Structure-based design of Aurora A & B inhibitors

The Aurora family of serine/threonine kinases are mitotic regulators involved in centrosome duplication, formation of the bipolar mitotic spindle and the alignment of the chromosomes along the spindle. These proteins are frequently overexpressed in tumor cells as compared to normal cells and are therefore potential therapeutic oncology targets. An Aurora A high throughput screen revealed a promising sub-micromolar indazole-benzimidazole lead. Modification of the benzimidazole portion of the lead to a C2 linker with a phenyl ring was proposed to achieve novelty. Docking revealed that a conjugated linker was optimal and the resulting compounds were equipotent with the lead. Further structure-guided optimization of substituents on the 5 & 6 position of the indazole led to single digit nanomolar potency. The homology between the Aurora A & Aurora B kinase domains is 71% but their binding sites only differ at residues 212 & 217 (Aurora A numbering). However interactions with only the latter residue may be used for obtaining selectivity. An analysis of published Aurora A and Aurora B X-ray structures reveals subtle differences in the shape of the binding sites. This was exploited by introduction of appropriately sized substituents in the 4 & 6 position of the indazole leading to Aurora B selective inhibitors. Finally we calculate the conformational energy penalty of the putative bioactive conformation of our inhibitors and show that this property correlates well with the Aurora A binding affinity.     

Coupling of CpCr(CO)3 and heterocyclic dithiadiazolyl radicals. synthetic, x-ray diffraction, dynamic NMR, EPR, CV, and DFT studies

The reaction of the 1,2,3,5-dithiadiazolyls (4-R-C(6)H(4)CN(2)S(2))(2) (R = Me, 2a; Cl, 2b; OMe, 2c; and CF3, 2d) and (3-NC-5-tBu-C(6)H(3)CN(2)S(2))(2) (2e) with [CpCr(CO)(3)](2) (Cp = eta(5)-C(5)H(5)) (1) at ambient temperature respectively yielded the complexes CpCr(CO)(2)(eta(2)-S(2)N(2)CC(6)H(4)R) (R = 4-Me, 3a; 4-Cl, 3b; 4-OMe, 3c; and 4-CF(3), 3d) and CpCr(CO)(2)(eta(2)-S(2)N(2)CC(6)H(3)-3-(CN)-5-(tBu)) (3e) in 35-72% yields. The complexes 3c and 3d were also synthesized via a salt metathesis method from the reaction of NaCpCr(CO)(3) (1B) and the 1,2,3,5-dithiadiazolium chlorides 4-R-C(60H(4)CN(2)S(2)Cl (R = OMe, 8c; CF(3), 8d) with much lower yields of 6 and 20%, respectively. The complexes were characterized spectroscopically and also by single-crystal X-ray diffraction analysis. Cyclic voltammetry experiments were conducted on 3a-e, EPR spectra were obtained of one-electron-reduced forms of 3a-e, and variable temperature 1H NMR studies were carried out on complex 3d. Hybrid DFT calculations were performed on the model system [CpCr(CO)(2)S(2)N(2)CH] and comparisons are made with the reported CpCr(CO)(2)(pi-allyl) complexes.     

Synthetic and X-ray structural and reactivity studies of Cp*RuIV complexes containing bidentate dithiocarbonate, xanthate, carbonate, and phosphinate ligands (Cp* = eta5-C5Me5)

The reaction of [Cp*RuCl2]2 (1; Cp* = eta5-C5Me5) with tetraalkyldithiuram disulfides (R2NC(S)SS(S)CNR2, R = Me, Et), isopropylxanthic disulfide ([iPrOC(S)S]2), and bis(thiophosphoryl) disulfide ([(iPrO)2P(S)S]2) led to the isolation of dark-red crystalline solids of Cp*RuIVCl2(eta2-dithiolate) complexes [dithiolate = S2CNR2, DTCR (2a, R = Me; 2b, R = Et), S2COiPr (3), and S2P(iPrO)2 (4)]. Dichlorido substitution in 2 and 3 with DTCEt and S2COiPr anions yielded RuIV derivatives containing bis(DTC) and mixed DTC-dithiocarbonate ligands. These are the first organoruthenium complexes of such ligands. The reaction of monophosphines with 2a resulted in monochlorido substitution, whereas the analogous reaction with 3 resulted in displacement of both chlorido ligands and reduction of the metal center to RuII. Reduction at Ru was also observed in the reaction of 2a with [CpCr(CO)3]2. Of these complexes, only 2 and 3 are air-stable in the solid state for an extended period. All of the complexes have been spectrally characterized, and selected compounds are also crystallographically characterized.     

Polyamide-imide/polyetherimide dual-layer hollow fiber membranes for pervaporation dehydration of C1–C4 alcohols

To circumvent the common swelling and deteriorated performance of integral asymmetric hollow fiber membranes for pervaporation dehydration, we have developed novel polyamide-imide (PAI)/polyetherimide (PEI) hollow fiber membranes with synergized performance with the aid of dual-layer spinning technology. Dehydration of C1–C4 alcohols has been conducted and the orders of their fluxes and permeances have been analyzed. The hollow fibers spun at 2 cm air gap and annealed at 75 °C exhibit the highest pervaporation performance: separation factors for t-butanol/water and iso-butanol/water binary systems are greater than 50,000 with flux more than 700 g/m² h. A comparison with literature data shows that the newly developed membranes outperform most other polymeric membranes for the dehydration of IPA and butanols. The dual-layer hollow fiber membranes also exhibit good long-term stability up to 200 h. The superior performance can be attributed to (1) the balanced properties of PAI as the selective layer for dehydration pervaporation; (2) the low water uptake and less swelling characteristic of the PEI supporting layer; and (3) the desirable membrane morphology consisting of a fully porous inner layer, a porous interface, and an ultrathin dense-selective outer skin.     

Determination of senkirkine and senecionine in Tussilago farfara using microwave-assisted extraction and pressurized hot water extraction with liquid chromatography tandem mass spectrometry

Tussilago farfara (Kuan Donghua) is an important Chinese herbal medicine which has been shown to contain many bioactive compounds and widely used to relieve cough and resolve phlegm. However, besides therapeutic bioactive compounds, this herb has been found to contain toxic pyrrolizidine alkaloids (PAs), mainly senkirkine and traces of senecionine. In this report, conditions for microwave-assisted extraction (MAE) and pressurized hot water extraction (PHWE) were optimized for the extraction of the PAs. The results were compared against heating under reflux. It was found that the binary mixture of MeOH:H₂O (1:1) acidified using HCl to pH 2-3 was the optimal solvent for the extraction of the PAs in the plant materials. Liquid chromatography (LC) with ultra-violet (UV) detection and electrospray ionization mass spectrometry (ESI-MS) in the positive mode was used for the determination and quantitation of senkirkine and senecionine in the botanical extract. The proposed extraction methods with LC/MS allow for the rapid detection of the major and the minor alkaloids in T. farfara in the presence of co-eluting peaks. With LC/MS, the quantitative analysis of PAs in the extract was done using internal standard calibration and the precision was found to vary from 0.6% to 5.4% on different days. The limits of detection (LODs) and limits of quantitation (LOQs) for MAE and PHWE were found to vary from 0.26 μg/g to 1.04 μg/g and 1.32 μg/g to 5.29 μg/g, respectively. The method precision of MAE and PHWE were found to vary from 3.7% to 10.4% on different days. The results showed that major and minor alkaloids extracted using MAE and PHWE were comparable to that by heating under reflux. Our data also showed that significant ion suppression was not observed in the analysis of senkirkine and senecionine in the botanical extracts with co-eluting peaks.     

Pressurized hot water extraction of bioactive or marker compounds in botanicals and medicinal plant materials

To reduce the use of organic solvent, pressurized hot water extraction (PHWE) has been shown to be a feasible option for the extraction of bioactive and marker compounds in botanicals and medicinal plants. The parameters that may affect the extraction efficiencies in PHWE include temperature, extraction time and addition of small percentage of organic solvent or surfactants. Currently, applications of PHWE for the extraction of thermally labile compounds in botanicals are still rather limited. PHWE with and without the additional of a small percentage of organic solvent such as ethanol is highly suited for the chemical standardization and quality control of medicinal plants. At the same time, it can be applied at the pilot scale as a manufacturing process for medicinal plants. Surfactant assisted PHWE was found to enhance the extraction of thermally labile and more hydrophobic species in medicinal plants at a lower temperature. The addition of small amount of surfactants in PHWE is highly suited for the determination of bioactive or marker compounds in medicinal plants. With proper optimization, PHWE was observed to have good extraction efficiency and precision when compared to other reference methods of extraction.     

Adsorption and photocatalytic degradation of methylene blue using high surface area titanate nanotubes (TNT) synthesized via hydrothermal method

Removal of methylene blue (MB) via adsorption and photocatalysis using titanate nanotubes (TNTs) with different surface areas were investigated and compared to commercial titanium dioxide (TiO₂) P25 Degussa nanoparticles. The TNTs with surface area ranging from 20 m²/g to 200 m²/g were synthesized via hydrothermal method with different reaction times. TEM imaging confirmed the tubular structure of TNT while XRD spectra indicated all TNTs exhibited anatase crystallinity. Batch adsorption rate showed linearity with surface properties of TNTs, where materials with higher surface area showed higher adsorption rate. The highest MB adsorption (70%) was achieved by TNT24 in 60 min whereas commercial TiO₂ exhibited the lowest adsorption of only 10% after 240 min. Adsorption isotherm studies indicated that adsorption using TNT is better fitted into Langmuir adsorption isotherm than Freundlich isotherm model. Furthermore, TNT24 was able to perform up to 90% removal of MB within 120 min, demonstrating performance that is 2-fold better compared to commercial TiO₂. The high surface area and surface Bronsted acidity are the main reasons for the improvement in MB removal performance exhibited by TNT24. The improvement in surface acidity enhanced the adsorption properties of all the nanotubes prepared in this study.     

Electrophoretic deposition of PVA coated hydroxyapatite on 316L stainless steel

Polyvinyl alcohol (PVA) coated hydroxyapatite was deposited onto a 316L stainless steel substrate by electrophoretic deposition. Deposition was carried out in a methanol suspension at pH 5.5 using a graphite rod as an anode. Parameters such as PVA concentration, deposition voltage and time were optimized to achieve a homogeneous, crack-free adhesive coating. Techniques such as X-ray diffraction and Fourier transform infrared spectroscopy were used to study the phase composition of the coated materials and the stability of hydroxyapatite in the presence of PVA.     

Precise measurement of the CP violation parameter sin2φ1 in B0→(cc¯)K0 decays

We present a precise measurement of the CP violation parameter sin2φ1 and the direct CP violation parameter A(f) using the final data sample of 772×10(6) BB[over ˉ] pairs collected at the Υ(4S) resonance with the Belle detector at the KEKB asymmetric-energy e(+)e(-) collider. One neutral B meson is reconstructed in a J/ψK(S)(0), ψ(2S)K(S)(0), χ(c1)K(S)(0), or J/ψK(L)(0) CP eigenstate and its flavor is identified from the decay products of the accompanying B meson. From the distribution of proper-time intervals between the two B decays, we obtain the following CP violation parameters: sin2φ1=0.667±0.023(stat)±0.012(syst) and A(f)=0.006±0.016(stat)±0.012(syst).