Discovery of a small-molecule inhibitor and cellular probe of Keap1–Nrf2 protein–protein interaction

A high-throughput screen (HTS) of the MLPCN library using a homogenous fluorescence polarization assay identified a small molecule as a first-in-class direct inhibitor of Keap1–Nrf2 protein–protein interaction. The HTS hit has three chiral centers; a combination of flash and chiral chromatographic separation demonstrated that Keap1-binding activity resides predominantly in one stereoisomer (SRS)-5 designated as ML334 (LH601A), which is at least 100× more potent than the other stereoisomers. The stereochemistry of the four cis isomers was assigned using X-ray crystallography and confirmed using stereospecific synthesis. (SRS)-5 is functionally active in both an ARE gene reporter assay and an Nrf2 nuclear translocation assay. The stereospecific nature of binding between (SRS)-5 and Keap1 as well as the preliminary but tractable structure–activity relationships support its use as a lead for our ongoing optimization     

Methodical considerations when estimating nutrient digestibility in horses using the mobile bag technique

Total collection of faeces is considered the golden standard for estimating apparent total tract digestibility (ATTD) in horses. However, the evaluation of individual feedstuffs is limited and determination of nutrient digestibility in different segments of the gastrointestinal tract (GIT) is excluded. The rationale for performing this study was that the mobile bag technique (MBT) can provide information on individual feedstuffs' degradation, and the use of fistulated animals does provide additionally information regarding degradation in individual segments of the GIT. Recommendations for using the MBT in ruminants are well established, but limited methodical studies have been published with horses. The objective of this study was to evaluate the MBT by comparing the ATTD with the nutrient disappearance and degradation kinetics of hay in horses. It was hypothesised that DM degradation as estimated by the MBT is equal to the ATTD of the DM. Furthermore, we hypothesised that bag size has no effect on nutrient disappearance but increasing the feed to surface area (FSA) decreases the DM disappearance. Five caecum cannulated horses were fed a hay-only diet (6.7 kg DM/day) with 14 days of adaptation followed by four consecutive days of total faeces collection. Three bag sizes (height × length × side, cm; 1.2 × 10 × 2, 3 × 4 × 2, 1 × 6 × 2) and three FSAs (10.4, 20.8 and 41.7 mg/cm²) were administrated at each meal (3 meals/day) on days 1 and 2 of the collection. Faeces were checked for bags every 6th h, the collection time was noted and the DM disappearance together with the transit time (TT) for each bag type was estimated. Dry matter disappearance from the individual bags was fitted to degradation profiles, and the effective degradability (ED) and degradation (D_t) were determined. The results of the study showed that the ATTD of DM, organic matter (OM), NDF and ADF can be predicted based on their disappearance from the mobile bags, but that ash and CP are overestimated in comparison to the ATTD. The TT for the bags was 29.2 h, and when using a mean retention time of 30 h to predict ED and D_t, it was clear that ED was underestimated, whereas D_t reflected the ATTD of DM. In conclusion, the MBT can be used to estimate the degradability of DM, OM and fibre as these nutrients resemble the ATTD. The bag size did not affect the DM disappearance, but the FSA should be kept below 20 mg/cm² as higher levels might limit the degradation kinetics.     

New type of arrangement of rare-earth quinolinolate. Molecular structure of scandium 2-methyl-8-quinolinolate

The tetrakis 2-methyl-8-quinolinolate scandium complex [Sc(q^Me)₄(H)] (1) have been prepared by the reaction of ScCl₃ with 2-methyl-8-quinolinol (Hq^Me) in methanol in the presence of aqueous ammonia. The X-ray diffraction analysis has shown that a molecule of (1) has a propeller like shape herein the Sc(III) ion is surrounded by four methylquinolinolate ligands two of which are chelate but the other two are monodentate and one of these monodentate methylquinolinolate ligands contains hydrogen atom at nitrogen atom. Furthermore there are two molecules of water per one complex molecule not coordinated to the Sc cation that is not typical for rare-earth compounds.     

Glutamine synthetase from Salmonella typhimurium: manganese(II), substrate, and inhibitor interaction with the unadenylylated enzyme.

Unadenylylated glutamine synthetase (EC 6.3.1.2) was isolated and purified to homogeneity from Salmonella typhimurium. The enzyme molecule is a symmetrical aggregate of 12 subunits arranged in two hexagonal layers, as is evident from electron micrographs. The subunit molecular weight of the enzyme was found to be approximately 50,000 by polyacrylamide gel electrophoresis in sodium dodecyl sulfate when compared to Escherichia coli glutamine synthetase and other protein standards. A long tube of glutamine synthetase was formed as a single-stranded coil resulting from incubation of the enzyme in a low ionic strength buffer. A study of Mn(II) binding to the unadenylylated enzyme at 25 °C was conducted as a function of pH. At pH 7.1 two classes of metal ion sites per subunit were found with K_D values of 3.7 × 10^?6 and 1.7 × 10^?4m, while at pH 6.8 these values were 1.1 × 10^?5 and 1.0 × 10^?4m, respectively. Only one set of binding sites was observed at pH 6.2 with a K_D value of 1.0 × 10^?4m. The metal ion binding sites were further investigated by monitoring proton relaxation rates (prr) and the epr spectrum of enzyme-bound Mn(II). The longitudinal prr of water protons at pH 7.1 indicate that protons interacting with enzyme-Mn(II) at the “tight” site (K_D = 3.7 × 10^?6) are de-enhanced (?_b1 = 0.42) and result from water protons beyond the inner coordination sphere. The second Mn(II) site has a value of ?_b2 = 35 for the binary enhancement, suggesting that this site probably has two to three rapidly exchanging water molecules in its coordination sphere. The epr spectrum of enzyme-bound Mn(II) at the “tight” site is isotropic and is dramatically sharpened by adding the substrate analog methionine sulfoximine. Subsequent addition of ATP or the ATP analog, AMP-PCP (adenylyl methylene diphosphate) produced anisotropic spectra that were similar, suggesting that both ATP and AMP-PCP bind similarly on the enzyme surface. However, a marked change in the Mn(II) environment from anisotropic to near cubic results from the addition of ADP to the quaternary enzyme-Mn(II)-sulfoximine- (AMP-PCP) complex, indicating that ADP displaces AMP-PCP. No change in the anisotropic spectrum due to the enzyme-Mn(II)-sulfoximine-ATP complex is seen by the addition of ADP. This experimental result supports the experimental findings of Ronzio and Meister [Proc. Nat. Acad. Sci. USA59, 164 (1968)], who established that ATP phosphorylates methionine sulfoximine, thereby producing an inactive enzyme. The allosteric effectors, AMP and Trp, have little effect on the epr spectrum of the complex formed from Mn(II), enzyme, sulfoximine, and ADP, suggesting the absence of direct coordination of AMP or Trp to the bound Mn(II). The prr and epr results reported herein with glutamine synthetase from S. typhimurium when compared to those seen for the enzyme from E. coli [Villafranca et al., Biochemistry15, 544 (1976)] demonstrate some similarities but also many substantial differences between the enzymes from these two bacterial sources.     

Synthesis, characterization, and reactivity of zinc carboxylate complexes of 2,3-pyridine dicarboxylic acid and (3-oxo-2,3-dihydro-benzo[1,4]oxazin-4-yl)acetic acid

Ethylenediammonium tris-2,3-pyridine dicarboxylato zinc(II) trihydrate (enH₂)₂[ZnL₃]·2H₂O (1) (where H₂L = 2,3-pyridine dicarboxylic acid, en = ethylenediamine) and a mixed metal coordination polymer with composition [Na₂ZnL′₃(OAc)(H₂O)₃]_n (2) {where L′ = anion of (3-oxo-2,3-dihydro-benzo[1,4]oxazin-4-yl)acetic acid} are synthesized and characterized. The complex 1 is mono nuclear complex with three coordinating carboxylate anion along with nitrogen chelating zinc ion and there is three uncoordinated carboxylate group one each from three ligand molecules making a complex anion of zinc(II). The zinc(II) ion are in distorted octahedral coordination geometry. In this complex diprotonated ethylenediamines serve as cations. The complex 2 has a polymeric structure with one acetate and three carboxylate of L′ binding to zinc ion provides a tetrahedral environment and these ligands further hold dinuclear units of tri-aquated sodium ions; each dinuclear sodium units are bridged by one water molecule make the coordination polymer. The catalytic ability of these two complexes 1 and 2 towards carbon-carbon bond formation reaction between 3,4-dimethoxy benzaldehyde and acetone are studied. Both the complexes as well as sodium salt of L′ are found to be catalyst for such reactions.     

Determination of the molecular weight of proteins in heterogeneous mixtures: use of an air-driven ultracentrifuge for the analysis of protein--protein interactions.

A high-speed air-driven ultracentrifuge (Airfuge) has been used to study the molecular weights of proteins in heterogeneous mixtures. The method is based on previous studies (M. A. Bothwell, G. J. Howlett, and H. K. Schachman, 1978, J. Biol. Chem., 253, 2073–2077) which showed that at sedimentation equilibrium in the Airfuge the fraction of a protein remaining in an upper fraction of the Airfuge tube is almost linearly related to the exponential of the reduced molecular weight of the protein. In this study the total fraction of each particular protein remaining in an upper fraction of the Airfuge tube is determined by quantitative sodium dodecyl sulfate-gel electrophoresis. This procedure allows a wide range of proteins to be analyzed in a single Airfuge experiment. The method yields the “native” molecular weights of the protein components and is independent of the shape of the macromolecules being studied. Interactions occurring between the components in solution can be detected from the Airfuge data, and procedures are described which allow the experimental data for such interactions to be analyzed in terms of an equilibrium constant for the interaction. Results obtained for the electrostatic interaction at neutral pH between lysozyme and ovalbumin (K = 1.1 × 10⁵, m^?1) and lysozyme and bovine serum albumin (K = 1.0 × 10⁵, m^?1) agree well with literature values.     

A new mode of B12 binding and the direct participation of a potassium ion in enzyme catalysis: X-ray structure of diol dehydratase

Background: Diol dehydratase is an enzyme that catalyzes the adenosylcobalamin (coenzyme B₁₂) dependent conversion of 1,2-diols to the corresponding aldehydes. The reaction initiated by homolytic cleavage of the cobalt–carbon bond of the coenzyme proceeds by a radical mechanism. The enzyme is an α₂β₂γ₂ heterooligomer and has an absolute requirement for a potassium ion for catalytic activity. The crystal structure analysis of a diol dehydratase–cyanocobalamin complex was carried out in order to help understand the mechanism of action of this enzyme.Results: The three-dimensional structure of diol dehydratase in complex with cyanocobalamin was determined at 2.2 Å resolution. The enzyme exists as a dimer of heterotrimers (α β γ)₂. The cobalamin molecule is bound between the α and β subunits in the ‘base-on’ mode, that is, 5,6-dimethylbenzimidazole of the nucleotide moiety coordinates to the cobalt atom in the lower axial position. The α subunit includes a (β/α)₈ barrel. The substrate, 1,2-propanediol, and an essential potassium ion are deeply buried inside the barrel. The two hydroxyl groups of the substrate coordinate directly to the potassium ion.Conclusions: This is the first crystallographic indication of the ‘base-on’ mode of cobalamin binding. An unusually long cobalt–base bond seems to favor homolytic cleavage of the cobalt–carbon bond and therefore to favor radical enzyme catalysis. Reactive radical intermediates can be protected from side reactions by spatial isolation inside the barrel. On the basis of unique direct interactions between the potassium ion and the two hydroxyl groups of the substrate, direct participation of a potassium ion in enzyme catalysis is strongly suggested.     

NOTA analogue: A first dithiocarbamate inhibitor of metallo-β-lactamases

The emergence of antibiotic drug (like carbapenem) resistance is being a global crisis. Among those resistance factors of the β-lactam antibiotics, the metallo-β-lactamases (MBLs) is one of the most important reasons. In this paper, a series of cyclic dithiocarbamate compounds were synthesized and their inhibition activities against MBLs were initially tested combined with meropenem (MEM) by in vitro antibacterial efficacy tests. Sodium 1,4,7-triazonane-1,4,7-tris(carboxylodithioate) (compound 5) was identified as the most active molecule to restore the activity of MEM. Further anti-bacterial effectiveness assessment, compound 5 restored the activity of MEM against Escherichia coli, Citrobacter freundii, Proteus mirabilis and Klebsiella pneumonia, which carried resistance genes of bla_NDM-1. The compound 5 was non-hemolytic, even at a concentration of 1000?µg/mL. This compound was low toxic toward mammalian cells, which was confirmed by fluorescence microscopy image and the inhibition rate of HeLa cells. The Ki value of compounds 5 against NDM-1 MBL was 5.63?±?1.27?μM. Zinc ion sensitivity experiments showed that the inhibitory effect of compound 5 as a MBLs inhibitor was influenced by zinc ion. The results of the bactericidal kinetics displayed that compound 5 as an adjuvant assisted MEM to kill all bacteria. These data validated that this NOTA dithiocarbamate analogue is a good inhibitor of MBLs.     

Synthesis and biological evaluation of polyhydroxylated oxindole derivatives as potential antileishmanial agent

The devastating appearance of numerous drug-unresponsive strains of Leishmania donovani and severe toxic side effects of conventional antileishmanial therapy necessitates the search for novel leads, to treat visceral leishmaniasis efficiently. The current study deals with the synthesis and biological evaluation of a unique C-5 functionalized oxindole based polyphenol to ascertain its activities against L. donovani infection, in vitro. The polyhydroxylated oxindole derivative (1) was generated by coupling styrene derivatives with 5-bromo bis-arylidene oxindole using Heck coupling reaction. The synthesized molecule 1 was tested for its antileishmanial activity using both promastigote and amastigote stages of L. donovani. Molecule 1 showed promising anti-promastigote and anti-amastigote activities with IC₅₀ values 15?µM and 1?µM, respectively, with no cytotoxicity towards host splenocytes. The results revealed that this compound induced parasite death by promoting oxidative stress, thereby triggering apoptosis.     

Energy and protein requirements of Holstein × Gyr crossbred heifers

Nutrient requirements in cattle are dependent on physiological stage, breed and environmental conditions. In Holstein × Gyr crossbred dairy heifers, the lack of data remains a limiting factor for estimating energy and protein requirements. Thus, we aimed to estimate the energy and protein requirements of Holstein × Gyr crossbred heifers raised under tropical conditions. Twenty-two crossbred (½ Holstein × ½ Gyr) heifers with an average initial BW of 102.2 ± 3.4 kg and 3 to 4 months of age were used. To estimate requirements, the comparative slaughter technique was used: four animals were assigned to the reference group, slaughtered at the beginning of the experiment to estimate the initial empty BW (EBW) and composition of the animals that remained in the experiment. The remaining animals were randomized into three treatments based on targeted rates of BW gain: high (1.0 kg/day), low (0.5 kg/day) and close to maintenance (0.1 kg/day). At the end of the experiment, all animals were slaughtered to determine EBW, empty body gain (EBG) and body energy and protein contents. The linear regression parameters were estimated using PROC MIXED of SAS (version 9.4). Estimates of the parameters of non-linear regressions were adjusted through PROC NLIN of SAS using the Gauss–Newton method for parameter fit. The net requirements of energy for maintenance (NE_m) and metabolizable energy for maintenance (ME_m) were 0.303 and 0.469 MJ/EBW^0.75 per day, respectively. The efficiency of use of ME_m was 64.5%. The estimated equation to predict the net energy requirement for gain (NE_g) was: NE_g (MJ/day) = 0.299 × EBW^0.75 × EBG^0.601. The efficiency of use of ME for gain (k_g) was 30.7%. The requirement of metabolizable protein for maintenance was 3.52 g/EBW^0.75 per day. The equation to predict net protein requirement for gain (NP_g) was: NP_g (g/day) = 243.65 × EBW^−0.091 × EBG. The efficiency of use of metabolizable protein for gain (k) was 50.8%. We observed noteworthy differences when comparing to ME and protein requirements of Holstein × Gyr crossbred heifers with other systems. In addition, we also observed differences in estimates for NE_m, NE_g, NP_g, k_g and k. Therefore, we propose that the equations generated in the present study should be used to estimate energy and protein requirements for Holstein × Gyr crossbred dairy heifers raised in tropical conditions in the post-weaning phase up to 185 kg of BW.     

Synthesis of 1-benzyl-1H-benzimidazoles as galectin-1 mediated anticancer agents

In our pursuit to develop novel non-carbohydrate small molecule Galectin-1 Inhibitors, we have designed a series of 1-benzyl-1H-benzimidazole derivatives and demonstrated their anticancer activity. The compound 6g, 4-(1-benzyl-5-chloro-1H-benzo[d]imidazol-2-yl)-N-(4-hydroxyphenyl) benzamide was found to be most potent with an IC₅₀ of 7.01 ± 0.20 µM and arresting MCF-7 cell growth at G2/M phase and S phase. Induction of apoptosis was confirmed by morphological changes like cell shrinkage, blebbing and cell wall deformation, dose dependent increase in the mitochondrial membrane potential (ΔΨm) and ROS levels. Further, dose dependent decrease in Gal-1 protein levels proves Gal-1 mediated apoptosis by 6g. Molecular docking studies were performed to understand the Gal-1 interaction with compound 6g. In addition, RP-HPLC studies showed 85.44% of 6g binding to Gal-1. Binding affinity studies by fluorescence spectroscopy and Surface Plasmon Resonance (SPR) showed that 6g binds to Gal-1 with binding constant (K_a) of 1.2 × 10⁴ M⁻¹ and equilibrium constant K_D value of 5.76 × 10⁻⁴ M respectively.     

Metal-ion-catalyzed hydrolysis of monomethyl and dimethyl esters of 3-(2-imidazoleazo)phthalic acid: Bifunctional catalysis by carboxyl group and the metal ion in the hydrolysis of an alkyl ester

The Cu(II) or Ni(II) ion-catalyzed hydrolysis of methyl 2-carboxy-6-(2-imidazoleazo)benzoate (1) and the corresponding dimethyl ester (2) was studied kinetically at various pH values. For 2, the ester group located at the o position to the azo substiuent was hydrolyzed. From the rate data obtained at various metal concentrations, the values of k_cat and K_f were estimated at each pH value. For the Ni(II)-catalyzed hydrolysis of 1 at pH < 4, k_cat increases as pH is lowered, indicating bifunctional catalysis by the carboxyl group and the metal ion. For most of the reactions investigated under other conditions, the ester hydrolysis was subjected to sole catalysis by the metal ions. Detailed analysis of kinetic data obtained for these reactions indicated that the metal-ion catalysis involves the rate-determining breakdown of the tetrahedral intermediates formed by the addition of a water molecule or hydroxide ion. The bifunctional catalysis by the carboxyl group and Ni(II) ion can be considered as a model for carboxypeptidase A. The kinetic data indicate that the bifunctional catalysis proceeds through the nucleophilic attack of the carboxylate ion at the Ni(II)-coordinated carbonyl group.     

Metabolizable amino acids and energy requirements of Nellore and crossbred Angus × Nellore bulls fed rations of different crude protein concentrations

Growth rate of cattle depends on their genetic makeup and nutrient intake. Moreover, increased growth rate may lead to increased amino acid (AA) requirements. Therefore, we evaluated the AA content of the empty body and estimated the net AA and energy requirements of purebred and crossbred beef bulls fed rations of different dietary CP concentrations. We performed a comparative slaughter experiment with 24 Nellore and 24 Angus × Nellore (A × N) bulls (8 months; initial shrunk BW: Nellore = 208.0 ± 12.78 kg; A × N = 221.9 ± 14.16 kg). Eight bulls (four Nellore and four A × N) were designated as the reference group, eight bulls (four Nellore and four A × N) were fed to maintenance level and 32 bulls (16 Nellore and 16 A × N) were fed ad libitum. The 32 bulls fed ad libitum were distributed using a completely randomized design in a 2 × 3 factorial scheme with two genetic groups (Nellore or A × N) and three dietary CP contents (100, 120 or 140 g CP/kg DM), being four groups with five bulls and two groups with six bulls. The experimental period lasted for 224 days. There were no interactions (P ≥ 0.056) between the dietary CP contents and genetic groups for any of the response variables. The dietary CP contents did not affect (P ≥ 0.062) the AA content in the empty body (g/kg empty BW [EBW]), with exception for Tryptophan (P = 0.027, linear effect). The dietary CP contents did not affect (P ≥ 0.051) AA content in the empty body (g/100 g of CP), with exception for Alanine (P = 0.013) that responded quadratically to dietary CP increase. The equations to estimate the net Lysine (Lys) and Methionine (Met) requirements (g/100 g of CP) were: Lys = 5.1 × EBW^0.0594 and Met = 1.7 × EBW^0.0255. Metabolizable Lys and Met to metabolizable energy (ME) ratios decreased as bulls EBW increased. Also, the metabolizable protein to ME ratio decreased as bulls EBW increased. In conclusion, the present study provides useful information regarding net and metabolizable requirements of AA of purebred and crossbred beef bulls. In the future, after the validation of the equations, these results can be used to calculate the AA requirements for growth of purebred and crossbred beef bulls. Nevertheless, it is important to highlight that the small sample size was one limitation of this present experiment.     

Comparison of nonlinear models to describe the feather growth and development curve in yellow-feathered chickens

Feathers play a critical role in thermoregulation and directly influence poultry production. Poor feathering adversely affects living appearance and carcass quality, thus reducing profits. However, producers tend to ignore the importance of feather development and do not know the laws of feather growth and development. The objective of this study was to fit growth curves to describe the growth and development of feathers in yellow-feathered broilers during the embryonic and posthatching periods using different nonlinear functions (Gompertz, logistic and Bertalanffy). Feather mass and length were determined during the embryonic development and posthatching stages to identify which growth model most accurately described the feather growth pattern. The results showed that chick embryos began to grow feathers at approximately embryonic (E) day 10, and the feathers grew rapidly from E13 to E17. There was little change from E17 to the day of hatching (DOH). During the embryonic period, the Gompertz function (Y = 798.48e^{−203 431exp(−0.87t)}, Akaike’s information criterion (AIC) = −0.950 × 10³, Bayesian information criterion (BIC) = −0.711 × 10³ and mean square error (MSE) = 559.308) provided the best fit for the feather growth curve compared with the other two functions. After hatching, feather mass and length changed little from the DOH to day (D) 14, increased rapidly from D21 to D91 and then grew slowly after D91. The first stage of feather molting occurred from 2 to 3 weeks of age when the down feathers were mostly shed and replaced with juvenile feathers, and the second stage occurred at approximately 13 to 15 weeks of age. The three nonlinear functions could overall fit the feather growth curve well, but the Bertalanffy model (Y = 116.88 × (1−0.86e^−0.02t)³, AIC = 1.065 × 10⁵, BIC = 1.077 × 10⁵ and MSE = 11.308) showed the highest degree of fit among the models. Therefore, the Gompertz model exhibited the best goodness of fit for the feather growth curve during the embryonic development, while the Bertalanffy model was the most suitable model due to its accurate ability to predict the growth and development of feathers during the growth period, which is an important commercial characteristic of yellow-feathered chickens.     

Synthesis,X-ray crystal structure study and antitumoral evaluations of 5,6-disubstituted pyrimidine derivatives

5,6-Disubstituted pyrimidine derivatives (3–20) were prepared by intramolecular cyclization reaction of α-(1-carbamyliminomethylene)-γ-butyrolactone (2) with sodium ethoxide and subsequent chemical transformation of 2-hydroxy group in C-5 side chain as well as lithiation reaction for introduction of acyclic side chain at C-6. All compounds were characterized by ¹H NMR, ¹³C NMR and mass spectra. Structures of compounds 4, 7 and 14 were unambiguously confirmed by X-ray crystal structural analysis. Supramolecular structures of these three compounds differ significantly. Two N–H?O and one C–H?O hydrogen bonds in 4 form three-dimensional network. One O–H?N hydrogen bond and one π?π interaction self-assemble the molecules of 7 into sheets. In supramolecular aggregation of 14, only π?π stacking interactions participate, so forming chains. The compounds were evaluated for their cytostatic activities against human malignant cell lines. Of all tested compounds, 2,4-dimethoxy-5-methoxytritylethylpyrimidine (9) and 2,4-dichloro-5-chloroethylpyrimidine (14) exhibited the most prominent inhibitory effects. Furthermore, compound 14 showed marked activity against human colon carcinoma (IC₅₀ = 0.4 μM).     

Aggregation of tyrocidine in aqueous solutions.

The formation of aggregates of tyrocidine B at 4°C and 20°C in aqueous solutions was studied by means of light scattering and fluorescent techniques. The apparent weight molecular weight of tyrocidine B aggregates was found to be 36,000 at 4°C and 28,800 at 20°C. Fluorescence titration experiments with dansyl-chloride resulted in an aggregational number of 31 (4°) and 28 (20°) indicating that one molecule of dye is bound per monomer of molecular weight 1,200. From a Scatchard plot apparent association constants of 1.22 × 10⁵ M (4°) and 0.95 × 10⁵ M (20°) were calculated. From the angular dependence of scattered intensity the radii of gyration were determined to be 60 Å and 58 Å, respectively.     

Cytotoxicity and DNA binding property of phenanthrene imidazole with polyglycol side chains

A series of phenanthrene imidazole with polyglycol side chain (2a–2c and 3a–3c) were synthesized and characterized by IR, NMR and MS. The cytotoxicity of 2a–2c and 3a–3c against cancer cell lines (HL-60, BGC-823, Bel-7402 and KB) in vitro were measured using MTT method. The DNA binding properties of 3a–3c were investigated by UV, fluorescence, CD spectroscopies and thermal denaturation. The results indicate that 2a exhibits higher cytotoxicity than cisplatin against BGC-823 and Bel-7402 cell lines, 3b and 3c exhibit higher cytotoxicity than 2b and 2c against BGC-823, Bel-7402 and KB cell lines. The cytotoxic effect of 2a–2c decrease with the increase of side chains length, the cytotoxic effect of 3a–3c increased with the increasing length of side chains against BGC-823, Bel-7402 and KB cell lines. Compounds 3a–3c intercalated DNA with a vertical orientation in the intercalation pocket. The binding constants of 3a–3c with Ct-DNA are 1.68 × 10⁶, 1.51 × 10⁶ and 0.709 × 10⁶ M^?1, respectively. The binding affinity of 3a–3c with Ct-DNA trended to decrease with the increasing length of polyglycol side chains.     

The kinetics and mechanism of oxidation of maltose and lactose by Tl(III) in acidic media

The kinetics of the cleavage of d-arabino-hexos-2-ulose (1) and of glyoxal (2) with hydrogen peroxide in alkaline water and in 44% (w/w) ethanol-water solutions (pOH 0.5-5) were studied over a temperature range of ?25 to +25°. The relative rate of the competing reactions of 1 with the cleavage in 0.03-1M sodium hydroxide was determined from the rate of formation of hydrogen peroxide in the oxidation of d-glucose to 1 with 2-anthraquinonesulfonic acid in the presence of oxygen at 25 and 40°. The cleavages of both 1 and 2 were first-order with respect to hydrogen peroxide, and also to hydroxyl ion at low alkalinities. The rate of cleavage of 1 reached a maximum at pOH ～2.5, whereas the competing reactions of 1 and the cleavage of 2 were constantly accelerated with increasing hydroxyl-ion concentration. Unlike 2, compound 1 was cleaved more rapidly in ethanol-water than in water. The activation energies of the cleavage of 1 and 2, and the competing reactions of 1, were 49, 57, and 65 kJ.mol^?1, respectively.     

Synthesis, characterization, crystal structures and magnetic properties of dissymmetrical double schiff base heterotrinuclear complexes: [CuL(H2O)CoCuL] · H2O · CH3OH and [(CuL)2Ni] · 2H2O

A dissymmetrical double Schiff base Cu(II) mononuclear complex: CuHL (1) (where H₃L is N-3-carboxylsalicylidene-N^′-salicylaldehyde-1,2-diaminoethane) and two trinuclear complexes: [CuL(H₂O)CoCuL] · H₂O · CH₃OH (2) and [(CuL)₂Ni] · 2H₂O (3) have been synthesized and characterized by means of elemental analyses, IR and electronic spectra. The crystal structures of two heterotrinucler complexes were determined by X-ray analysis. Each dissymmetrical cell unit of the complex 2 contains two heterotrinucler neutral molecules. In each neutral molecule, the central Co²⁺ ion is located at the site of O₆ with a distorted octahedral geometry and one terminal Cu²⁺ ion at the four-coordination site of N₂O₂, but the other one at the square-pyramidal environment of N₂O₃. Each dissymmetrical unit of the complex 3 contains a heterotrinucler neutral molecule, whose structure is similar to that of 2 except two terminal Cu²⁺ ions both at the inner site of N₂O₂. The magnetic properties of two heterotrinucler complexes have been determined in the temperature range of 5-300 K, which indicate that the interaction between the central Co²⁺ ion or Ni²⁺ ion and the outer Cu²⁺ ions is antiferromagnetic. The exchange integrals are equal to −26.2 cm⁻¹ for 2 and −50.6 cm⁻¹ for 3.