In the past few decades, owing to healthy habits and general improvements in lifestyle and medication, life expectancy has substantially increased; however, the prominent upward shift in age distribution has increased the prevalence of chronic diseases, including Alzheimer’s disease (AD). AD slowly affects the brain and exhibits clear pathological changes in the hippocampus, the centre of memory and learning (Zhang et al., 2020). In AD, the propensity of neurotoxic proteins to form template or oligomers is higher and accelerates the conversion and aggregation of endogenous proteins, which eventually convert into fibrils (Schaffert and Carter, 2020). It can be sporadic or familial and AD cases are sporadic in most instances (Dorszewska et al., 2016). Disease modifying treatments primarily focused on reducing amyloid beta (senile plaques, Aβ) and tau (neurofibrillary tangles) load in the brain (Cammisuli et al., 2022). Despite many costly clinical trials ranging from pharmacological to hormonal treatments and immunotherapy, not even a single drug produced clinically significant results due to suboptimal dosing of drugs, unavailability of reliable biomarkers for early diagnosis and more specifically lack of detailed mechanistic approaches (Lashley et al., 2018; Loewenstein, 2022). The existing medication exert only moderate reduction of symptoms; therefore, AD remains symptomatic and can be controlled and prevented but uncured (Fernández and Ribeiro, 2018).

According to the World Alzheimer Report (2018), there are 50 million people living with dementia worldwide, of which 70–80 percent are AD patients, and by 2050 these numbers will be more than triple to 152 million (Patterson, 2018). From the data provided by the World Health Organization (WHO), it is an epidemic worldwide and has become a global burden (Cao et al., 2020). Death from AD has increased 123 percent between 2000–2005 and more than 60 percent cases are from low to middle income countries (Patterson, 2018). At the beginning of 21st century, AD remains a major biomedical challenge. Pharmaceutical companies and neurobiologists around the world are doing their efforts to develop novel FDA approved drugs such as acetyl cholinesterase (AChE) inhibitors (Donepezil, Rivastigmine and Galantamine) and NMDA (n-methyl D aspartate) receptor antagonist (Memantine) but they showed several side effects in phase II and III clinical trials. Common adverse effects of AChE inhibitors are diarrhea, nausea, vomiting, bradycardia, muscle twitching nightmares, etc., and memantine includes dizziness, headache, and lethargy (Ettcheto et al., 2018; Schneider, 2022).

The discovery of new natural pharmacologically active compounds is a widely growing field, as the synthesis of most the biomolecules is tough task (Ramawat and Arora, 2021). Consumption of antioxidant rich food and vegetables might improve brain function, minimize the possibilities of cognitive impairment, retard the process of aging, subsequent oxidation, and disease progression (Andrade et al., 2019). It is clinically proven that they enhance cellular metabolism and nourish brain cells; this safeguarding impact is more potent when isothiocyanates (ITCs) rich fruits and vegetables are specifically consumed (Esteve, 2020; Kamal et al., 2022). The propitious attributes of fruits and vegetables are related to their nutritional and functional components like minerals, vitamins, antioxidants and polyphenols. All of these molecules are found in cruciferous vegetables, however, the sulfurous compound GLCs that give them their distinctive pungent aroma and flavour set them apart. GLCs are stable chemically but biologically inactive and remain sequestered within plant compartment (Verkerk et al., 2009; Alexandre et al., 2020). Tissue damage and chewing are the main causes that lead to the formation of biologically active derivatives of GLCs such as ITCs by enzyme hydrolysis, which directly and indirectly regulate their activity and have been demonstrated to exert neuroprotective properties through multiple mechanisms (Tian et al., 2018).

Generally, there are three major hypothesis, i.e., AChE, amyloid, and tau, which are primarily implicated in Alzheimer’s disease management and prevention. Beside them, neuroinflammation is another important response target involving biochemical events activating resident cells of the central nervous system (CNS), which may induce the entire process of AD. It is initiated by aberrant astrocytes and microglial activation, which leads to the release of different inflammatory mediators such as nitric oxide (NO), prostaglandin E2 (PGE-2), reactive oxygen species (ROS), cytokines and chemokines (Kraft and Harry, 2011). Furthermore, it elevates the level of proinflammatory cytokines such as tumor necrosis factor (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6), which are responsible for neuronal death (Xia et al., 2015). Controlling microglia and astrocytes activation can therefore be a therapeutic approach in the prevention and management of AD. Recently, it has been shown that ITCs possess neuroprotective effects through the modulation of different signalling pathways (Latronico et al., 2021). In oxidative stress and inflammation control, nuclear factor-kβ (NF-kβ) and nuclear erythroid related factor 2 (Nrf2) are two main regulators (Fão et al., 2019). They may primarily be attributed to their peculiar ability to activate the Nrf2/ARE pathway (Giacoppo et al., 2015). ITCs significantly decrease NF-kβ translocation with the inhibition of proinflammatory cytokines (Latronico et al., 2021). Hydrogen sulphide (H₂S) is another important signal molecule in CNS; it could represent an intriguing strategy for the treatment of neurodegenerative diseases (Tabassum and Jeong, 2019; Sharif et al., 2023). Beside this, it also play a key role in many aspects of human health like in antiproliferation, cardioprotection, chemoprevention, etc. (Martelli et al., 2020). It also interacts with redox system regulating cellular oxidative stress and ROS (Kabil and Banerjee, 2010). There is a strong relationship between H₂S and aging, as consistent significant decline of H₂S levels has been observed in AD patients (Disbrow et al., 2021). H₂S is a relevant player accounting for different biophysiological effects of Brassicaceae plants, for example, Allyl isothiocyanate (AITC) from black mustard (B. nigra), benzyl-ITC from garden cress (Lepidium sativum), erucin form Eruca sp., B. oleirecia, etc. and 4-hydorxybenzyl-ITC from white mustard (B. alba) are some important naturally occurring ITCs. Among these selected ITCs, benzyl ITC is the most effective H₂S donor, exhibiting remarkable H₂S release followed by AITC (Citi et al., 2014). Recently, available literature clearly demonstrated that the role of natural ITCs as H₂S donor (Martelli et al., 2020). It is a pleiotropic mediator that affects different element in inflammatory cascade specially NF-kβ and Nrf2 signalling (Zhao et al., 2023).

Another important effect of ITCs is apoptotic suppression as they can intervene and arrest the mitochondrial apoptotic pathway (Dinkova-Kostova and Kostov, 2012). Deposition of Aβ and hyperphosphorylated tau proteins is a crucial event in AD as pathology several studies demonstrated the pharmacological potencies of ITCs against these two hallmarks and their toxicity by intervene in its cascade such as APP cleavage, BACE1 expression, oligomerization of seeded proteins, phosphorylation and dephosphorylation assembly, etc. (Morroni et al., 2018; Asif et al., 2022). ITCs could therefore be considered as a promising source of medicine and for the treatment and management of AD. This review focuses on the knowledge regarding the direct and indirect mechanisms by which GLCs-derived ITCs intervene in inhibition of AChE, neurotoxic proteins (Aβ and tau) and neuroinflammation cascade.

Neurons are the building blocks of the CNS, incapable of reproducing and replacing themselves. Several pathological disorders including AD are caused by the accumulation of reactive oxygen species (ROS) in cells (Deshmukh et al., 2017). The ability of a compound to possess anti-inflammatory, antioxidative, and/or antiapoptotic properties is currently used to establish neuroprotective and neuroinflammatory functions (Dinkova-Kostova and Kostov, 2012). ITCs were reported to play a protective effect in acute and chronic AD (Kamal et al., 2022). A variety of ITCs have been experimentally proven (Table 2) to reduce oxidative stress, inflammation, excitotoxicity, misfolded proteins, and mitochondrial dysfunction, and prevent programmed cell death (Connolly et al., 2021). Through the activation of ARE (antioxidant response element) driven genes, ITCs are strong Nrf-2 (nuclear factor erythroid factor 2) activators. They strongly suppress inflammation via NF-kβ (nuclear factor kappa light chain enhancer of activated β cells) pathway (Sita et al., 2016).

A deficient and non-equilibrium cholinergic neurotransmission is responsible for the pathophysiology of learning and memory resulting behavioral disturbance, progressive loss of cognition and daily routine function (Hoyer, 2004; Craig et al., 2011). In context with the cholinergic hypothesis, decreasing the amount of acetylcholine in the hippocampus and cerebral cortex leads to the dysregulation of ChAT and premature loss of basal forbidden cholinergic neurons (Burčul et al., 2018; Hampel et al., 2019). One of the most significant properties of ITCs is AChE inhibition implicated in acetylcholine neurotransmission (Figure 3). In one study, 11 different ITCs were evaluated for their AChE inhibitory and anti - inflammation properties; the most promising inhibitory activity among 11 ITCs was demonstrated by phenyl isothiocyanate and its derivatives. The most potent AChE inhibitory activity was shown by 2-methoxyphenyl ITC with IC₅₀ value of 0.57 mM. Human COX-2 enzyme was also used to evaluate the anti-inflammatory activity, ranking phenyl ITC and 2-methoxy phenylITC as the most potent with 99% inhibition at 50 μM (Burčul et al., 2018). Moringine-specific benzyl ITC from Moringa Oleifera modulated the Nrf2/AER pathway, proinflammatory biomarkers, and apoptotic pathway in different mouse and rat models (Galuppo et al., 2014, (Galuppo et al., 2015). In another mouse model (LPS induced), it was found that ITCs effectively decreased TNF-α, IL-1β, IL-6 and inhibited NF-kβ (Sailaja et al., 2022). It also downregulated senescence as it promoted neuronal repair in in vitro Aβ induce SH5Y5Y cells (Silvestro et al., 2021).

Through different mechanisms (explained in Table 2), SFN prevented cognitive impairment, reduced the Aβ and tau biomarkers, oxidative stress, inflammation and neurodegeneration in experimental models (Kim, 2021). SFN was able to improve spatial and contextual memory through the Y-maze test and counteract the Aβ aggregate induced memory deficits induced by intracerebroventricular (ICV) injection in a mouse model (Kim, 2021). In the hippocampus and frontal cortex, SFN increased cholinacetyltransferase (ChAT) expression, decreased acetylcholine esterase (AChE) activity, and raised the level of acetylcholine (AChE) (Lee et al., 2014). In another study on a transgenic AD mouse model, it was observed that SFN not only reduced the production and deposition of Aβ plaques in the hippocampus and cerebral cortex but also it is associated with neurobehavioral deficit (Zhang et al., 2015; 2017). The neuroinflammatory inhibition is through the activation of Nrf2/HO-1 pathway and inhibition of JNK/AP-1/NF-Kβ by SFN. SFN significantly increased proteasome activity and enhance Nrf-2 pathway in murine neuroblastoma cell lines (Park et al., 2009). It also modulated the Nrf2/ARE pathway in an AlCl₃-and D-galactose induced mice (Zhang et al., 2014).

Neurogenesis has been shown to be enhanced by AITC and PEITC. AChE inhibitory activity in AD revealed that PEITC inhibited the enzyme more effectively than benzyl ITC and AITC (Burčul et al., 2018). In another study, PEITC inhibited Akt activation, suppressed NO production through INF induction, and had an anti-inflammatory effect (Okubo et al., 2010). PEITC showed a protective effect by modulating the MAPK pathway (Ma et al., 2017). Experimental findings revealed that in LPS-induced inflammation model, AITC showed a neuroprotective effect mediated through downregulation of JNK/NF-kβ/TNF-α signaling (Subedi et al., 2017). It also activated ROS-dependent TRPA1 signaling, resulting in neurogenic inflammation reduction in cultured Schwann cells in vitro (De Logu et al., 2022a; De Logu et al., 2022b). PEITC decreased inflammation and activated the cytoprotective pathway in transgenic mice model by modulating Nrf2/AER pathway and restoring Nrf-2 expression (Boyanapalli et al., 2014; Dayalan Naidu et al., 2018). In another study using LPS-activated rat astrocyte culture, PEITC significantly downregulated MAPK/ERK signaling and influenced the inflammatory pathway (Latronico et al., 2021). Increasing evidences suggests that cytochrome p450 is fundamental for brain homeostasis and function while phase II enzyme such as glutathione S-transferase play a key role in redox homeostasis. Modulation of these enzymes can be achieved by ITCs, in the recent studies glucuronosyltranseferase expression increase by sulforaphane in HepG2 cells, in another study erucin and phenethyl ITC elevated glucuronosyltranseferase activity in rat liver slices (Abdull Razis and Mohd Noor, 2013).

Moringa oleifera extract (MOE) decreased the neuritis resulting from naturally occurring cellular injury, with the development of multipolar primary process (Hannan et al., 2014). It also suppressed oxidative stress, MDA, nitrite and TNF-α, increased SOD and inflammation and improved spatial memory and cholinergic neurotransmission by reducing AChE activity and loss of cortico-hippocampus neurons in rat model fed with M. oleifera seeds in dose dependent manner (Onasanwo et al., 2021). Moringa oleifera extract also scavenged free radicals produced by NO, iNOS and nitrotyrosine increase Nrf2 in LPS-activated macrophages and downregulated antioxidative genes; HO-1, GST-P1 and NQO- (Jaja-Chimedza et al., 2017). In another study, it significantly inhibited AChE and reduced neurodegeneration in an NDD/Al - induced temporocortical degenerated mice model (Ekong et al., 2017). Moringa oleifera - supplemented male Wistar rats showed improved memory when evaluated by the Morris water Maze test and significantly reduced AChE levels in brain tissues in a dose-dependent manner (Adebayo et al., 2021). In another observation, GMC-ITC treated neuronal cells (SH-SY5Y) significantly alleviate oxidative stress condition by reducing ROS level ((Jaafaru et al., 2019a). Glucomoringin ITC (GMC-ITC) isolated from M. oleifera seeds abrogated oxidative stress and showed neuroprotective activity against cytotoxic neuroblastoma cells (SH-SY5Y) induced by H₂O₂, gene expression study of detoxifying markers (phase II) by GMC-ITC revealed that all involved genes significantly express themselves. It also decreased the expression of NF-kβ and increased the expression of Ikβ, Nrf2, SOD-1, NQO1 and Nf-kβ respectively (Jaafaru et al., 2019b). Eruca sativa extract (ESE) with a high amount of erucin (ER) prevented cell death and degeneration induced by LPS in NSC-34 motor neurons exposed to LPS-stimulated macrophage cell culture medium by inhibiting FasL (tumor necrosis factor ligand superfamily number 6 expression) and suppressing pro-inflammatory mediators (attenuates TLR4, COX-2 expression of TNF-α level) (Gugliandolo et al., 2018). Erucin decreased inflammation in different cell line models, counteracted proinflammatory marker expression, and balanced Erk1/2, P38, and JNK signaling (Yehuda et al., 2012; Wagner et al., 2015). Indol 3 carbinol (I3C) is another promising candidate found in vegetables; it reduces the free radical production in neuronal cells (Mammana et al., 2019). It also showed the potent radical scavenging activity by chelating already produced free radical species (Giacoppo et al., 2015). In another study, it suppressed the expression of NO, COX-2, and iNOS in the brain, which prevented apoptosis and inflammation by inhibiting NF-kβ and IB phosphorylation (Kim et al., 2014). Furthermore, it decreased BDNF, GHS and increased TNF-α, IL1-β in mice model, it also helped in suppression of neurodegeneration (Huang et al., 2022). In another experiment, researchers explored the antioxidant and anti-inflammatory activity of SFN and ERN as H₂S donor through the combination with rivastigmine in microglia and neuronal cell line (SH-SY5Y). Result revealed that both derivatives show significant antioxidant and anti inflammatory activities in microglial cell line, expression of antioxidant defense protein (GSH) was also induced in neuronal cell line. It significantly decreased the ROS production and NO release in microglial BV-2 cells. Further Erucin exerted a time dependent Nrf2 activation in SH-SY5Y cells (Sestito et al., 2019). When anti-inflammatory effect of erucin was evaluated in LPS-challenged umbilical vein endothelial cells (HUVECs), it significantly prevented the increase of ROS, TNF-α levels and decreased COX-2. It also induced NF-kβ (Ciccone et al., 2022).

The brain of people suffering from Alzheimer’s disease shows remarkable accumulations of two neurotoxic proteins Aβ and tau (Cao et al., 2020). So far, several Alzheimer’s plaque and tau inhibitors from different sources are available they can target different mechanistic steps of fibril formation. One of the inhibitors that are widely used to stop protein aggregation is GLCs derivatives ITCs as they are consumed as a part of our daily diet (Lopez-Rodriguez et al., 2020). In Table 3, we have discussed some of the GLCs derived ITCs, proposed as the potential inhibitor of misfolded Aβ and tau aggregation and their induced toxicity by different mechanisms and modulation of multiple pathways (Figures 4, 5) as described earlier (Grande et al., 2020). Recent investigations suggested that they may directly interact with misfolded proteins during very early stages of the aggregation cascade by binding and stabilizing unfolded proteins and redirecting the aggregation pathways to form amorphous nontoxic fibrils, blocking seeding and further conformational changes that result in neurotoxicity and cell death.

6-(Methylsulfinyl) hexyl isothiocyanate (6-MSITC) from Wasabia japonica was evaluated against amyloidosis in a murine mice model in which 6-MSITC was induced by intra cerebroventricular injection of Aβ_1-42 oligomers. Behavioral analysis revealed that it reduced Aβ1-42 induced memory impairment in hippocampus tissues, increased ROS, and decreased glutathione levels following Aβ_1-42 injection (Morroni et al., 2018). In another study, the authors observed that Aβ_25-35 induced mitochondrial dependent cell death was blocked by SFN through Nrf2-associated manner (Brasil et al., 2023). Clinically, it inhibited Aβ, reduced its burden, and increased the expression of p75NTR in an intransgenic mouse model (Zhang et al., 2015). In another investigation, SFN was found to suppress Aβ deposition, improve cognition, and locomotor function in aluminum and D-galactose-induced mouse model (Zhang et al., 2017). It modulated the Aβ expression related markers followed CDK5 overexpression inhibition in primary neurons, further it reduced Aβ_1-42 induced neurotoxicity and its deposition in TgCRND8-transgenic mice brains. It also suppressed tau phosphorylation at specific sites (Yang et al., 2023). It reduced and altered hyperphosphorylated tau proteins in embryonic hippocampal rat astrocytes under hypoglycaemic condition at Th 181 and Sr 991/202 within astrocytes (Komiskey et al., 2022). It induced NDP52 by Nrf2 and cleared the phosphorylated tauproteins in mice model (Jo et al., 2014).Through high affinity molecular recognition by heteromeric interaction of Aβ plaques, I3C were found to strongly reduce Aβ fibril formation as observed in microscopic examination by TEM analysis (Cohen et al., 2006).

M.oleifera is profoundly used against chronic diseases including AD. Mitochondrial apoptotic genes profile through GMC-ITC pre-treated SH-SY5Y neuronal cells revealed that it protect the cells against oxidative stress via apoptotic pathway, it significantly downregulate the expression of Bax, CASP3, CASP8, CASP9, Apaf-1, cyt-c, p-53 genes and upregulate Bcl2 gene in mitochondrial apoptotic signalling pathway (Jaafaru et al., 2019a). In another study GMC-ITC from the seeds of M. oleifera significantly decreased the expression of BACE1, APP and increased the expression of MAPT tau genes in H₂O₂ induced cytotoxic neuroblastoma cell (SH-SY5Y) (Jaafaru et al., 2019b). It decreases Aβ production and enhance the synaptic proteins in HHcY induced AD model bydown regulating BACE1. It also played crucial role in Ca²⁺ homeostasis, as it deactivated calpain by decreasing intracellular Ca²⁺ resulting cytosolic protease calpain activity reduction in HHcY induced rat model (Mahaman et al., 2018). In another study conducted on MO-ZnONP treated Sprague Dawley rat model it reduced the Aβ accumulation and helped in sustained brain-Zn content (Dahran et al., 2023).

GLCs derived ICTs are important bioactive natural products that are found in many Brassicaceae plants and few plants from other families. In vitro and animal studies have reported their beneficial effects in neuroprotection and they are reported to enhance cellular metabolism, nourish brain cells, and reduce risk factors associated with neurodegeneration. ITCs inhibit inflammatory mediators, oxidative stress, cellular stress signaling, and improve behavioural measures. They also easily cross the blood brain barrier to interact with particular targets implicated in AD pathogenesis. However, there is no sufficient clinical evidence to prove these effects in humans. Future studies should focus to evaluate their pharmacokinetic parameters and effectiveness in humans.