Green approach to the synthesis of α-aminophosphonate-tetrahydroisoquinoline hybrids and their anti-cholinesterase activity.

A series of 19 novel α-aminophosphonate-tetrahydroisoquinoline hybrids were synthesized through a cross dehydrogenative coupling reaction between N-aryl-tetrahydroisoquinolines and dialkylphosphites, using tert-butyl hydroperoxide as oxidazing agent. This simple procedure provided products with high atom economy and moderate to high yields. In vitro cholinesterase inhibitory activity of these compounds was evaluated. All the synthesized compounds showed good to excellent selective inhibition against butyrylcholinesterase. Compound 3bc was found to be the most active derivative with an IC50 of 9 nM. Molecular modelling studies suggested that the inhibitor is located in the peripheral anionic site (PAS) of the enzyme and interacts with some residue of the catalytic anionic site. Kinetic studies revealed that 3bc acts as a non-competitive inhibitor. Predicted ADME showed good pharmacokinetics and drug-likeness properties for most hybrids. Each newly synthesized compound was characterized by IR, 1H NMR, 13C NMR, 31P NMR spectral studies and also HRMS. The results of this study suggest that α-aminophosphonate-tetrahydroisoquinoline hybrids can be promising lead compounds in the discovery of new and improved drugs for the treatment of Alzheimer's disease and related neurodegenerative disorders.

Putting the Puzzle Together To Get the Whole Picture: Molecular Basis of the Affinity of Two Steroid Derivatives to Acetylcholinesterase.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder that has no cure because its etiology is still unknown, and its main treatment is the administration of acetylcholinesterase (AChE) inhibitors. The study of the mechanism of action of this family of compounds is critical for the design of new more potent and specific inhibitors. In this work, we study the molecular basis of an uncompetitive inhibitor (compound 1, 2ß, 3α-dihydroxy-5α-cholestan-6-one disulfate), which we have proved to be a peripheral anionic site (PAS)-binding AChE inhibitor. The pipeline designed in this work is key to the development of other PAS inhibitors that not only inhibit the esterase action of the enzyme but could also modulate the non-cholinergic functions of AChE linked to the process of amylogenesis. Our studies showed that 1 inhibits the enzyme not simply by blocking the main gate but by an allosteric mechanism. A detailed and careful analysis of the ligand binding position and the protein dynamics, particularly regarding their secondary gates and active site, was necessary to conclude this. The same analysis was executed with an inactive analogue (compound 2, 2ß, 3α-dihydroxy-5α-cholestan-6-one). Our first computational results showed no differences in affinity to AChE between both steroids, making further analysis necessary. This work highlights the variables to be considered and develops a refined methodology, for the successful design of new potent dual-action drugs for AD, particularly PAS inhibitors, an attractive strategy to combat AD.

Design and Microwave-Assisted Synthesis of Aza-Resveratrol Analogs with Potent Cholinesterase Inhibition.

BACKGROUND: Currently approved Alzheimer's disease medications mainly comprise acetylcholinesterase inhibitors. Many of these inhibitors are either natural compounds or synthetic molecules inspired in natural compounds. Hybrid molecules that can interact with different target sites of the enzyme could lead to the discovery of effective multitarget drugs. OBJECTIVE: To design, synthesize, and evaluate a series of new aza-resveratrol analogs as in vitro acetyl- and butyrylcholinesterase inhibitors. METHODS: The synthesis is achieved by a simple and efficient microwave-assisted method, from commercially available starting materials. Compounds are designed as hybrids of an aza-stilbene nucleus (Schiff base) connected to a tertiary amine by a hydrocarbon chain of variable length, designed to interact both with the peripheric anionic site and the catalytic site of the enzyme. RESULTS: All the derivatives inhibit both enzymes in a concentration-dependent manner, acting as moderate to potent cholinesterase inhibitors. The most potent inhibitors are compounds 12b (IC50 = 0.43 µM) and 12a (IC50 = 0.31 µM) for acetyl- and butyrylcholinesterase, respectively. Compounds 12a and 12b also exhibit significant acetylcholinesterase inhibition in SH-SY5Y human neuroblastoma cells without cytotoxic properties. Enzyme kinetic studies and molecular modeling reveal that inhibitor 12b targets both the catalytic active site and the peripheral anionic site of acetylcholinesterase what makes it able to modulate the self-induced ß-amyloid aggregation. Furthermore, the molecular modeling analysis helps to assess the impact of the linker length in the inhibitory activity of this family of new cholinesterase inhibitors. CONCLUSION: These compounds have the potential to serve as a dual binding site inhibitor and might provide a useful template for the development of new anti-Alzheimer's disease agents.

Noncontact Imaging of Ion Dynamics in Polymer Electrolytes with Time-Resolved Electrostatic Force Microscopy.

Ionic-transport processes govern performance in many classic and emerging devices, ranging from battery storage to modern mixed-conduction organic electrochemical transistors (OECT). Here, we study local ion-transport dynamics in polymer films using time-resolved electrostatic force microscopy (trEFM). We establish a correspondence between local and macroscopic measurements using local trEFM and macroscopic electrical impedance spectroscopy (EIS). We use polymer films doped with lithium bis(trifluoromethane)sulfonimide (LiTFSI) as a model system where the polymer backbone has oxanorbornenedicarboximide repeat units with an oligomeric ethylene oxide side chain of length n. Our results show that the local polymer response measured in the time domain with trEFM follows stretched-exponential relaxation kinetics, consistent with the Havriliak-Negami relaxation we measure in the frequency-domain EIS data for macroscopic samples of the same polymers. Furthermore, we show that the trEFM results capture the same trends as the EIS results-changes in ion dynamics with increasing temperature, increasing salt concentration, and increasing volume fraction of ethylene oxide side chains in the polymer matrix evolve with the same trends in both measurement techniques. We conclude from this correlation that trEFM data reflect, at the nanoscale, the same ionic processes probed in conventional EIS at the device level. Finally, as an example application for emerging materials syntheses, we use trEFM and infrared photoinduced force microscopy (PiFM) to image a diblock copolymer electrolyte for next-generation solid-state energy storage applications.

A Minimalist Approach to the Design of Complexity-Enriched Bioactive Small Molecules: Discovery of Phenanthrenoid Mimics as Antiproliferative Agents.

Over the last decades, much effort has been devoted to the design of the "ideal" library for screening, the most promising strategies being those which draw inspiration from biogenic compounds, as the aim is to add biological relevance to such libraries. On the other hand, there is a growing understanding of the role that molecular complexity plays in the discovery of new bioactive small molecules. Nevertheless, the introduction of molecular complexity must be balanced with synthetic accessibility. In this work, we show that both concepts can be efficiently merged-in a minimalist way-by using very simple guidelines during the design process along with the application of multicomponent reactions as key steps in the synthetic process. Natural phenanthrenoids, a class of plant aromatic metabolites, served as inspiration for the synthesis of a library in which complexity-enhancing features were introduced in few steps using multicomponent reactions. These resulting chemical entities were not only more complex than the parent natural products, but also interrogated an alternative region of the chemical space, which led to an outstanding hit rate in an antiproliferative assay: four out of twenty-six compounds showed in vitro activity, one of them being more potent than the clinically useful drug 5-fluorouracil.

Oxidation at C-16 enhances butyrylcholinesterase inhibition in lupane triterpenoids.

A set of triterpenoids with different grades of oxidation in the lupane skeleton were prepared and evaluated as cholinesterase inhibitors. Allylic oxidation with selenium oxide and Jones's oxidation were employed to obtain mono-, di- and tri-oxolupanes, starting from calenduladiol (1) and lupeol (3). All the derivatives showed a selective inhibition of butyrylcholinesterase over acetylcholinesterase (BChE vs. AChE). A kinetic study proved that compounds 2 and 9, the more potent inhibitors of the series, act as competitive inhibitors. Molecular modeling was used to understand their interaction with BChE, the role of carbonyl at C-16 and the selectivity towards this enzyme over AChE. These results indicate that oxidation at C-16 of the lupane skeleton is a key transformation in order to improve the cholinesterase inhibition of these compounds.

Impact and implementation of a sustainable regional telestroke network.

BACKGROUND: Telestroke uses videoconferencing technology to allow off-site experts to provide stroke thrombolysis decision support to less experienced front line clinicians. AIM: To assess the impact of a new telestroke service on thrombolysis rates and door-to-needle times in participating provincial hospitals and service resources to aid transition to a sustainable telestroke service. METHODS: This is a sequential comparison of 'pre' (December 2015 to May 2016) and 'post' (June 2016 to December 2016) implementation outcomes. The main outcomes were thrombolysis rate and door-to-needle time. All patient data were captured prospectively in a central database. Data captured and analysed also included technical problems, consumer and clinician feedback, and additional service resources required. RESULTS: Over the study period, 164 telestroke assessments were completed, including the 'hub' hospital. Among the participating provincial hospitals, 21 of 343 patients (6.1%) were thrombolysed in the 6-months prior to June 2016 and 50 of 318 patients (15.7%) during the 6-month following implementation of telestroke; odds ratio 2.86 (95% confidence interval 1.68-4.89); P = 0.0001. Overall, mean (standard deviation) regional hospital door-to-needle time reduced from 79.6 (31.4) to 62.7 (23.3) min (P = 0.015). Videoconferencing failure occurred in 4.8% of cases. Consumer and clinician feedback was positive. The main resource challenge was doubling of out-of-hours neurologist workload. CONCLUSION: Telestroke was associated with a significant increase in thrombolysis rate and reduction in door-to-needle time in provincial hospitals indicating improved patient care. Quantification of the extra neurologist workload allowed for a seamless transition to 'business as usual' using a novel annual subscription funding and service model.

Polyhydroxylated sulfated steroids derived from 5α-cholestanes as antiviral agents against herpes simplex virus.

Twelve polyhydroxylated sulfated steroids synthesized from a 5α-cholestane skeleton with different substitutions in C-2, C-3 and C-6 were evaluated for cytotoxicity and antiviral activity against herpes simplex virus (HSV) by a virus plaque reduction assay. Four compounds elicited a selective inhibitory effect against HSV. The disodium salt of 2ß,3α-dihydroxy-6E-hydroximine-5α-cholestane-2,3-disulfate, named compound 7, was the most effective inhibitor of HSV-1, HSV-2 and pseudorabies virus (PrV) strains, including acyclovir-resistant variants, in human and monkey cell lines. Preliminary mechanistic studies demonstrated that compound 7 did not affect the initial steps of virus entry but inhibited a subsequent event in the infection process of HSV.

Prediction of Core Body Temperature from Multiple Variables.

This paper aims to improve the prediction of rectal temperature (T re) from insulated skin temperature (T is) and micro-climate temperature (T mc) previously reported (Richmond et al., Insulated skin temperature as a measure of core body temperature for individuals wearing CBRN protective clothing. Physiol Meas 2013; 34:1531-43.) using additional physiological and/or environmental variables, under several clothing and climatic conditions. Twelve male (25.8±5.1 years; 73.6±11.5kg; 178±6cm) and nine female (24.2±5.1 years; 62.4±11.5kg; 169±3cm) volunteers completed six trials, each consisting of two 40-min periods of treadmill walking separated by a 20-min rest, wearing permeable or impermeable clothing, under neutral (25°C, 50%), moderate (35°C, 35%), and hot (40°C, 25%) conditions, with and without solar radiation (600W m(-2)). Participants were measured for heart rate (HR) (Polar, Finland), skin temperature (T s) at 11 sites, T is (Grant, Cambridge, UK), and breathing rate (f) (Hidalgo, Cambridge, UK). T mc and relative humidity were measured within the clothing. T re was monitored as the 'gold standard' measure of T c for industrial or military applications using a 10cm flexible probe (Grant, Cambridge, UK). A stepwise multiple regression analysis was run to determine which of 30 variables (T is, T s at 11 sites, HR, f, T mc, temperature, and humidity inside the clothing front and back, body mass, age, body fat, sex, clothing, Thermal comfort, sensation and perception, and sweat rate) were the strongest on which to base the model. Using a bootstrap methodology to develop the equation, the best model in terms of practicality and validity included T is, T mc, HR, and 'work' (0 = rest; 1 = exercise), predicting T re with a standard error of the estimate of 0.27°C and adjusted r (2) of 0.86. The sensitivity and specificity for predicting individuals who reached 39°C was 97 and 85%, respectively. Insulated skin temperature was the most important individual parameter for the prediction of T re. This paper provides novel information about the viability of predicting T c under a wide range of conditions, using predictors which can practically be measured in a field environment.

Small Molecules as Anti-TNF Drugs.

Tumor necrosis factor (TNF, TNF-α, cachectin) is a pleiotropic, proinflammatory cytokine with multiple biological effects, many of which are not yet fully understood. Although TNF was initially described as an anti-tumor agent more than three decades ago, current knowledge places it central to immune system homeostasis. TNF plays a critical role in host defense against infection, as well as an inhibitory role in autoimmune disease. However, TNF overproduction generates deleterious effects by inducing the transcription of genes involved in acute and chronic inflammatory responses including asthma, rheumatoid arthritis, Crohn's disease, and psoriasis. Direct inhibition of TNF by biologics, such as monoclonal antibodies and circulating TNF receptor constructs, has produced effective treatments for these disorders and validated the inhibition of this proinflammatory cytokine as an effective therapy. Unfortunately, these biological therapies suffer from several drawbacks, including high cost and the induction of autoantibody production. Thus, the development of small molecules able to modulate TNF production or signaling pathways remains a central challenge in Medicinal Chemistry. Considerable efforts have been made over the past two decades to develop such inhibitors, which could potentially be administered orally and would presumably be cheaper. This review is focused on the recent development of compounds that modulate the activity of this cytokine by acting at different levels, such as TNF expression, processing, binding to its receptors and direct inhibition. These approaches will be compared and discussed.

Synthesis and cholinesterase inhibition of cativic acid derivatives.

Alzheimer's disease (AD) is a neurodegenerative disorder associated with memory impairment and cognitive deficit. Most of the drugs currently available for the treatment of AD are acetylcholinesterase (AChE) inhibitors. In a preliminary study, significant AChE inhibition was observed for the ethanolic extract of Grindelia ventanensis (IC50=0.79 mg/mL). This result prompted us to isolate the active constituent, a normal labdane diterpenoid identified as 17-hydroxycativic acid (1), through a bioassay guided fractionation. Taking into account that 1 showed moderate inhibition of AChE (IC50=21.1 µM), selectivity over butyrylcholinesterase (BChE) (IC50=171.1 µM) and that it was easily obtained from the plant extract in a very good yield (0.15% w/w), we decided to prepare semisynthetic derivatives of this natural diterpenoid through simple structural modifications. A set of twenty new cativic acid derivatives (3-6) was prepared from 1 through transformations on the carboxylic group at C-15, introducing a C2-C6 linker and a tertiary amine group. They were tested for their inhibitory activity against AChE and BChE and some structure-activity relationships were outlined. The most active derivative was compound 3c, with an IC50 value of 3.2 µM for AChE. Enzyme kinetic studies and docking modeling revealed that this inhibitor targeted both the catalytic active site and the peripheral anionic site of this enzyme. Furthermore, 3c showed significant inhibition of AChE activity in SH-SY5Y human neuroblastoma cells, and was non-cytotoxic.

Preparation, anticholinesterase activity and molecular docking of new lupane derivatives.

A set of twenty one lupane derivatives (2-22) was prepared from the natural triterpenoid calenduladiol (1) by transformations on the hydroxyl groups at C-3 and C-16, and also on the isopropenyl moiety. The derivatives were tested for their inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) and some structure-activity relationships were outlined with the aid of enzyme kinetic studies and docking modelization. The most active compound resulted to be 3,16,30-trioxolup-20(29)-ene (22), with an IC50 value of 21.5µM for butyrylcholinesterase, which revealed a selective inhibitor profile towards this enzyme.

Energy balance and physical demands during an 8-week arduous military training course.

This study assessed soldier's physical demands and energy balance during the Section Commanders' Battles Course (SCBC). Forty male soldiers were monitored during the 8-week tactics phase of the SCBC. Energy expenditure was measured using the doubly labeled water method. Cardiovascular strain (heart rate) and physical activity (using triaxial accelerometer) were also monitored. Average sized portions of meals were weighed, with all recipes and meals entered into a dietary analysis program to calculate the calorie content. Energy expenditure averaged 19.6 ± 1.8 MJ · d(-1) in weeks 2 to 3 and 21.3 ± 2.0 MJ · d(-1) in weeks 6 to 7. Soldiers lost 5.1 ± 2.6 kg body mass and body fat percent decreased from 23 ± 4% to 19 ± 5%. This average weight loss equates to an estimated energy deficit of 2.69 MJ · d(-1). The Army provided an estimated 14.0 ± 2.2 MJ · d(-1) in weeks 2 to 3 and 15.7 ± 2.2 MJ · d(-1) in weeks 6 to 7. Although this provision adheres to the minimum requirement of 13.8 MJ · d(-1) set by Army regulations, soldiers were in a theoretical 5.6 MJ · d(-1) energy deficit. The physical demands of SCBC were high, and soldiers were in energy deficit resulting in loss in body mass; primarily attributed to a loss in fat mass.

Synthesis and cytotoxic evaluation of four new 6E-hydroximinosteroids.

Four new 6E-hydroximinosteroids (1, 2a, 3 and 4) have been synthesized from the corresponding ketones, 2ß,3ß-dihydroxy-5α-cholestan-6-one (5), 2α,3α-dihydroxy-5α-cholestan-6-one (6), 2ß,3α-dihydroxy-5α-cholestan-6-one (7) and 2ß,3α-dihydroxy-5α-cholestan-6-one-disulfate (8). The cytotoxic activity of the steroidal oximes was evaluated against two prostate carcinoma cell lines (PC-3 and LNCaP) and compared with that of five polyhydroxylated sulfated analogs (8-12). Oxime 3 and trisulfated analog 11 were the most active compounds with IC50 values of 10.8µM (PC-3) and 7.9µM (LNCaP), respectively.

Relationship between the 2.4-km run and multistage shuttle run test performance in military personnel.

In the United Kingdom, all branches of the armed forces use 2.4-km run time and/or the 20-m multistage shuttle run test (MSRT) to assess the aerobic fitness of their personnel. This study quantified the relationship between these two tests in 156 army recruits and officer cadets (100 men and 56 women) to ensure equivalence in the required aerobic fitness standards. The 2.4-km run was performed on surfaced roads and tracks around the training establishment and the MSRT in a gymnasium. Ordinary least product regression was used to describe the relationship between average 2.4-km running speed (km · h(-1)) and the total number of shuttles completed on the U.K. version of the MSRT (r = 0.91, p < 0.01), showing MSRT shuttles = (9.708×2.4-km run speed) - 52.56, with a standard error of prediction of approximately 8 shuttles or 0.8 km · h(-1). The British Army 2.4-km run biannual fitness assessment standard for young men of 10:30 min:s equates to a MSRT score of 82 shuttles (level 10 and 1 shuttle) and for young women of 13:00 min:s equates to 56 shuttles (level 7 and 6 shuttles), with a standard error of estimate of approximately 8 shuttles.

Synthesis and acetylcholinesterase inhibitory activity of polyhydroxylated sulfated steroids: structure/activity studies.

Disulfated and trisulfated steroids have been synthesized from cholesterol and their acetylcholinesterase inhibitory activity has been evaluated. In our studies we have found that the activity was not only dependent on the location of the sulfate groups but on their configurations. 2ß,3α,6α-trihydroxy-5α-cholestan-6-one trisulfate (18) was the most active steroid with an IC50 value of 15.48 µM comparable to that of 2ß,3α-dihydroxy-5α-cholestan-6-one disulfate (1). Both compounds were found to be less active than the reference compound eserine. The butyrylcholinesterase activity of 1 and 18 was one magnitude lower than that against acetylcholinesterase revealing a selective inhibitor profile.

Comparison of step-wise and ramp-wise incremental rowing exercise tests and 2000-m rowing ergometer performance.

PURPOSE: This study examined parameters derived from both an incremental step-wise and a ramp-wise graded rowing exercise test in relation to rowing performance. METHODS: Discontinuous step-wise incremental rowing to exhaustion established lactate threshold (LT), maximum oxygen consumption (VO(2maxSTEP)), and power associated with VO(2max) (W VO(2max)). A further continuous ramp-wise test was undertaken to derive ventilatory threshold (VT), maximum oxygen consumption (VO(2maxRAMP)), and maximum minute power (MMW). Results were compared with maximal 2000-m ergometer time-trial power. RESULTS: The strongest correlation with 2000-m power was observed for MMW (r = .98, P < .001), followed by W VO(2max) (r = .96; P < .001). The difference between MMW and W VO(2max) compared with the mean of MMW/W VO(2max) showed a widening bias with a greater difference coincident with greater power. However, this bias was reduced when expressed as a ratio term and when a baseline VO2 was accounted for. There were no differences (P = .85) between measures of VO(2maxSTEP) and VO(2maxRAMP); rather, the measures showed strong association (r = .97, P < .001, limits of agreement = -0.43 to 0.33 L/min). The power at LT and VT did not differ (P = .6), and a significant association was observed (r = .73, P = .001, limits of agreement = -54.3 to 20.2 W, SEE = 26.1). CONCLUSIONS: This study indicates that MMW demonstrates a strong association with ergometer rowing performance and thus may have potential as an influential monitoring tool for rowing athletes.

Development of an accelerometer-based multivariate model to predict free-living energy expenditure in a large military cohort.

This study developed a multivariate model to predict free-living energy expenditure (EE) in independent military cohorts. Two hundred and eighty-eight individuals (20.6 ± 3.9 years, 67.9 ± 12.0 kg, 1.71 ± 0.10 m) from 10 cohorts wore accelerometers during observation periods of 7 or 10 days. Accelerometer counts (PAC) were recorded at 1-minute epochs. Total energy expenditure (TEE) and physical activity energy expenditure (PAEE) were derived using the doubly labelled water technique. Data were reduced to n = 155 based on wear-time. Associations between PAC and EE were assessed using allometric modelling. Models were derived using multiple log-linear regression analysis and gender differences assessed using analysis of covariance. In all models PAC, height and body mass were related to TEE (P < 0.01). For models predicting TEE (r (2) = 0.65, SE = 462 kcal · d(-1) (13.0%)), PAC explained 4% of the variance. For models predicting PAEE (r (2) = 0.41, SE = 490 kcal · d(-1) (32.0%)), PAC accounted for 6% of the variance. Accelerometry increases the accuracy of EE estimation in military populations. However, the unique nature of military life means accurate prediction of individual free-living EE is highly dependent on anthropometric measurements.

Physiological responses of Police Officers during job simulations wearing chemical, biological, radiological and nuclear personal protective equipment.

The aim of this study was to quantify the physiological responses of Police Officers wearing chemical, biological, radiological and nuclear personal protective equipment (CBRN PPE) during firearms house entry (FE) unarmed house entry (UE) and crowd control (CC) simulations. Participants volunteered from the UK Police Force [FE (n = 6, age 33 ± 4 years, body mass 85.3 ± 7.9 kg, (·)VO2max 53 ± 5 ml · kg⁻¹ · min⁻¹), UE and CC (n = 11, age 34 ± 5 years, body mass 88.5 ± 13.8 kg, (·)VO2max 51 ± 5 ml · kg⁻¹ · min⁻¹)]. Heart rate reserve (HRR) during FE was greater than UE (74 ± 7 vs. 62 ± 6%HRR, p = 0.01) but lower in CC (39 ± 7%HRR, p < 0.01). Peak core body temperature was greater during FE (39.2 ± 0.3°C) than UE (38.9 ± 0.4°C, p < 0.01) and CC (37.5 ± 0.3°C, p < 0.01), with similar trends in skin temperature. There were no differences in the volume of water consumed (1.13 ± 0.44 l, p = 0.51) or change in body mass (-1.68 ± 0.65 kg, p = 0.74) between simulations. The increase in body temperature was a primary physiological limitation to performance. Cooling strategies and revised operating procedures may improve Police Officers' physical performance while wearing CBRN PPE. PRACTITIONER SUMMARY: In recent years, the likelihood of Police Officers having to respond to a chemical, biological, nuclear or radiological (CBRN) incident wearing personal protective equipment (PPE) has increased. Such apparel is likely to increase physiological strain and impair job performance; understanding these limitations may help improve Officer safety and operational effectiveness.