Design and synthesis of pyrrolo[3,2-d]pyrimidine HER2/EGFR dual inhibitors: Improvement of the physicochemical and pharmacokinetic profiles for potent in vivo anti-tumor efficacy

During the course of our studies on a novel HER2/EGFR dual inhibitor (TAK-285), we found an alternative potent pyrrolo[3,2-d]pyrimidine compound (1a). To enhance the pharmacokinetic (PK) profile of this compound, we conducted chemical modifications into its N-5 side chain and conversion of the chemically modified compounds into their salts. Among them, 2cb, the tosylate salt of compound 2c, showed potent HER2/EGFR kinase inhibitory activity (IC₅₀: 11/11 nM) and cellular growth inhibitory activity (BT-474 cell GI₅₀: 56 nM) with a good drug metabolism and PK (DMPK) profile. Furthermore, 2cb exhibited significant in vivo antitumor efficacy in both mouse and rat xenograft models with transplanted 4-1ST gastric cancer cell lines (mouse, T/C = 0%, 2cb po bid at 100 mg/kg; rat, T/C: -1%, 2cb po bid at 25 mg/kg).     

Biosynthesis of glycyrrhetic acid 3-O-mono-β-d-glucuronide catalyzed by β-d-glucuronidase with enhanced bond selectivity in an ionic liquid/buffer biphasic system

The bond selective hydrolysis of glycyrrhizin (GL) to glycyrrhetic acid 3-O-mono-β-d-glucuronide (GAMG) catalyzed by recombinant β-d-glucuronidase from Escherichia coli BL21 (PGUS-E) was successfully performed in an ionic liquid (IL)/buffer biphasic system. Five ILs were analyzed, however, a hydrophobic IL 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM]PF₆) showed the best biocompatibility with PGUS-E. An obvious enhancement in the initial reaction rate, substrate conversion, GAMG yield and chemical bond selectivity (S_cb) was observed using 40% (v/v) [BMIM]PF₆/buffer as the reaction medium when compared to the acetate buffer medium. Under the optimized conditions (pH 6.0, temperature 50 °C, substrate concentration 6 mM and shaking speed 200 rpm), the initial reaction rate, the GAMG yield and the S_cb reached 3.15 mM h⁻¹, 74.36% and 98.12%, respectively. The recyclability of [BMIM]PF₆ was also studied and found to be reusable for five batches with high recovery percentage (≥92%). Furthermore, the desired product and byproduct were easily separated since they were distributed in different phases. Additionally, higher V_max (3.14 versus 2.24 mM h⁻¹), lower apparent K_m (1.21 versus 1.80 mM) and E_a (25.97 versus 32.60 kJ mol⁻¹) were achieved in [BMIM]PF₆/buffer biphasic system than that in monophasic buffer system.     

Soil quality assessment of coastal wetlands in the Yellow River Delta of China based on the minimum data set

Little information is available to assess the dynamic changes in wetland soil quality in coastal regions, though it is essential for wetland conservation and management. Soil samples were collected in Suaeda salsa wetlands (SWs), Tamarix chinensis wetlands (TWs), Suaeda salsa–Tamarix chinensis wetlands (STWs), freshwater Phragmites australis wetlands (FPWs) and saltwater Phragmites australis wetlands (SPWs) in three sampling periods (i.e., summer and autumn of 2007 and spring of 2008). According to the flooding characteristics of these wetlands, the study area could be grouped into three sub-regions: short-term flooding region (STFR), seasonal flooding region (SFR) and tidal flooding region (TFR). Soil quality was evaluated using the soil quality index (SQI), which was calculated using the selected minimum data set (MDS) based on principal components analysis (PCA). Our results showed that soil salt content (SSC), total carbon (TC), magnesium (Mg), nitrate nitrogen (NO₃⁻-N) and total sulfur (TS) consisted of a MDS among 13 soil properties. The SQI values varied from 0.18 to 0.66 for all soil samples, of which the highest and lowest SQI values were observed in TFR. The average SQI values were significantly higher in summer (0.50 ± 0.13) than in spring (0.37 ± 0.13) and autumn (0.36 ± 0.11) in the whole study area (p < 0.05). The average SQI values followed the order STFR (0.44 ± 0.12) > TFR (0.41 ± 0.15) > SFR (0.35 ± 0.09) although no significant differences were observed among the three regions (p > 0.05). SPWs and SWs soils showed higher SQI values (0.50 ± 0.10 and 0.47 ± 0.15, respectively) than TWs (0.30 ± 0.08) soils (p < 0.05). The SSC was the dominant factor of soil quality with its proportion of 34.1% contributing to the SQI values, followed by TC (24.5%) and Mg (24.1%). Correlation analysis also showed that SQI values were significantly negatively correlated with SSC. SSC might be a characteristic indicator of wetland soil quality assessment in coastal regions. The findings of this study showed that the SQI based on MDS is a powerful tool for wetland soil quality assessment.     

Capacities of soil water reservoirs and their better regression models by combining “merged groups PCA” in Chongqing,China

Soil water resource, together with the surface and sub-surface water resource, is essential to the regional water balance and world water cycle. A total of 90 soil samples were collected from 30 different soil profiles of dry fields throughout Chongqing, China randomly to show how soil could be a crucial part of water resources by discussing their five types of calculated soil water reservoir capacities, namely the total soil water reservoir capacity (mm) (TC), soil water storage capacity (mm) (SC), unavailable soil water reservoir capacity (mm) (UC), available soil water reservoir capacity (mm) (AC), and soil dead water storage capacity (mm) (DC) in certain layer, respectively. Overall, the total soil water reservoir capacity in 0–40 cm was about 209 mm, of which 70 mm belonged to available soil water reservoir capacity. Not all the five types of soil water reservoir capacities had significant correlations between each other. Soil structure, especially the size and quantity of soil pore was mainly determined by soil particle composition (clay, silt, and sand content). The more sand and less clay led to the more soil macropores, which provided room for soil water. Thus, clay, silt, and sand content jointly produced profound influence on soil water reservoir capacities. Nevertheless, specific water capacity and topographic factors displayed weak correlations to soil water reservoir capacities, which required further research works. Ultimately, the better regression models were achieved by multiple regression analysis coupled with “merged groups PCA” than by multiple regression analysis with “all variables PCA”. UC, SC, TC and DC could be well simulated (mostly R² > 0.70; P < 0.05) through normal multiple regression analysis using original variables as well as multiple regression analysis with “merged groups PCA”. Only regression models of TC and DC were highly significant (mostly R² > 0.70; P < 0.05) through “all variables PCA” method. And there were poor coefficients of determination (R²) for AC (mostly R² < 0.40; P < 0.05) by all the three regression methods.     

Variation of soil nutrients and particle size under different vegetation types in the Yellow River Delta

The Yellow River Delta wetland, located at the southern coast of Bohai Gulf, provides important ecosystem services such as flood control, water purification, biodiversity conservation, nutrient removal and carbon sequestration, shoreline stabilization, tourism attraction and wetland products maintains in the Yellow River Delta. This study assessed how agricultural activities in a reclamation wetland changed soil pH, soil electric conductivity, soil nutrient and soil particle size as compared to natural vegetation by using a combination of field experiments and lab analysis. The vegetation type included adjacent alfalfa field (Medicago sativa), cotton field (Gossypium spp.), Chinese tamarisk shrub (Tamarix chinensis), and reed marsh (Phragmites sage). The results indicated that the soil pH was higher (pH > 8) in alfalfa field and cotton field, and alfalfa field and reed marsh had significant function in reducing soil salt content, soil electric conductivity of alfalfa field at 0–30 cm were 140.38 ± 14.36, 114.48 ± 14.36, 125.30 ± 11.37 μs/cm. The effect of different vegetation types on soil nutrient was significant (P < 0.05). Soil organic matter at 0–10 cm in Chinese tamarisk shrub and reed marsh was 21.66 ± 3.82 g/kg and 16.51 ± 4.60 g/kg, which was higher than that of alfalfa field (10.47 ± 2.36 g/kg) and cotton field (9.82 ± 1.27 g/kg), but soil total nitrogen content in alfalfa field was the highest, which is significantly higher than that of cotton field, Chinese tamarisk shrub and reed marsh(P < 0.05), the content of soil total nitrogen at 0–10 cm and 10–20 cm was 7.67 ± 0.38 g/kg and 5.97 ± 0.51 g/kg, respectively, while the content of available P and available K was reversed. The difference of soil particle size between layers was not significant (P > 0.05), the sand content of Chinese tamarisk shrub soils in 0–10 cm was the highest, the next was alfalfa field and cotton field, and the content of silt and clay in reed marsh was higher than the others. The correlation and significant degree between soil particle size and soil nutrient was related with vegetation types, and soil organic matter was significantly positively correlated with soil silt and clay content on the alfalfa field. The results demonstrated that wetland cultivation was one of the most important factors influencing on the nutrient fate and reserves in soil, which could lead to rapid nutrient release and slow restoration through abandon cultivation. Consequently, compared with cotton field, alfalfa field is more favorable to sustainable management of wetland cultivation in the Yellow River Delta. It should be considered in wetland restoration projects planning.     

Effect of phosphorylation on interaction of human tau protein with 14-3-3ζ

Interaction of the shortest isoform of tau protein (τ3) with human 14-3-3ζ was analyzed by means of native gel electrophoresis, chemical crosslinking and size-exclusion chromatography. Phosphorylation by cAMP-dependent protein kinase (up to 2 mole of phosphate per mole of τ3) strongly enhanced interaction of τ3 with 14-3-3. Apparent K_D of the complexes formed by phosphorylated τ3 and 14-3-3 was close to 2 μM, whereas the corresponding constant for unphosphorylated τ3 was at least 10 times higher. The stoichiometry of the complexes formed by phosphorylated τ3 and 14-3-3 was variable and was different from 1:1. 14-3-3 decreased the probability of formation of chemically crosslinked large homooligomers of phosphorylated τ3 and at the same time induced formation of crosslinked heterooligomeric complexes of τ3 and 14-3-3 with an apparent molecular mass of 120–140 kDa.     

Performance evaluation of various bioreactors for the removal of Cr(VI) and organic matter from industrial effluent

Performances of various bioreactors under different operating conditions were evaluated with respect to hexavalent chromium (Cr(VI)) reduction and COD removal. Continuous reactor studies were carried out with (i) aerobic suspended growth system, (ii) aerobic attached growth system, and (iii) anoxic attached growth system, using both synthetic and actual industrial wastewater. Arthrobacter rhombi-RE (MTCC7048), a Cr(VI) reducing strain enriched and isolated from chromium contaminated soil, was used in all the bioreactors for Cr(VI) biotransformation and COD removal. Aerobic and anoxic batch experiments were conducted to evaluate the bio-kinetic parameters. The bio-kinetic parameters for aerobic system were: μ_max = 2.34/d, K_s = 190 mg/L (as COD), K_i = 3.8 mg/L of Cr(VI), and Y_T = 0.377. These parameters for anoxic conditions were: μ_max = 0.57/d, K_s = 710 mg/L (as COD), K_i = 8.77 mg/L of Cr(VI), and Y_T = 0.13. Aerobic attached growth system, operated at a hydraulic retention time (HRT) of 24 h and an organic loading rate (OLR) of 3 kg/m³/d, performed better than aerobic suspended and the anoxic attached growth systems operated under identical conditions, while treating synthetic wastewater as well as industrial effluent.     

Comparative regional-scale soil salinity assessment with near-ground apparent electrical conductivity and remote sensing canopy reflectance

Soil salinity is recognized worldwide as a major threat to agriculture, particularly in arid and semi-arid regions. Producers and decision makers need updated and accurate maps of salinity in agronomically and environmentally relevant ranges (i.e., <20 dS m⁻¹, when salinity is measured as electrical conductivity of the saturation extract, EC_e). State-of-the-art approaches for creating accurate EC_e maps beyond field scale (i.e., 1 km²) include: (i) Analysis Of Covariance (ANOCOVA) of near-ground measurements of apparent soil electrical conductivity (EC_a) and (ii) regression modeling of multi-year remote sensing canopy reflectance and other co-variates (e.g., crop type, annual rainfall). This study presents a comparison of the two approaches to establish their viability and utility. The approaches were tested using 22 fields (total 542 ha) located in California’s western San Joaquin Valley. In 2013 EC_a-directed soil sampling resulted in the collection of 267 soil samples across the 22 fields, which were analyzed for EC_e, ranging from 0 to 38.6 dS m⁻¹. The ANOCOVA EC_a-EC_e model returned a coefficient of determination (R²) of 0.87 and root mean square prediction error (RMSPE) of 3.05 dS m⁻¹. For the remote sensing approach seven years (2007–2013) of Landsat 7 reflectance were considered. The remote sensing salinity model had R² = 0.73 and RMSPE = 3.63 dS m⁻¹. The robustness of the models was tested with a leave-one-field-out (lofo) cross-validation to assure maximum independence between training and validation datasets. For the ANOCOVA model, lofo cross-validation provided a range of scenarios in terms of RMSPE. The worst, median, and best fit scenarios provided global cross-validation R² of 0.52, 0.80, and 0.81, respectively. The lofo cross-validation for the remote sensing approach returned a R² of 0.65. The ANOCOVA approach performs particularly well at EC_e values <10 dS m⁻¹, but requires extensive field work. Field work is reduced considerably with the remote sensing approach, but due to the larger errors at low EC_e values, the methodology is less suitable for crop selection, and other practices that require accurate knowledge of salinity variation within a field, making it more useful for assessing trends in salinity across a regional scale. The two models proved to be viable solutions at large spatial scales, with the ANOCOVA approach more appropriate for multiple-field to landscape scales (1–10 km²) and the remote sensing approach best for landscape to regional scales (>10 km²).     

Assessment of wind and water erosion risk in the watershed of the Ningxia-Inner Mongolia Reach of the Yellow River,China

The watershed of the Ningxia-Inner Mongolia Reach of the Yellow River (NIMRYR) suffers from soil erosion by wind and water because of the wide distribution of deserts and arsenic sandstones in this region. The sediment generated by erosion fed into the Yellow River directly or by its tributaries, silting up the Yellow River and raising the elevation of the river bed. The silting of the Yellow River result in serious flood disasters in this watershed. Therefore, it is urgent to implement soil conservation projects to control wind and water soil erosion. To reach this objective, understanding the spatial and temporal variations of soil erosion in this watershed is very important. In this study, an assessment of soil erosion risk by wind and water was performed based on soil erosion models. The Integrated Wind Erosion Modeling System (IWEMS) and the Revised Wind Erosion Equation (RWEQ) were used to estimate the wind erosion modulus in this watershed, and the water erosion modulus was estimated by the Revised Universal Soil Loss Equation (RUSLE). The results show that during 2000s, the wind erosion modulus ranged from 0 to 31,440.4 t/km²/a, and the water erosion modulus was from 0 to 24,048.5 t/km²/a. Moreover, the total soil erosion modulus by wind and water has ranged from 0 to 32,792.7 t/km²/a. Due to the influence of regional weather and geomorphology, occurrences of wind and water erosion in this watershed are not identical in their spatial and temporal patterns. Based on the calculated soil erosion modulus, soil erosion risk was assessed according to the “Classification criteria for soil-erosion intensities” (SL190-2007). It was assumed that the areas with erosion intensity higher than slight were at risk of erosion; by this criterion, more than 34% of the total area of the watershed of the NIMRYR would be at erosion risk. Based on this estimation, it was also found that the NIMRYR watershed is not a region of wind–water erosion crisscross and that land-use conversions have a significant impact on soil erosion.     

Soil salinity and drought alter wood density and vulnerability to xylem cavitation of baldcypress (Taxodium distichum (L.) Rich.) seedlings

We investigated the role of hydraulic conductivity, wood density, and xylem cavitation in the response of baldcypress (Taxodium distichum) seedlings to increased soil salinity and drought. One-year-old, greenhouse-grown seedlings were irrigated daily with a 100 mM (≈6‰) salt solution or once per week with fresh water (drought). Controls were irrigated daily with fresh water. Gas exchange rates of stressed plants were reduced by approximately 50% (salt) and 70% (drought), resulting in a 50–60% reduction in diameter growth for both treatments. Stem-specific hydraulic conductivity (K_{S native}) of stressed plants was 33% (salt) and 66% (drought) lower than controls and we observed a strong positive correlation between K_{S native} and gas exchange. In addition, we found a strong relationship between CO₂ assimilation rate (A) and the soil-to-leaf hydraulic conductance (k_L). The relationship was identical for all treatments, suggesting that our moderate salt stress (as well as drought) did not affect the photosynthetic biochemistry of leaves, but rather reduced A via stomatal closure. Lower K_{S native} of stressed plants was associated with increased wood density and greater resistance to xylem cavitation. Xylem pressures causing 50% loss of hydraulic conductivity (P₅₀) were ?2.88 ± 0.07 MPa (drought), ?2.50 ± 0.08 MPa (salt) and ?2.01 ± 0.04 MPa (controls). P₅₀s were strongly correlated with wood density (r = ?0.71, P < 0.01) and K_{S native} (r = 0.74, P < 0.01). These findings support the hypothesis that there is a significant trade-off between a plant's cavitation resistance and its hydraulic efficiency. The results of the present study indicate that stressed plants partitioned their biomass in a way that strengthened their xylem and reduced vulnerability to xylem cavitation. Hence, these seedlings could be better suited to be planted in environments with elevated soil salinity. For most parameters (especially P₅₀), drought had an even more pronounced effect than salinity. This is important as nurseries could produce “stress-acclimated” seedlings simply by reducing irrigation amounts and would not have to contaminate the soils in their nursery beds with salt applications.     

Synthesis and monoamine transporter affinity of 3α-arylmethoxy-3β-arylnortropanes

A series of 3-arylnortrop-2-enes and 3α-arylmethoxy-3β-arylnortropanes were synthesized and evaluated for binding affinity at monoamine transporters. The 3-(3,4-dichlorophenyl)nortrop-2-ene (6e) exhibited high affinity for the SERT (K_i = 0.3 nM). The 3α-arylmethoxy-3β-arylnortropanes were generally SERT selective with the 3α-(3.4-dichlorophenylmethoxy)-3βphenylnortrop-2-ene (7c) possessing subnanomolar potency (K_i = 0.061 nM). However, 3α-(3,4-dichlorophenylmethoxy)-3β-phenylnortrop-2-ene (7b) exhibited high affinity at all three transporters [(DAT K_i = 22 nM), (SERT K_i = 6 nM) and (NET K_i = 101 nM)].     

Pseudo-enzymatic hydrolysis of 4-nitrophenyl acetate by human serum albumin: pH-dependence of rates of individual steps

Human serum albumin (HSA) displays esterase activity reflecting multiple irreversible chemical modifications rather than turnover. Here, kinetics of the pseudo-enzymatic hydrolysis of 4-nitrophenyl acetate (NphOAc) are reported. Under conditions where [HSA] ? 5×[NphOAc] and [NphOAc] ? 5×[HSA], the HSA-catalyzed hydrolysis of NphOAc is a first-order process for more than 95% of its course. From the dependence of the apparent rate constants k_app and k_obs on [HSA] and [NphOAc], respectively, values of K_s, k₊₂, and k₊₂/K_s were determined. Values of K_s, k₊₂, and k₊₂/K_s obtained at [HSA] ? 5×[NphOAc] and [NphOAc] ? 5×[HSA] are in good agreement, the deacylation step being rate limiting in catalysis. The pH-dependence of k₊₂/K_s, k₊₂, and K_s reflects the acidic pK_a shift of the Tyr411 catalytic residue from 9.0 ± 0.1 in the substrate-free HSA to 8.1 ± 0.1 in the HSA:NphOAc complex. Accordingly, diazepam inhibits competitively the HSA-catalyzed hydrolysis of NphOAc by binding to Tyr411.     

Inhibition kinetics of a commercial mixed culture by ammonium sulfate

The inhibitory effect of ammonium sulfate on a commercial mixed culture, used in biological waste-water treatment was studied under aerobic batch conditions. Several mathematical models of enzyme and growth kinetics including a death factor were analyzed through nonlinear regression to find the best fit to corresponding data of inhibition. The best fit model was found to be the generalized Monod type with a death factor having the biokinetic parameters; μ_max 0.681 h⁻¹, K_s 0.224 g dm⁻³, K_i 56240 g dm⁻³, K 0.055 g dm⁻³ and k_d 0.052 h⁻¹ to represent the experimental data accurately. The low saturation coefficient value along with high maximum specific growth rate and inhibition coefficient denotes the competitive characteristics of commercial mixed cultures in the biological treatment of high ammonium polluted waste waters.     

Effects of grassland conversion to cropland and forest on soil organic carbon and dissolved organic carbon in the farming-pastoral ecotone of Inner Mongolia

Effects of grassland conversion to cropland and forest on soil organic carbon (SOC), dissolved organic carbon (DOC) in the farming-pastoral ecotone of Inner Mongolia were investigated by direct field sampling. SOC content and DOC content in soil decreased after grassland were shifted to forest or cropland, in the sequence of grassland soil > forest soil > cropland soil. SOC stock declined by 18% after grassland shifted from to forest. Reclamation of cropland for 10 years, 15 years and 20 years lost SOC in 0–30 cm soil layer, by 34%, 14% and 18%, respectively, compared with that of grassland. DOC in 3 soil layers was within 21.1–26.5 mg/L in grassland, 12.1–14.6 mg/L in forest soil, and 8.0–14.0 mg/L in cropland soil. Correlation analysis indicated that SOC content and DOC content were positively dependent on total nitrogen content (p < 0.05), but negatively on bulk density or land use type (p < 0.05). DOC was positively correlated SOC (p < 0.01). Moreover, SOC content could be quantitatively described by a linear combination of land use types (p = 0.000, r² = 0.712), and DOC content by a linear combination of two soil-related variables, land use types and SOC (p = 0.000, r² = 0.861).     

9-Aminomethyl-9,10-dihydroanthracene (AMDA) analogs as structural probes for steric tolerance in 5-HT2A and H1 receptor binding sites

Synthesis, radioligand binding and molecular modeling studies of several 9-aminomethyl-9,10-dihydroanthracene (AMDA) analogs were carried out to determine the extent of the steric tolerance associated with expansion of the tricyclic ring system and amine substitution at 5-HT_2A and H₁ receptors. A mixture of (7,12-dihydrotetraphene-12-yl)methanamine and (6,11-dihydrotetracene-11-yl)methanamine in a 75–25% ratio was found to have an apparent K_i of 10 nM at the 5-HT_2A receptor. A substantial binding affinity for (7,12-dihydrotetraphene-3-methoxy-12-yl)methanamine at the 5-HT_2A receptor (K_i = 21 nM) was also observed. Interestingly, this compound was found to have 100-fold selectivity for 5-HT_2A over the H₁ receptor (K_i = 2500 nM). N-Phenylalkyl-AMDA derivatives, in which the length of the alkyl chain varied from methylene to n-butylene, were found to have only weak affinity for both 5-HT_2A and H₁ receptors (K_i = 223 to 964 nM). Our results show that large rigid annulated AMDA analogs can be sterically accommodated within the proposed 5-HT_2A binding site.     

Effect of stand age on soil microbial community structure in wolfberry (Lycium barbarum L.) fields

Soil physicochemical properties and microbes are essential in terrestrial ecosystems through their role in cycling mineral compounds and decomposing organic matter. This study examined the effect of stand age on soil physicochemical properties and microbial community structure in wolfberry (Lycium barbarum L.) fields, in order to reveal the mechanism of soil degradation due to long-term stand of L. barbarum. The objective of the study was achieved by phospholipid fatty acid (PLFA) biomarker analysis of soil samples from L. barbarum fields in Zhongning County, Ningxia Province—the origin of L. barbarum. Five stand ages of L. barbarum were selected, < 1, 3, 6, 9, and 12 years (three plots each). The results showed that soil bulk density increased slightly with increasing stand age, while no clear trend was observed in soil pH or total salinity. As the stand age increased, soil organic matter and nutrients first increased before decreasing, with the highest levels being found in year 9. There was an amazing variety of PLFA biomarkers in soil samples at different stand ages. The average concentrations of total, bacterial, fungal, and actinomycete PLFAs in the surface soil initially decreased and then increased, before decreasing with the stand age in summer. The PLFA concentrations of major microbial groups were highest in year 9, with the total PLFA concentrations being 32.97% and 10.67% higher than those in years < 1 and 12, respectively. Higher microbial PLFA concentrations were detected in summer relative to autumn and in the surface relative to the subsurface soil. The highest ratios of Gram-positive to Gram-negative bacterial (G^?/G⁺) and fungal to bacterial (F/B) PLFAs were obtained in year 6, on average 76.09% higher than those at the other four stand ages. The soil environment was most stable in year 6, with no differences between other stand ages. Therefore, soil microbial community structure was strongly influenced by the stand age in year 6 only. The effect of stand age on soil G^?/G⁺ and microbial community structure varied with season and depth; there was little effect for F/B in the 20–40 cm soil layer. Principal component analysis revealed no correlations between microbial PLFA concentrations and total salinity in the soil; negative correlations were noted between soil pH and F/B in summer (P < 0.01), as well as between soil pH and fungal PLFA in autumn (P < 0.05). Moreover, microbial PLFA concentrations were correlated with soil organic matter (mean R = 0.7725), total nitrogen (mean R = 0.8296), total phosphorus (mean R = 0.8175), available nitrogen (mean R = 0.7458), and available phosphorus (mean R = 0.7795) (P < 0.01). On the whole, the soil ecosystem was most stable in year 6, while soil organic matter, nutrients, and microbial PLFA concentrations were maximal in year 9; thereafter, soil fertility indices and microbial concentrations decreased and soil quality declined gradually as the stand age increased. Therefore, farmers should reduce the application rate of fertilizers, especially compound or mixed fertilizers, in L. barbarum fields; organic or bacterial manure can be applied increasingly to improve the soil environment and prolong the economic life of L. barbarum.     

Improved binding affinities of pyrrolidine derivatives as Mcl-1 inhibitors by modifying amino acid side chains

As an important member of anti-apoptotic Bcl-2 protein, myeloid cell leukemia sequence 1 (Mcl-1) protein is an attractive target for cancer therapy. In this study, a new series of pyrrolidine derivatives as Mcl-1 inhibitors were developed by mainly modifying the amino acid side chain of compound 1. Among them, compound 18 (K_i = 0.077 μM) exhibited better potent inhibitory activities towards Mcl-1 protein compared to positive control Gossypol (K_i = 0.18 μM). In addition, compound 40 possessed good antiproliferative activities against PC-3 cells (K_i = 8.45 μM), which was the same as positive control Gossypol (K_i = 7.54 μM).     

Asymmetric total synthesis and identification of tetrahydroprotoberberine derivatives as new antipsychotic agents possessing a dopamine D1, D2 and serotonin 5-HT1A multi-action profile

An effective and rapid method for the microwave-assisted preparation of the key intermediate for the total synthesis of tetrahydroprotoberberines (THPBs) including l-stepholidine (l-SPD) was developed. Thirty-one THPB derivatives with diverse substituents on A and D ring were synthesized, and their binding affinity to dopamine D₁, D₂ and serotonin 5-HT_1A and 5-HT_2A receptors were determined. Compounds 18k and 18m were identified as partial agonists at the D₁ receptor with K_i values of 50 and 6.3 nM, while both compounds act as D₂ receptor antagonists (K_i = 305 and 145 nM, respectively) and 5-HT_1A receptor full agonists (K_i = 149 and 908 nM, respectively). These two THPBs compounds exerted antipsychotic actions in animal models. Further electrophysiological studies employing single-unit recording in intact animals demonstrated that 18k-excited dopaminergic (DA) neurons are associated with its 5-HT_1A receptor agonistic activity. These results suggest that these two compounds targeted to multiple neurotransmitter receptors may present novel lead drugs with new pharmacological profiles for the treatment of schizophrenia.     

NAD-based inhibitors with anticancer potential

Three classes of novel inhibitors of inosine monophosphate dehydrogenase have been prepared and their anti-proliferative properties were evaluated against several cancer cell lines.(1) Mycophenolic adenine dinucleotide analogues (8–13) containing a substituent at the C2 of adenine ring were found to be potent inhibitors of IMPDH (K_i’s in range of 0.6–82 nM) and sub-μM inhibitors of leukemic K562 cell proliferation. (2) Mycophenolic adenosine (d and l) esters (20 and 21) showed a potent inhibition of IMPDH2 (K_i = 102 and K_i = 231 nM, respectively) and inhibition of K562 cell growth (IC₅₀ = 0.5 and IC₅₀ = 1.6 μM). These compounds serve both as inhibitors of the enzyme and as a depot form of mycophenolic acid. The corresponding amide analogue 22, also a potent inhibitor of IMPDH (K_i = 84 nM), did not inhibit cancer cell proliferation. (3) Mycophenolic-(l)- and (d)-valine adenine di-amide derivatives 25 (K_i = 9 nM) and 28 (K_i = 3 nM) were found to be very potent enzymatically, but did not inhibit proliferation of cancer cells.     

Discovery of novel α1-adrenoceptor ligands based on the antipsychotic sertindole suitable for labeling as PET ligands

The synthesis and in vitro preclinical profile of a series of 5-heteroaryl substituted analogs of the antipsychotic drug sertindole are presented. Compounds 1-(4-fluorophenyl)-3-(1-methylpiperidin-4-yl)-5-(pyrimidin-5-yl)-1H-indole (Lu AA27122, 3i) and 1-(4-fluorophenyl)-5-(1-methyl-1H-1,2,4-triazol-3-yl)-3-(1-methylpiperidin-4-yl)-1H-indole (3l) were identified as high affinity α_1A-adrenoceptor ligands with K_i values of 0.52 and 0.16 nM, respectively, and with a >100-fold selectivity versus dopamine D₂ receptors. Compound 3i showed almost equal affinity for α_1B- (K_i = 1.9 nM) and α_1D-adrenoceptors (K_i = 2.5 nM) as for α_1A, as well as moderate affinity for 5-HT_1B (K_i = 13 nM) and 5-HT₆ (K_i = 16 nM) receptors, whereas 3l showed >40-fold selectivity toward all other targets tested. Based on in vitro assays for assessment of permeability rates and extent, it is predicted that both compounds enter the brain of rats, non-human primates, as well as humans, and as such are good candidates to be carried forward for further evaluation as positron emission tomography (PET) ligands.