THC shows activity against cultured Plasmodium falciparum.

The FDA approved drug Dronabinol was identified in a previous study applying virtual screening using the haemozoin crystal as a target against malaria parasites. The active ingredient of dronabinol is synthetic tetrahydrocannabinol (THC), which is one of the major cannabinoids from Cannabis sativa. Traditional use of cannabis for malaria fever was reported in the world's oldest pharmacopoeia, dating to around 5000 years ago. In this research we report that THC inhibits ß-haematin (synthetic haemozoin) and malaria parasite growth. Due the psychoactivity of THC, CBD, the other major naturally occurring cannabinoid that lacks the off-target psychoactive effects of THC, was also tested and inhibited ß-haematin but showed only a mild antimalarial activity. To evaluate whether THC inhibit haemozoin formation, we performed a cellular haem fractionation assay that indicated that is not the likely mechanism of action. For the first time, the cannabinoid chemical structure is raised as a new chemical class to be further studied for malaria treatment, aiming to overcome the undesirable psychoactive effects of THC and optimize the antimalarial effects.

Lapatinib, Nilotinib and Lomitapide Inhibit Haemozoin Formation in Malaria Parasites.

With the continued loss of antimalarials to resistance, drug repositioning may have a role in maximising efficiency and accelerating the discovery of new antimalarial drugs. Bayesian statistics was previously used as a tool to virtually screen USFDA approved drugs for predicted ß-haematin (synthetic haemozoin) inhibition and in vitro antimalarial activity. Here, we report the experimental evaluation of nine of the highest ranked drugs, confirming the accuracy of the model by showing an overall 93% hit rate. Lapatinib, nilotinib, and lomitapide showed the best activity for inhibition of ß-haematin formation and parasite growth and were found to inhibit haemozoin formation in the parasite, providing mechanistic insights into their mode of antimalarial action. We then screened the USFDA approved drugs for binding to the ß-haematin crystal, applying a docking method in order to evaluate its performance. The docking method correctly identified imatinib, lapatinib, nilotinib, and lomitapide. Experimental evaluation of 22 of the highest ranked purchasable drugs showed a 24% hit rate. Lapatinib and nilotinib were chosen as templates for shape and electrostatic similarity screening for lead hopping using the in-stock ChemDiv compound catalogue. The actives were novel structures worthy of future investigation. This study presents a comparison of different in silico methods to identify new haemozoin-inhibiting chemotherapeutic alternatives for malaria that proved to be useful in different ways when taking into consideration their strengths and limitations.

EMG-Informed Neuromusculoskeletal Modelling Estimates Muscle Forces and Joint Moments During Electrical Stimulation.

This study implemented an electromyogram (EMG)-informed neuromusculoskeletal (NMS) model evaluating the volitional contributions to muscle forces and joint moments during functional electrical stimulation (FES). The NMS model was calibrated using motion and EMG (biceps brachii and triceps brachii) data recorded from able-bodied participants (n=3) performing weighted elbow flexion and extension cycling movements while equipped with an EMG-controlled closed-loop FES system. Models were executed using three computational approaches (i) EMG-driven, (ii) EMG-hybrid and (iii) EMG-assisted to estimate muscle forces and joint moments. Both EMG-hybrid and EMG-assisted modes were able estimate the elbow moment (root mean squared error and coefficient of determination), but the EMG-hybrid method also enabled quantifying the volitional contributions to muscle forces and elbow moments during FES. The proposed modelling method allows for assessing volitional contributions of patients to muscle force during FES rehabilitation, and could be used as biomarkers of recovery, biofeedback, and for real-time control of combined FES and robotic systems.

Virtual screening as a tool to discover new ß-haematin inhibitors with activity against malaria parasites.

Malaria remains a major public health problem. With the loss of antimalarials to resistance, the malaria burden will likely continue for decades. New antimalarial scaffolds are crucial to avoid cross-resistance. Here, we present the first structure based virtual screening using the ß-haematin crystal as a target for new inhibitor scaffolds by applying a docking method. The ZINC15 database was searched for compounds with high binding affinity with the surface of the ß-haematin crystal using the PyRx Virtual Screening Tool. Top-ranked compounds predicted to interact with ß-haematin were submitted to a second screen applying in silico toxicity and drug-likeness predictions using Osiris DataWarrior. Fifteen compounds were purchased for experimental testing. An NP-40 mediated ß-haematin inhibition assay and parasite growth inhibition activity assay were performed. The benzoxazole moiety was found to be a promising scaffold for further development, showing intraparasitic haemozoin inhibition using a cellular haem fractionation assay causing a decrease in haemozoin in a dose dependent manner with a corresponding increase in exchangeable haem. A ß-haematin inhibition hit rate of 73% was found, a large enrichment over random screening, demonstrating that virtual screening can be a useful and cost-effective approach in the search for new haemozoin inhibiting antimalarials.

Naphthylisoquinoline alkaloids, validated as hit multistage antiplasmodial natural products.

The discovery and development of multistage antimalarial drugs targeting intra-erythrocytic asexual and sexual Plasmodium falciparum parasites is of utmost importance to achieve the ambitious goal of malaria elimination. Here, we report the validation of naphthylisoquinoline (NIQ) alkaloids and their synthetic analogues as multistage active antimalarial drug candidates. A total of 30 compounds were tested, of which 17 exhibited IC50 values <1 µM against drug-sensitive P. falciparum parasites (NF54 strain); 15 of these retained activity against a panel of drug-resistant strains. These compounds showed low in vitro cytotoxicity against HepG2 cells, with selectivity indices of >10. The tested compounds showed activity in vitro against both early- and late-stage P. falciparum gametocytes while blocking male gamete formation (>70% inhibition of exflagellation at 2 µM). Additionally, five selected compounds were found to have good solubility (≥170 µM in PBS at pH 6.5), while metabolic stability towards human, mouse, and rat microsomes ranged from >90% to >7% after 30 min. Dioncophylline C (2a) emerged as a front runner from the study, displaying activity against both asexual parasites and gametocytes, a lack of cross-resistance to chloroquine, good solubility, and microsomal stability. Overall, this is the first report on the multistage activity of NIQs and their synthetic analogues including gametocytocidal and gametocidal effects induced by this class of compounds.

Identification of 2,4-Disubstituted Imidazopyridines as Hemozoin Formation Inhibitors with Fast-Killing Kinetics and In Vivo Efficacy in the Plasmodium falciparum NSG Mouse Model.

A series of 2,4-disubstituted imidazopyridines, originating from a SoftFocus Kinase library, was identified from a high throughput phenotypic screen against the human malaria parasite Plasmodium falciparum. Hit compounds showed moderate asexual blood stage activity. During lead optimization, several issues were flagged such as cross-resistance against the multidrug-resistant K1 strain, in vitro cytotoxicity, and cardiotoxicity and were addressed through structure-activity and structure-property relationship studies. Pharmacokinetic properties were assessed in mice for compounds showing desirable in vitro activity, a selectivity window over cytotoxicity, and microsomal metabolic stability. Frontrunner compound 37 showed good exposure in mice combined with good in vitro activity against the malaria parasite, which translated into in vivo efficacy in the P. falciparum NOD-scid IL-2Rγnull (NSG) mouse model. Preliminary mechanistic studies suggest inhibition of hemozoin formation as a contributing mode of action.

FES-induced co-activation of antagonist muscles for upper limb control and disturbance rejection.

Control systems for human movement based on Functional Electrical Stimulation (FES) have shown to provide excellent performance in different experimental setups. Nevertheless, there is still a limited number of such applications available today on worldwide markets, indicating poor performance in real settings, particularly for upper limb rehabilitation and assistance. Based on these premises, in this paper we explore the use of an alternative control strategy based on co-activation of antagonist muscles using FES. Although co-contraction may accelerate fatigue when compared to single-muscle activation, knowledge from motor control indicate it may be useful for some applications. We have performed a simulation and experimental study designed to evaluate whether controllers that integrate such features can modulate joint impedance and, by doing so, improving performance with respect to disturbance rejection. The simulation results, obtained using a novel model including proprioceptive feedback and anatomical data, indicate that both stiffness and damping components of joint impedance may be modulated by using FES-induced co-activation of antagonist muscles. Preliminary experimental trials were conducted on four healthy subjects using surface electrodes. While the simulation investigation predicted a maximum 494% increase in joint stiffness for wrist flexion/extension, experiments provided an average elbow stiffness increase of 138% using lower stimulation intensity. Closed-loop experiments in which disturbances were applied have demonstrated that improved behavior may be obtained, but increased joint stiffness and other issues related to simultaneous stimulation of antagonist muscles may indeed produce greater errors.