GSK-3β Inhibitory Activity Research Articles

Design, synthesis and biological evaluation of thieno[3,2-c]pyrazol-urea derivatives as potent glycogen synthase kinase 3β inhibitors based on the DFG-out conformation

Glycogen synthase kinase 3β (GSK-3β) is a potential therapeutic target for the treatment of a variety of human diseases. Here, we report the design and synthesis of a series of thieno[3,2-c]pyrazol-urea derivatives and evaluation of their GSK-3β inhibitory activity. Among these analogues, the compound without substitution on terminal phenyl ring (3a) was found to be the most potent GSK-3β inhibitor with an IC50 of 74.4 nM, while substitution on the terminal phenyl (3b–3p) led to decreased potency, independent of the position, size, or electronic properties of the substituents. Kinase selectivity assay revealed that 3a showed good selectivity over a panel of kinases, but was less selective over CDK1, CDK2 and CDK5. Additionally, the pharmacological properties of the synthesized compounds were investigated computationally by the SwissADME and the results showed that most of the compounds have good ADME profiles.

Connecting GSK-3β Inhibitory Activity with IKK-β or ROCK-1 Inhibition to Target Tau Aggregation and Neuroinflammation in Alzheimer's Disease-Discovery, In Vitro and In Cellulo Activity of Thiazole-Based Inhibitors.

GSK-3β, IKK-β, and ROCK-1 kinases are implicated in the pathomechanism of Alzheimer's disease due to their involvement in the misfolding and accumulation of amyloid β (Aβ) and tau proteins, as well as inflammatory processes. Among these kinases, GSK-3β plays the most crucial role. In this study, we present compound 62, a novel, remarkably potent, competitive GSK-3β inhibitor (IC50 = 8 nM, Ki = 2 nM) that also exhibits additional ROCK-1 inhibitory activity (IC50 = 2.3 µM) and demonstrates anti-inflammatory and neuroprotective properties. Compound 62 effectively suppresses the production of nitric oxide (NO) and pro-inflammatory cytokines in the lipopolysaccharide-induced model of inflammation in the microglial BV-2 cell line. Furthermore, it shows neuroprotective effects in an okadaic-acid-induced tau hyperphosphorylation cell model of neurodegeneration. The compound also demonstrates the potential for further development, characterized by its chemical and metabolic stability in mouse microsomes and fair solubility.

Decoding the molecular Symphony: Unravelling neurologically crucial GSK-3 inhibition through 2D QSAR modelling with MLR, PLS, and ANN approaches

This study aimed to decipher the encoded information within the molecular structure of a dataset of GSK-3β inhibitors through a comprehensive quantity structural-activity relationship (QSAR) investigation employing traditional physicochemical descriptors. Various statistical techniques were applied, encompassing linear methods such as Multiple Linear Regression (MLR) and Partial Least Squares (PLS), along with non-linear approaches such as Artificial Neural Networks (ANN). Rigorous validation using diverse statistical tools confirmed the models’ precision and predictability. The model exhibited exceptional statistical relevance, as evidenced by standard parameters: S-value (0.37), F-value (37.17), r (0.93), r2 (0.855), and r2CV (0.78). They assessed predictive power and robustness by involving specific statistical parameters. Key descriptors analyzed, including Verloop L (subs 2), Lipole Z component (whole molecule), and VAMP Dipole Z component, offered crucial insights into their respective contributions. This analysis implies that targeted modifications in the substitution pattern potentially enhance GSK-3β inhibitory activity significantly. The established model clarified how bioactivity depends on structure and offered valuable recommendations for creating new compounds with better inhibitory activity profiles against the GSK-3β enzyme.

Oxazole-4-carboxamide/butylated hydroxytoluene hybrids with GSK-3β inhibitory and neuroprotective activities against Alzheimer's disease

Neuronal cells overexpressing phosphorylated Tau proteins can increase the susceptibility to oxidative stress. Regulation of glycogen synthase-3β (GSK-3β) and reduction of Tau protein hyperphosphorylation, along with alleviation of oxidative stress, may be an effective way to prevent or treat Alzheimer's disease (AD). For this purpose, a series of Oxazole-4-carboxamide/butylated hydroxytoluene hybrids were designed and synthesized to achieve multifunctional effects on AD. The biological evaluation showed that the optimized compound KWLZ-9e displayed potential GSK-3β (IC50 = 0.25 μM) inhibitory activity and neuroprotective capacity. Tau protein inhibition assays showed that KWLZ-9e reduced the expression of GSK-3β and downstream p-Tau in HEK GSK-3β 293T cells. Meanwhile, KWLZ-9e could alleviate H2O2-induced ROS damage, mitochondrial membrane potential imbalance, Ca2+ influx and apoptosis. Mechanistic studies suggest that KWLZ-9e activates the Keap1-Nrf2-ARE signaling pathway and enhances the expression of downstream oxidative stress proteins including TrxR1, HO-1, NQO1, GCLM to exert cytoprotective effects. We also confirmed that KWLZ-9e could ameliorate learning and memory impairments in vivo model of AD. The multifunctional properties of KWLZ-9e suggest that it is a promising lead for the treatment of AD.

Natural Products with BACE1 and GSK3β Inhibitory Activity.

Alzheimer's disease (AD) is a neurodegenerative, progressive, and fatal disorder characterized by marked atrophy of the cerebral cortex and loss of basal forebrain cholinergic neurons. The main pathological features of AD are related to neuronal degeneration and include extracellular deposition of amyloid beta plaques (Aβ plaques), intracellular formation of neurofibrillary tangles (NFTs), and neuroinflammation. So far, drugs used to treat AD have symptomatic and palliative pharmacological effects, disappearing with continued use due to neuron degeneration and death. Therefore, there are still problems with an effective drug for treating AD. Few approaches evaluate the action of natural products other than alkaloids on the molecular targets of β-amyloid protein (Aβ protein) and/or tau protein, which are important targets for developing neuroprotective drugs that will effectively contribute to finding a prophylactic drug for AD. This review gathers and categorizes classes of natural products, excluding alkaloids, which in silico analysis (molecular docking) and in vitro and/or in vivo assays can inhibit the BACE1 and GSK-3β enzymes involved in AD.

Design, synthesis and bioevaluation of 1,2,4-thiadiazolidine-3,5-dione derivatives as potential GSK-3β inhibitors for the treatment of Alzheimer's disease

Tideglusib is a non-competitive GSK-3β inhibitor which contain 1,2,4-thiadiazolidine-3,5-dione moiety, and now mainly used for progressive supranuclear palsy due to the lack of some primary cognitive endpoints and secondary endpoints in a phase IIb trail for Alzheimer's disease. Additionally, insufficient evidence exists to support that there are obvious covalent bonds between Tideglusib and GSK-3β. Targeted covalent inhibition strategy could improve the binding efficiency, selectivity and duration of kinase inhibitors. Based on the above premise, two series of targeted compounds with acryloyl warheads were designed and synthesized. The kinase inhibitory activity of the selected compound 10a with better neuroprotective effect improved 2.7 fold than that of Tideglusib. After the preliminary screening of GSK-3β inhibition and neuroprotective activity, the mechanism action of the selected compound 10a was investigated in vitro and in vivo. The results confirmed that 10a with excellent selectivity among the whole tested kinases could significantly reduce the expressions of APP and p-Tau via increasing the level of p-GSK-3β. The pharmacodynamic assay in vivo showed that 10a could markedly improve the learning and memory functions in AD mice induced by AlCl3 combined with d-galactose. At the same time, the damage of hippocampal neurons in AD mice was obviously reduced. Accordingly, the introduction of acryloyl warheads could increase the GSK-3β inhibitory activity of 1,2,4-thiadiazolidine-3,5-dione derivatives, and the selected compound 10a deserves further research as an effective GSK-3β inhibitor for the potential treatment of AD.

Oxazole-4-Carboxamide/Butylated Hydroxytoluene Hybrids with GSK-3β Inhibitory and Neuroprotective Activities Against Alzheimer's Disease

Oxazole-4-Carboxamide/Butylated Hydroxytoluene Hybrids with GSK-3β Inhibitory and Neuroprotective Activities Against Alzheimer's Disease

Tenuigenin ameliorates cognitive dysfunction in Alzheimer’s disease via hippocampal neurogenesis enhancement

A promising strategy for treating Alzheimer’s disease (AD) is hippocampal neurogenesis enhancement. Tenuigenin (TEN) is a bioactive compound extracted from Polygala tenuifolia that is widely used for treating amnesia in Chinese medicine. However, whether TEN is effective in treating AD through hippocampal neurogenesis is not fully clear. This study aimed to explore the pharmacologic effect and underlying mechanism of TEN on hippocampal neurogenesis and cognitive deficit amelioration in AD. In an in vivo study, TEN administration significantly ameliorated the cognitive decline in APP/PS1 transgenic AD mice via enhancement of hippocampal neurogenesis, which might be attributed to activation of the GSK-3β/β-catenin pathway. Furthermore, an in silico study suggested that TEN might be directly targeted to GSK-3β. Overall, TEN enhanced hippocampal neurogenesis and consequently ameliorated cognitive deficits via GSK-3β/β-catenin pathway activation, indicating that TEN might be a promising novel agent for AD treatment. • Tenuigenin ameliorates cognitive dysfunction in mouse model of Alzheimer’s disease. • Tenuigenin promotes hippocampal neurogenesis. • Tenuigenin shows potential GSK-3β inhibitory activity.

Synthesis, Characterization and in vitro Studies of Some Ethyl 2-Carboxylate-5- monosubstitued 1H-indole Derivatives as Potential GSK-3β Inhibitors

The present work focuses on indole derivatives due to their promising inhibition activity toward GSK-3β. New compounds based on the indole moiety were synthesized via Japp-Klingemann indole synthesis. The structures of the new compounds were elucidated on the basis of their FTIR, 1H NMR, 13C NMR spectral data, GC-HRMS and elemental analysis. The in vitro GSK-3β inhibitory activity of the new compounds was evaluated using a luminance assay technique in terms of IC50. Compound Aii11 showed excellent inhibitory activity. Compounds Aii2, Aii1 and Aii3 presented promising GSK-3β inhibitory activity.

Design, Bio-evaluation and Molecular Dynamics Simulation of Novel GSK-3β Inhibitors.

Glycogen synthase kinase 3 beta (GSK-3β) is considered as a promising drug target for the treatment of Alzheimer's disease (AD). In the present study, two compound libraries were selected for virtual screening based on pharmacophore models of GSK-3β to discover new inhibitors. Nine potential hits were retained for biological investigation and four of these compounds showed GSK-3β inhibitory activity (with the IC50 values in sub-micromolar range on GSK-3β). Compounds 6 and 9 have good safety. They do not have any significant in vitro cytotoxicity against PC12 and SH-SY5Y neuroblastoma cells at concentrations up to 90 μM. Based on the inhibitory activity and druggability properties, compound 8 is the preferred molecule, and it is a promising lead for the development of the GSK-3β inhibitors for reducing the abnormal hyperphosphorylation of tau protein and relieving AD.

Structural-Based Optimizations of the Marine-Originated Meridianin C as Glucose Uptake Agents by Inhibiting GSK-3β.

Glycogen synthase kinase 3β (GSK-3β) is a widely investigated molecular target for numerous diseases, and inhibition of GSK-3β activity has become an attractive approach for the treatment of diabetes. Meridianin C, an indole-based natural product isolated from marine Aplidium meridianum, has been reported as a potent GSK-3β inhibitor. In the present study, applying the structural-based optimization strategy, the pyrimidine group of meridianin C was modified by introducing different substituents based on the 2-aminopyrimidines-substituted pyrazolo pyridazine scaffold. Among them, compounds B29 and B30 showed a much higher glucose uptake than meridianin C (<5%) and the positive compound 4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione (TDZD-8, 16%), with no significant toxicity against HepG2 cells at the same time. Furthermore, they displayed good GSK-3β inhibitory activities (IC50 = 5.85; 24.4 μM). These results suggest that these meridianin C analogues represent novel lead compounds with therapeutic potential for diabetes.

INTRUSION OF GLYCOGEN SYNTHASE KINASE-3Β TO COPE VARIOUS CARDIAC DISORDERS AT MOLECULAR LEVEL

All eukaryotes consist of kinases with a serine/threonine residue called glycogen synthase kinase 3 (GSK-3) which mediates cellular functions by causing phosphorylation of glycogen synthase and regulating glucose metabolism. It establishes disease mechanisms through cell signalling and different transcription factors. Glycogen synthase kinase-3β (GSK-3β) has pharmacological role in cardiac fibrosis, hyperlipidaemia, hyperglycaemia, hyperhomocysteinemia and in case of myocardial reperfusion injury and estrogen deficiency on the heart. The lead compounds were discovered from natural products possessing GSK-3β inhibitory activity. New signalling pathways involving mitochondrion have been investigated for ischemic preconditioning. GSK-3β may bind with mitochondrial protein and mediate mitochondrion function by binding with PI3K-Akt, PGC-1α, HK-II, PKCε subunits of mPTP. The present study explores the structural functionalities of GSK-3β and their contributory role in cardiac disorders and various other diseases. Therefore, GSK-3β is believed to be an imperative target for the discovery and development of newer drugs.

Development of Bisindole-Substituted Aminopyrazoles as Novel GSK-3β Inhibitors with Suppressive Effects against Microglial Inflammation and Oxidative Neurotoxicity.

Development of glycogen synthase kinase-3β (GSK-3β) inactivation-centric agents with polypharmacological profiles is increasingly recognized as a promising therapeutic strategy against the multifactorial etiopathology of Alzheimer's disease (AD). In this respect, a series of disubstituted aminopyrazole derivatives were designed and synthesized as a new class of GSK-3β inhibitors. Most of these derivatives possess GSK-3β inhibitory activities with IC50 values in the micromolar ranges, among which bisindole-substituted aminopyrazole derivative 6h displayed moderate GSK-3β inhibition (IC50 = 1.76 ± 0.19 μM), and alleviative effects against lipopolysaccharide (LPS)-induced glial inflammation in BV-2 cells and glutamate-induced oxidative neurotoxicity in HT-22 cells. Further in vivo studies indicated that compound 6h had potent anti-inflammatory effect, by showing markedly reduced microglial activation and astrocyte proliferation in the brain of LPS-injected mice. Overall, the simultaneous modulation of 6h on multiple dysfunctions of disease network highlights this structural distinctively bisindole-substituted aminopyrazole could be a useful prototype for the discovery of novel therapeutic agents to tackle AD and other GSK-3β associated complex neurological syndromes.

Abstract LB-236: Design, synthesis, and assessment of anticancer properties of pyrazolopyrimidine derivatives as Glycogen Synthase Kinase-3β inhibitors

Abstract Glycogen synthase kinase-3β (GSK-3β) is a serine/threonine kinase with a crucial role in several key cellular functions including tumor cell proliferation and survival. Yet, GSK-3β has emerged as an attractive target for the development of anti-cancer agents. Several small-molecule have been developed as GSK-3β inhibitors. Amongst, several pyrazolopyrimidines based ATP-competitive inhibitors were identified as promising GSK-3β inhibitors. In this work, several pyrazolo[3,4-d]pyrimidine derivatives were designed, synthesized and assessed for their potential anticancer activity. The new compounds are categorized under two templates, Template A and Template B. In these templates two different linkers with varying lengths and electronic properties were used to link additional aromatic ring systems to the core pyrazolopyrimidine scaffold. The aromatic ring systems were designed with different electronic and bonding properties to test their GSK-3β inhibitory activity and consequently their anticancer properties. All compounds under investigation showed GSK-3β inhibitory properties with IC50's ranging from 0.17- 1.01 µM compared to Tideglusib (standard GSK-3β inhibitor) which showed IC50 of 0.22 µM. Furthermore, the compounds were tested for their potential cytotoxicity against two breast cancer cells (MCF-7 and MDA-MB-231) and lung cancer cells (A549). Only one compound from Template B which possesses the longer hydrazide linker and 2,3-dihydroxy phenyl aromatic ring (compound 27), showed considerable cytotoxic effects against MCF-7, MDA-MB-231 and A549 cells (IC50's of 2.4, 2.4 and 2.3 µM, respectively). Compounds 21, 24, 25 and 26 showed weaker cytotoxic effects against cell lines under investigation (IC50's ranged from 29 to 93 µM). DNA content flow-cytometry analysis was undertaken to evaluate the potential effects of compound 27 on the cell cycle distribution of cell lines under investigation. Compound 27 induced significant antiproliferative effect and accumulation of cells in G0/G1-phase after 24 h of exposure. Longer exposure (48 h) to compound 27 resulted in cell cycle arrest in G2/M-phase. Additional assessment for programmed/non-programed cell death using annexin-V/FITC and acridine orange staining coupled with flow-cytometric analysis showed the ability of compound 27 to induce apoptosis as well as autophagy in all cells under investigation. In conclusion, compound 27 represents a promising GSK-3β inhibitor with potential anticancer properties via the induction of cell cycle arrest with subsequent cellular stress and induction of apoptosis and autophagy. Citation Format: Ahmed T. Negmeldin, Mohammed K. AbdElhameid, Dalia Y. Alsaeedy, Fatiha Hammed, Rasha M. Allam, Walid El-Sayed, Ahmed M. Al-Abd. Design, synthesis, and assessment of anticancer properties of pyrazolopyrimidine derivatives as Glycogen Synthase Kinase-3β inhibitors [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr LB-236.

Multifunctional Ligands with Glycogen Synthase Kinase 3 Inhibitory Activity as a New Direction in Drug Research for Alzheimer's Disease.

Alzheimer's disease (AD) belongs to the most common forms of dementia that causes a progressive loss of brain cells and leads to memory impairment and decline of other thinking skills. There is yet no effective treatment for AD; hence, the search for new drugs that could improve memory and other cognitive functions is one of the hot research topics worldwide. Scientific efforts are also directed toward combating behavioral and psychological symptoms of dementia, which are an integral part of the disease. Several studies have indicated that glycogen synthase kinase 3 beta (GSK3β) plays a crucial role in the pathogenesis of AD. Moreover, GSK3β inhibition provided beneficial effects on memory improvement in multiple animal models of AD. The present review aimed to update the most recent reports on the discovery of novel multifunctional ligands with GSK3β inhibitory activity as potential drugs for the symptomatic and disease-modifying therapy of AD. Compounds with GSK3β inhibitory activity seem to be an effective pharmacological approach for treating the causes and symptoms of AD as they reduced neuroinflammation and pathological hallmarks in animal models of AD and provided relief from cognitive and neuropsychiatric symptoms. These compounds have the potential to be used as drugs for the treatment of AD, but their precise pharmacological, pharmacokinetic, toxicological and clinical profiles need to be defined.

Protective Pathways in Ischaemia–Reperfusion: Take the RISK or Lose the Chance

Protective Pathways in Ischaemia–Reperfusion: Take the RISK or Lose the Chance

Potential anti-Alzheimer effects of selected Lamiaceae plants through polypharmacology on glycogen synthase kinase-3β, β-secretase, and casein kinase 1δ

The aerial parts of Lamiaceae plants have been used since ancient times all over the world, especially in Mediterranean cultures, for their medicinal and culinary properties. Although some of them have a historical role in the improvement of cognition and memory, the precise mechanism of these plants remains unclear.This study evaluated the potential anti-Alzheimer effects of some popular cultivated Lamiaceae plants via their inhibitory potential towards glycogen synthase kinase (GSK-3β), β-secretase (BACE-1), and casein kinase 1δ (CK-1δ), enzymes related to AD.Luminometric-based assays were used for the evaluation of GSK-3β and CK-1δ inhibitory activities in which a fluorescence resonance energy transfer (FRET) assay was used for determining BACE-1 inhibitory activity. The major compounds were identified by high pressure liquid chromatography with diode array detection coupled with electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MS/MS).Among the extracts tested, Origanum onites L., O. majorana L., and Mentha piperita L. were the most active ones for GSK-3β inhibitory activity, with IC50 values under 15 μg/mL, while O. majorana and S. officinalis L. were the most active for BACE-1 and O. onites, O. majorana, and Melissa officinalis L. for CK-1δ, with half maximal inhibitory concentration (IC50) values under 250 μg/mL. On the basis of the composition of these active extracts it is apparent that flavonoids, i.e. apigenin, luteolin, and their glycoside derivatives, along with phenolic acids, i.e. rosmarinic acid and salvianolic acids, are the major principles of these active extracts.The outcomes of this study supported the traditional use of Lamiaceae plants for cognitive disorders and highlighted their underlying mechanisms, targeting GSK-3β/BACE-1/CK-1δ together for the first time.

Amaryllidaceae alkaloids from Narcissus pseudonarcissus L. cv. Dutch Master as potential drugs in treatment of Alzheimer's disease

Twenty-one known Amaryllidaceae alkaloids of various structural types and one undescribed alkaloid, named narcimatuline, have been isolated from fresh bulbs of Narcissus pseudonarcissus L. cv. Dutch Master. The chemical structures were elucidated by combination of MS, HRMS, 1D and 2D NMR spectroscopic techniques, and by comparison with literature data. Narcimatuline amalgamates two basic scaffolds of Amaryllidaceae alkaloids in its core, namely galanthamine and galanthindole. All isolated compounds were evaluated for their in vitro acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), prolyl oligopeptidase (POP), and glycogen synthase kinase-3β (GSK-3β) inhibitory activities. The most interesting biological profile was demonstrated by newly isolated alkaloid narcimatuline.

1-Aryl-3-(4-methoxybenzyl)ureas as potentially irreversible glycogen synthase kinase 3 inhibitors: Synthesis and biological evaluation.

Glycogen synthase kinase 3 (GSK-3) has become known for its multifactorial involvement in the pathogenesis of Alzheimer's disease. In this study, a benzothiazole- and benzimidazole set of 1-aryl-3-(4-methoxybenzyl)ureas were synthesised as proposed Cys199-targeted covalent inhibitors of GSK-3β, through the incorporation of an electrophilic warhead onto their ring scaffolds. The nitrile-substituted benzimidazolylurea 2b (IC50 = 0.086 ± 0.023 µM) and halomethylketone-substituted benzimidazolylurea 9b (IC50 = 0.13 ± 0.060 µM) displayed high GSK-3β inhibitory activity, in comparison to reference inhibitor AR-A014418 (1, IC50 = 0.072 ± 0.043) in our assay. The results suggest further investigation of 2b and 9b as potential covalent inhibitors of GSK-3β, since a targeted interaction might provide improved kinase-selectivity.

Polycyclic maleimide-based derivatives as first dual modulators of neuronal calcium channels and GSK-3β for Alzheimer's disease treatment

Current healthcare has significantly increased the average life expectancy, leading to a consequently greater incidence of age-related diseases, such as Alzheimer's disease. Following a multitarget approach, in this paper a series of polycyclic maleimide-based derivatives were designed and synthesized aimed at simultaneously modulate neuronal calcium channels and glycogen synthase kinase 3-beta (GSK-3β), validated targets to combat Alzheimer’ disease. Different structural modifications were performed on the polycyclic scaffold in order to investigate the structure-activity relationships and compound 10 emerged as a promising non-toxic lead compound, endowed with calcium modulating brain-addressed properties and significant GSK-3β inhibitory activity. Moreover, the easily affordable polycyclic core appears as a new appealing privileged structure in medicinal chemistry.