E)-3-(((5-bromothiophen-2-yl)methylene)amino)-2-thioxoimidazolidin-4-one (4b) and 2-thioxo-3-((3,4,5-trimethoxybenzylidene)amino)imidazolidin-4-one (4e), exhibited non-selective potent inhibitory behavior against both human and rat enzymes. Moreover, these derivatives (4b, 4e and 4g) were further investigated for their effect on the expression of h-e5′NT using quantitative real time polymerase chain reaction. Additionally, molecular docking and DFT studies were also performed to determine the putative binding mode of potent inhibitors within the enzyme active site. HOMO, LUMO, ΔE, and molecular electrostatic potential maps were computed by DFT and the charge transfer regions within the molecules were identified to find out the regions for electrophilic and nucleophilic attack.

Azomethine–thioxoimidazolidinone conjugates as ecto-5′-nucleotidase inhibitors.     

Dithieno[3,2-b:2′,3′-d]thiophene (DTT): an emerging heterocyclic building block for future organic electronic materials & functional supramolecular chemistry

Heterocyclic compounds being potent biochemical materials are ubiquitous molecules in our life. Amongst, the five membered aromatic ring systems, thiophene has emerged as a remarkable entity in organic electronics owing to its (i) high resonance energy, (ii) more electrophilic reactivity than benzene, (iii) high π-electron density, (iv) planar structure and, (v) presence of vacant d-orbital in addition to the presence of loosely bind lone-pairs of electrons on sulfur atoms. In recent past, thiophene-fused molecule namely, dithienothiophene (DTT) has attracted a tremendous attention of the researchers worldwide due to their potential applicability in organic electronics such as in solar cells, electrochromic devices (ECDs), organic field effect transistors (OFETs), organic limiting diodes (OLEDs), fluorescent probes, redox switching and so forth because of their (i) higher charge mobility, (ii) extended π-conjugation, and (iii) better tuning of band gaps, etc. In this particular review article, we envisioned to report the recent advancements made on the DTT-based architectures not only because of the potential applicability of this valuable scaffold in organic electronic but also to motivate the young researchers worldwide to look for the challenging opportunities related to this privileged building block in both material sciences and functional supramolecular chemistry.

DTT: a potential electron rich building block and its diverse application in organic electronic materials.     

Transaminase-mediated synthesis of enantiopure drug-like 1-(3′,4′-disubstituted phenyl)propan-2-amines

Transaminases (TAs) offer an environmentally and economically attractive method for the direct synthesis of pharmaceutically relevant disubstituted 1-phenylpropan-2-amine derivatives starting from prochiral ketones. In this work, we report the application of immobilised whole-cell biocatalysts with (R)-transaminase activity for the synthesis of novel disubstituted 1-phenylpropan-2-amines. After optimisation of the asymmetric synthesis, the (R)-enantiomers could be produced with 88–89% conversion and >99% ee, while the (S)-enantiomers could be selectively obtained as the unreacted fraction of the corresponding racemic amines in kinetic resolution with >48% conversion and >95% ee.

Immobilised whole-cell (R)-transaminases (TAs) enabled synthesis of either (R)- or (S)-enantiomers of drug-like amines from prochiral ketones or from racemic amines, respectively, in >95% ee.     

Synthetic strategy and structure–activity relationship (SAR) studies of 3-(5′-hydroxymethyl-2′-furyl)-1-benzyl indazole (YC-1, Lificiguat): a review

Since 1994, YC-1 (Lificiguat, 3-(5′-hydroxymethyl-2′-furyl)-1-benzylindazole) has been synthesized, and many targets for special bioactivities have been explored, such as stimulation of platelet-soluble guanylate cyclase, indirect elevation of platelet cGMP levels, and inhibition of hypoxia-inducible factor-1 (HIF-1) and NF-κB. Recently, Riociguat®, the first soluble guanylate cyclase (sGC) stimulator drug used to treat pulmonary hypertension and pulmonary arterial hypertension, was derived from the YC-1 structure. In this review, we aim to highlight the synthesis and structure–activity relationships in the development of YC-1 analogs and their possible indications.

Since 1994, YC-1 (Lificiguat) has been synthesized, and many targets for special bioactivities have been explored, such as stimulation of platelet-soluble guanylate cyclase, indirect elevation of platelet cGMP levels, and inhibition of HIF-1 and NF-κB.     

Colorimetric/photothermal dual-mode sensing detection of ascorbic acid based on a Ag[i] ion/3,3′,5,5′-tetramethylbenzidine (TMB) system

In this work, a novel strategy of colorimetric and photothermal dual-mode sensing determination of ascorbic acid (AA) based on a Ag⁺/3,3′,5,5′-tetramethylbenzidine (TMB) system was developed. In this sensing system, Ag⁺ could oxidize TMB with a distinct color change from colorless to blue color, strong absorbance at 652 nm and a photothermal effect under 808 nm laser irradiation due to the formation of oxidized TMB (oxTMB). When AA was present, oxTMB was reduced accompanied by a change from blue to colorless, and a decrease in absorption peak intensity and the photothermal effect. AA concentration showed a negative linear correlation with the value of both the absorbance intensity at 652 nm and temperature in the range of 0.2–10 μM (A = −0.03C + 0.343 (R², 0.9887; LOD, 50 nM); ΔT = −0.57C + 8.453 (R², 0.997; LOD, 7.8 nM)). Based on this, a sensing approach for detection of AA was proposed with dual-mode and without the complicated synthesis of nanomaterials. The photothermal effect and colorimetric signal provided a dual-mode detection strategy for AA, overcoming the limitations of any single mode. This colorimetric and photothermal dual-mode detection has great potential in the detection of AA in clinical pharmaceuticals and the construction of portable and highly sensitive sensors.

Colorimetric and photothermal dual-readout sensing detection of AA using a Ag⁺/TMB system.     

Computational studies on functionalized Janus MXenes MM′CT2, (M,M′ = Zr,Ti, Hf,M ≠ M′; T = –O, –F, –OH): photoelectronic properties and potential photocatalytic activities

Motivated by the successful synthesis of Janus monolayers of transition metal dichalcogenides (i.e., MoSSe), we computationally investigated the structural, electronic, optical, and transport properties of functionalized Janus MXenes, namely MM′CT₂ (M, M′ = Zr, Ti, Hf, M ≠ M′, T = –O, –F, –OH). The results of the calculations demonstrate that five stable O-terminated Janus MXenes (ZrTiCO₂-I, ZrHfCO₂-I, ZrHfCO₂-III, HfTiCO₂-I, and HfTiCO₂-III), exhibit modest bandgaps of 1.37–1.94 eV, visible-light absorption (except for ZrHfCO₂-I), high carrier mobility, and promising oxidization capability of photoinduced holes. Additionally, their indirect-gap, spatially separated electron–hole pairs, and the dramatic difference between the mobilities of electrons and holes could significantly limit the recombination of photoinduced electron–hole pairs. Our results indicate that the functionalized Janus MXene monolayers are ideal and promising materials for application in visible light-driven photocatalysis.

The functionalized Janus MXene monolayers: ideal and promising materials for use in visible light-driven photocatalysis.     

Metal-free oxidative coupling of arylmethylamines with indoles: a simple,environmentally benign approach for the synthesis of 3,3′-bis(indolyl)methanes

The efficient metal-free oxidative coupling of arylmethylamines with indoles has been developed using molecular oxygen as a green oxidant. The present reaction provides a novel route towards the synthesis of 3,3′-bis(indolyl)methanes in excellent yields of up to 95% via C–C and C–N bond formation. This attractive and environmentally friendly one-pot protocol is a simple procedure that features inexpensive acetic acid as the catalyst and molecular oxygen as the sole oxidant, and it supports a wide substrate scope with the good tolerance of functional groups.

The efficient metal-free oxidative coupling of arylmethylamines with indoles has been developed using molecular oxygen as a green oxidant towards a novel rout of 3,3′-bis(indolyl)methanes (BIMs) synthesis.