Lippia alba (Mill.) N. E. Brown, an aromatic shrub belongs to the Verbenaceae family widely applied in traditional remedies and pharmaceutical applications. The current study aims to analyse the chemical composition of L. alba leaf essential oil (LALEO) using GC-MS and to assess its in vitro biological activities, including anti-oxidant, anti-inflammatory, acetylcholinesterase inhibitory, and tyrosinase inhibitory effects along with in silico molecular docking against Cox 2(Cyclooxygenase-2) and human acetylcholinesterase (AChE). The LALEO was extracted using Clevenger type apparatus. GC-MS analysis identified 26 chemical compounds, with geranial being the most abundant at 26.21%. ABTS (2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) and DPPH (2,2-diphenyl-1-picrylhydrazyl) assays of LALEO demonstrated significant antioxidant potency, with IC50 values of 10.71 μg/mL and 15.67 μg/mL, respectively. The trypsin inhibitory assay revealed strong anti-inflammatory activity IC50 = 16.12 μg/mL, while the acetylcholinesterase (AChE) inhibitory assay demonstrated an IC50 value of 10.07 μg/mL. The anti-tyrosinase activity yielded an IC50 value of 11.22 μg/mL, outperforming the standard kojic acid (12.56 μg/mL). In silico docking revealed that among the tested constituents, only cubebol (-7.8 kcal/mol) exhibited a higher calculated binding affinity for hAChE than the standard galantamine (-7.6 kcal/mol), whereas β-myrcene (-6.7 kcal/mol) showed weaker binding affinity, indicating cubebol's targeted potential against neurodegenerative disorders linked to AChE imbalance. For COX-2, caryophyllene (-6.3 kcal/mol)) and cubebol (-6.2 kcal/mol) demonstrated comparable, though slightly lower, binding affinities relative to the standard sodium diclofenac ((-6.4 kcal/mol), highlighting their potential as preliminary computational lead hits for multi-target virtual screening. The present study suggests that LALEO has potential for further research and the development of natural medicines, after having scientific validation.

Cognitive impairment and anxiety-like behavior associated with cholinergic dysfunction and oxidative stress remain important experimental targets for neuropharmacological screening. Mentha spicata essential oil (MEO, 150 or 300 µL/L) was administered daily by immersion from experimental day 1 through day 22; the first behavioral assessment occurred after 7 days of pre-exposure. Scopolamine hydrobromide trihydrate (SCO, 100 µM) was administered for 30 min before each behavioral assessment and again before euthanasia, whereas galantamine (GAL, 1 mg/L) was administered 30 min before each SCO challenge. Behavior was evaluated using the novel tank diving test (NTT), novel approach test (NAT), Y-maze, and novel object recognition test (NOR). Brain acetylcholinesterase (AChE) activity, antioxidant defenses, and oxidative damage were assessed. GC-MS identified a carvone-rich profile dominated by carvone (67.8%), limonene (10.6%), and 1,8-cineole (4.2%). MEO attenuated several SCO-associated behavioral and biochemical changes. The 150 µL/L concentration showed the most consistent memory-related improvement, whereas 300 µL/L also reduced selected oxidative markers but was associated with reduced exploratory activity in some tasks. Treatment-adjusted behavior-biomarker analyses were considered exploratory; only the inverse association between malondialdehyde (MDA) and the NTT top/bottom ratio remained significant after false-discovery-rate correction. In silico descriptors were treated as hypothesis-generating and do not demonstrate zebrafish brain exposure. These findings support further investigation of carvone-rich MEO within a rescue/protection framework, with confirmation in independently replicated exposure tanks and inclusion of MEO-only controls.

Over the years, our group has used complementary approaches to examine how autonomic-immune crosstalk influences the pathogenesis of systemic lupus erythematosus (SLE). We found that pharmacological enhancement of vagal activity and engagement of the parasympathetically-mediated cholinergic anti-inflammatory pathway via systemic administration of galantamine attenuated pathogenic autoantibodies, blood pressure and inflammatory mediators in murine lupus. With longer administration, galantamine also attenuated splenic B cells and renal fibrosis, and improved survival. Complementary B cell depletion studies in the same mouse model supported an important role for B-cell-dependent autoimmunity in hypertension and kidney injury in lupus, providing mechanistic context for why cholinergic anti-inflammatory pathway-targeted interventions are effective. In contrast, direct localized chemogenetic stimulation of efferent parasympathetic neurons in the dorsal motor nucleus of the vagus did not alter blood pressure or autoantibody levels but improved renal pathology in lupus mice. Similarly, bilateral denervation of renal sympathetic nerves did not affect blood pressure or autoantibody production but reduced albuminuria in lupus mice. Together, these studies demonstrate that direct modulation of autonomic nerve activity can selectively influence kidney outcomes and that a precise balance between parasympathetic and sympathetic signaling is required to maintain immune cell and inflammatory homeostasis. Disruption of this balance may contribute to chronic inflammatory diseases such as SLE and others. This review highlights emerging opportunities for cholinergic anti-inflammatory pathway-based pharmacological and neuromodulatory strategies to mitigate inflammation-driven autoimmune, cardiovascular, and renal disease.

A naive Bayesian classifier (NBC) combined with principal component analysis (PCA) effectively differentiates the dose-dependent effects of certain groups of psychoactive drugs based on their impact on the amplitude-spectral characteristics of electrocorticograms (ECoG) in rats. This approach has been shown to be useful for pharmacological screening of agents with unknown or poorly understood activity. Despite previously obtained optimistic results, classification determination for some drugs was inaccurate, necessitating the search for possible ways to improve the predictive effectiveness of the proposed algorithm. One possible approach would be to use as input quantitative data not only the impact of the psychoactive drugs studied on the amplitude-spectral characteristics of ECoG but also connectivity changes, including the average coherence power of different pairs of leads. The aim of this study was to compare the accuracy of NBC in classifying the pharmacological mechanism of action of agents with well-known mechanisms (test set) using pharmaco-EEG data on changes in the amplitude-spectral characteristics of ECoG, coherence, and the combined use of two data sets. Materials and methods. Experiments were performed on Wistar rats with chronically implanted ECoG electrodes. The training set, relative to which the effects of the pharmacological agents from the test set were classified, were the matrices of effects of 12 pharmacological agents: the NMDA antagonist dizocilpine, the D2/D3 antagonists haloperidol and sulpiride, the M-anticholinergic tropicamide, the H1/5HT2A receptor blocker hydroxyzine, the acetylcholinesterase inhibitor galantamine, the alpha-2 adrenergic agonist dexmedetomidine, the alpha-2 adrenergic antagonist atipamezole, the adenosine receptor blocker caffeine and the GABA-mimetics aminophenylbutyric acid (phenibut), bromdihydrochlorophenylbenzodiazepine (phenazepam) and 5-ethyl-5-phenyl-2,4,6(1H,3H,5H)-pyrimidinetrione. The test set included various drugs with tropism for the targets of the training set drugs: dopamine receptor antagonists chlorpromazine, droperidol, tiapride and raclopride, H1-histamine blockers diphenhydramine and promethazine, 5-HT2-receptor blockers ritanserin and glemenserin, acetylcholinesterase inhibitor ipidacrine, alpha2-adrenergic receptor antagonist yohimbine, alpha2-adrenergic agonists medetomidine and xylazine, GABA-mimetics 5-ethyl-5-(1-methylbutyl)-2,4,6(1H,3H,5H)-pyrimidinetrione and chloral hydrate. The analysis of the ECoG signal included the calculation of 132 amplitude-spectral characteristics and 75 coherence indicators, which, using the PCA, led to new integrative indicators used for further classification of the NBC. Results and discussion. For each drug in the test set, the median similarity probability with a particular group from the training set was calculated, which was used to assess the classification quality. It was found that, when using the amplitude-spectral characteristics of ECoG, the proposed methodological approach allows for the identification of the ECoG effects of several groups of psychoactive drugs, including D2/D3-dopamine, M-cholinergic, H1-histamine, and 5-HT2-serotonin receptor blockers, AChE inhibitors, GABA-mimetics, and alpha-2-adrenergic receptor agonists and antagonists. This approach enabled the correct classification of 18 of 24 groups in the test set. When using changes in coherence indices as the initial data, the classification accuracy also amounted to 18 of 24 groups. When combining the two data sets, the number of correctly identified NBC groups was 20 of 24 groups. When comparing the classification during training (confusion matrix), it was found that coherence data or adding coherence data to the data based on changes in amplitude-spectral characteristics leads to a statistically significant (p < 0.01 in both cases) increase in accuracy. Conclusions. The obtained data demonstrated high accuracy in classifying the pharmacological activity of the test sample drugs using any of the three compared approaches. Despite the lack of statistically significant differences between them, classification based on the combined dataset demonstrated a higher number of "correct" similarities. This allows us to recommend the approach based on combined data of drug effects on amplitude-spectral characteristics and coherence as the most promising for further studies using pharmaco-EEG screening.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline, memory impairment, and neuronal loss, primarily associated with β-amyloid plaque deposition, tau hyperphosphorylation, and cholinergic dysfunction. Since the disruption of cholinergic neurotransmission is a key pathological feature of AD, the cholinesterase (ChE) inhibitors targeting acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) remain the mainstay of symptomatic treatment by enhancing synaptic acetylcholine levels. However, the currently approved small molecules, including tacrine, donepezil, rivastigmine, and galantamine, are limited by their modest efficacy, poor selectivity, and adverse effects, necessitating the development of improved therapeutic agents. Among emerging scaffolds, isatin (1H-indole-2,3-dione) has attracted considerable research attention owing to its structural versatility, synthetic accessibility, and favorable pharmacological properties. Recent studies have reported diverse isatin-based derivatives, including hydrazones, Schiff bases, thiosemicarbazones, spirooxindoles, and molecular hybrids, exhibiting potent AChE and BChE inhibitory activities, often in the submicromolar or nanomolar ranges. Structure-activity relationship studies have revealed the critical influence of electronic effects, linker optimization, N-substitution, and molecular hybridization on inhibitory potency and selectivity. This review summarizes the most promising isatin-based cholinesterase inhibitors reported from 2020 to the present, highlighting their biological activities, structure-activity relationships, and potential for the development of next-generation anti-Alzheimer therapeutics.

Organophosphate pesticides, such as malathion, have been linked to various pathological effects, including neurodegeneration. Acute exposure to malathion, a non-reversible acetylcholinesterase (AChE) inhibitor, causes adverse effects in mammals. However, the impact of subacute, occupational-like repeated exposure on motoneuron (MN) and neuromuscular junction (NMJ) anatomy is less understood. We hypothesized that repeated malathion exposure would produce motor performance deficits accompanied by anatomical alterations in spinal MNs and NMJs consistent with neurodegeneration. We also hypothesized that galantamine, a reversible AChE inhibitor, would provide neuroprotection. Adult male and female Sprague-Dawley rats received subcutaneous saline, malathion (50 mg/kg), galantamine (5.0 mg/kg), or both once daily, five days per week for four weeks. Animals were evaluated immediately after the final exposure and following a four-week recovery period. After exposure, no evidence of spinal astrogliosis and plasma cholinesterase activity was detected. Despite this, cholinergic (C-bouton) synaptic remodeling was observed on lumbar spinal MN somas in females and both sexes showed decreased open-field motor activity. Exposed males performed worse on the accelerod, while exposed females exhibited reduced grip strength. These functional deficits corresponded with a 25% decrease in hindlimb NMJ innervation in both sexes. After recovery, exposed rats showed increased NMJ fragmentation, suggesting remodeling during recovery. Galantamine treatment protected against health effects associated with repeated occupational-like malathion exposure, indicating involvement of cholinergic mechanisms despite minimal changes in plasma cholinesterase activity. Overall, repeated low-concentration malathion exposure is associated with locomotor deficits and peripheral neuropathy.

Several studies have supported Dr Gideon Koren’s 1993 landmark article, “Medications Which Can Kill a Toddler with One Tablet or Teaspoonful.” Although each study has its own merits, most focus solely on pediatric overdoses, and few provide an exhaustive reference list for real-time evaluation of ingestions in children and adults. The following xenobiotics warrant special attention because of their potential to cause serious outcomes, even after exposure to small volumes. This list does not imply that all pediatric exposures to these agents result in fatality, and the absence of a xenobiotic from this list does not indicate that it is necessarily safe. α1-Adrenergic agonists and imidazoline receptor agonists: Clonidine, guanfacine, naphazoline, oxymetazoline, tetrahydrozoline, xylazine (veterinary). Sulfonylureas: Chlorpropamide, glyburide, glipizide, glimepiride. Calcium channel blockers: Verapamil, diltiazem, nifedipine, amlodipine, nicardipine, nimodipine; β-Blockers: Metoprolol, labetalol, propranolol, sotalol. Tricyclic antidepressants: Imipramine, desipramine, amitriptyline, nortriptyline. Opioids: Codeine, hydrocodone, methadone, morphine, heroin. Antidiarrheals: Diphenoxylate plus atropine, loperamide. Salicylates and methyl salicylate: Oil of wintergreen, bismuth subsalicylate, mentholated balms, paramao or lawang root oil . Antipsychotics: Loxapine, thioridazine, chlorpromazine. Antimalarials: Chloroquine, hydroxychloroquine, quinine, primaquine. Antiarrhythmics: Quinidine, disopyramide, procainamide, flecainide, amiodarone. Camphor: A terpenoid used as an analgesic, antipruritic agent, and component of cooling gels, ointments, balms, mothballs, household fragrances, and essential oils. Oral camphor use occurs in some regions for religious ceremonies, traditional remedies, aphrodisiacs, or lactation suppression. Nonalkaloid toxic lignan: Podophyllin, podofilox. Unique plant toxins: Colchicine. Oral acetylcholinesterase inhibitors: Rivastigmine, donepezil, galantamine, carbamate pesticides. Methylxanthines: Theophylline, 1,3-dimethylxanthine, caffeine. Other opioids: Buprenorphine, fentanyl. Toxic alcohols: Methanol, ethylene glycol, diethylene glycol. Caustics and other household products: Acidic or alkaline household products, hydrofluoric acid, selenious acid, acetonitrile, methylene chloride. Accidental overdoses continue to occur regularly despite the implementation of various safety measures, including widespread public education, the use of childproof containers, and easy access to poison control centers. In the US, poison control centers receive more than 2.4 million calls annually, with more than 50% involving children younger than 6 years. Most pediatric accidental ingestions involve household cleaning products and analgesics, followed by cosmetics, foreign objects, and dietary supplements. Adult overdoses may be unintentional but often involve larger total doses or high-risk medications, including opioids and sedative-hypnotics. America's Poison Centers identify the 5 most common substance classes as analgesics, household cleaners, antidepressants, cosmetic or personal care items, and cardiovascular drugs. The number of ingestions involving cleaning products increased sharply during the COVID-19 pandemic but has since declined.

The phytochemical profile and multi-target bioactivity of four Turkish Rosa species, R. canina, R. hemisphaerica, R. horrida, and R. pulverulenta, were evaluated across vegetative organs (twigs and leaves) and three solvent polarities (ethyl acetate (EA), methanol (MeOH), water), in total 24 extracts. Untargeted Ultra-High-Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry (UHPLC-QTOF-MS) metabolomics, along with total phenolic (TPC) and flavonoid (TFC) content, six antioxidant assays and inhibition of seven enzymes were determined. Twig EA extracts displayed unexpectedly high TPC (83-105 mg gallic acid equivalent (GAE) per g) and TFC (35-47 mg rutin equivalent (RE) per g), indicative of selective aglycone flavonoid enrichment. The phenolic profiles identified 337 putative compounds, of which 60.84% belonged to the flavonoid class. The semi-quantitative analysis also indicates that methanol and water extracts had higher quantities of glycosylated flavonoids and phenolic acids, while EA extracts were richer in lignans, especially R. horrida-leaf-EA (1743.25 µg SE per g DW), and polyphenols. In enzyme inhibition assays, EA extracts demonstrated consistent multi-target activity: AChE inhibition (up to 2.49 mg galantamine equivalent (GALAE) per g), BChE inhibition (up to 4.44 mg GALAE per g), tyrosinase inhibition (up to 63.58 mg kojic acid equivalent (KAE) per g), α-amylase inhibition (up to 0.72 mmol acarbose equivalent ACAE per g), and carbonic anhydrase inhibition (CA I IC50 as low as 12.17 µg mL[-1]). The R. hemisphaerica-twig-EA extract showed highly selective cytotoxic activity against A375 cells, with a 50% cytotoxic concentration (CC50) of 3.22 µg mL[-1] and an SI of 73.95, compared to VERO cells. This is the first report of carbonic anhydrase inhibition, BChE-selective activity, cytotoxicity, and comprehensive enzyme-inhibitory data for R. hemisphaerica, R. horrida, and R. pulverulenta. These findings reframe the phytopharmacological potential of Turkish Rosa species beyond their traditionally studied fruit.

The principal constraint on early-stage medicinal chemistry in an academic setting is rarely the supply of chemical ideas, but the cycle time required to convert them into tested compounds. Here we benchmark an integrated platform that couples ensemble virtual screening, automated continuous-flow library synthesis with inline scavenging, and biological profiling. The platform was evaluated deliberately on a target-scaffold combination for which the pharmacology is already established: the N-benzylpiperidine carboxamide class, identified in our earlier virtual screening campaign against acetylcholinesterase (AChE)[1] and structurally anchored to the approved therapeutic donepezil (1). This choice makes platform performance, the measured variable. An 84-member virtual library was designed, triaged by ensemble docking, and synthesized on an automated flow platform; 54 members were isolated in ≥95% purity. Single-point screening at 5 μM identified ten compounds with ≥70% AChE inhibition, and dose-response determination against electric eel AChE (eeAChE) gave three sub-200 nM inhibitors: 69 (91 nM), 8 (93 nM) and 12 (117 nM), each exceeding galantamine (237 nM) and approaching donepezil (1, 42 nM) under identical assay conditions. Cytotoxicity against VERO cells was uniformly low (IC50 > 250 μM), giving selectivity indices above 2000. Molecular dynamics simulations and twelve single-crystal X-ray structures provide a structural basis for the observed structure-activity relationships, identifying a C-Br···O halogen bond to Asp72 as the origin of the ortho-bromo preference, and a binding-mode reversal that accounts for the loss of potency on benzylpiperazine extension. The complete cycle was executed in under three months by a three-person team. We report this as a measured platform capability rather than as accelerated drug discovery allowing the rapid generation of first-round leads.

Alzheimer's disease (AD) is the leading cause of dementia and represents a public health challenge in aging populations. Acetylcholinesterase inhibitors (AChEIs) remain first-line symptomatic therapy for mild to moderate AD, with real-world effectiveness largely influenced by tolerability, adherence, and treatment persistence. Galantamine and its inactive prodrug benzgalantamine are differentiated within the class by a dual mechanism that combines reversible acetylcholinesterase inhibition with positive allosteric modulation of nicotinic acetylcholine receptors. This narrative review synthesizes preclinical and clinical evidence examining galantamine-based therapy, with a focus on aspects related to nicotinic acetylcholine receptors: behavioral symptoms and sleep-related effects. Across randomized, observational, and longitudinal studies, galantamine has been associated with improvement or stabilization in behavioral domains including aberrant motor behavior, agitation/aggression, anxiety, apathy, delusions, disinhibition, and hallucinations. However, like other members of the AChEI class, gastrointestinal tolerability issues may affect treatment persistence. Benzgalantamine was developed to address gastrointestinal tolerability, with the aim of improving adherence and long-term treatment continuity.

In this study, bis-Schiff base derivatives bearing 4-hydroxyacetophenone were synthesized and assessed for their in vitro acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitory activities. In the series, compound (2i) was identified as the most promising inhibitor of AChE (IC50 = 14.82 ± 0.09 µM) and BuChE (IC50 = 27.91 ± 0.16 µM) enzymes surpassing the standard drug galantamine, while the remaining derivatives showed good to moderate inhibition. Structure-activity relationship (SAR) study exposed those methoxy groups attached to the benzene ring significantly improved enzyme inhibition, while electron-withdrawing groups led to condensed the activities. Molecular docking simulations against AChE and BuChE revealed distinct binding modes, with 2i forming critical bonds with catalytic residues. Quantum chemical analysis established strong correlations between electrophilicity index (ω = 4.712 eV for 2i), HOMO-LUMO gap (4.287 eV), and experimental inhibition. Quantum Theory of Atoms in Molecules (QTAIM) analysis supporting optimal electron density distribution and favorable non-covalent interaction patterns. Comprehensive in silico ADMET profiling predicted excellent pharmacokinetic properties for all derivatives, with 96%-100% human oral absorption, moderate blood-brain barrier (BBB) penetration for 2i, and low cardiac toxicity risk. The integrated computational-experimental approach provides supportive evidence for rational design of next-generation cholinesterase inhibitors with optimized efficacy and safety profiles.