Amilorid'in İlaç Benzerliği, Farmakokinetik Profili, Toksisite Parametreleri ve Anti-Alzheimer Aktivitesinin Belirlenmesi: Ayrıntılı Bir in silico Araştırma

Öz

Amaç: Alzheimer hastalığı (AH), amiloid-β birikimi ve sinaptik disfonksiyon da dahil olmak üzere karmaşık moleküler mekanizmalarla karakterize edilen ilerleyici bir nörodejeneratif bozukluktur. Bu bağlamda, bu çalışma, amilorid ve memantinin ASIC ile etkileşimleri, farmakokinetik ve toksisite ile ilgili özellikleri açısından karşılaştırmalı bir in silico değerlendirmesini yapmayı amaçlamıştır. Materyal ve Metot: Amilorid ve memantinin yapısal ve farmakokinetik özellikleri, moleküler kenetlenme, ADME tahmini ve toksisite profillemesi de dahil olmak üzere in silico yaklaşımlar kullanılarak değerlendirilmiştir. Moleküler kenetlenme analizleri, Molegro Virtual Docker kullanılarak ASIC reseptörüne (PDB ID: 2QTS) karşı gerçekleştirilmiştir. İlaç benzerliği ve farmakokinetik parametreler, yerleşik hesaplama platformlarına dayanarak değerlendirilmiş ve toksisite uç noktaları in silico toksisite modelleri kullanılarak tahmin edilmiştir. Bulgular: Kenetlenme analizi, amiloridin memantine kıyasla ASIC reseptörüne karşı daha yüksek tahmini bağlanma afinitesi sergilediğini göstermiştir. Her iki bileşik de Lipinski'nin beş kuralına uygunluk göstererek kabul edilebilir ilaç benzeri özellikler sergiledi. Bilgisayar ortamında yapılan ADME tahminleri, özellikle kan-beyin bariyeri geçirgenliği açısından farmakokinetik davranışta farklılıklar ortaya koydu; amilorid, memantine kıyasla merkezi sinir sistemine daha az nüfuz etti. Toksisite tahminleri, her iki bileşiğin de farklı toksisite uç noktalarında değişen olasılıklarla karışık güvenlik profillerine sahip olduğunu gösterdi. Sonuç: Mevcut bilgisayar ortamındaki bulgular, amiloridin hesaplamalı koşullar altında memantine kıyasla ASIC reseptörüyle daha olumlu etkileşime girebileceğini düşündürmektedir. Bununla birlikte, kullanılan yöntemlerin tahmine dayalı doğası göz önüne alındığında, bu sonuçlar dikkatli bir şekilde yorumlanmalıdır. Genel olarak, amilorid, Alzheimer hastalığının ASIC ile ilgili mekanizmalarında daha ileri deneysel araştırmalar için potansiyel bir öncü bileşik olabilir.

Anahtar Kelimeler

• Alzheimer hastalığı
• Amilorid
• Asit duyarlı iyon kanalı
• In silico metot

Destekleyen Kurum

Bu çalışma, Tokat Gaziosmanpaşa Üniversitesi Bilimsel Araştırma Koordinasyon Birimi tarafından desteklenmiştir (Hibre No:2025/76)

Etik Beyan

Çalışmamızda in silico yöntemi kullanıldığı için etik komite onayı alınmadı.

Teşekkür

Bu araştırmaya katılan tüm deneycilerin iş birliğini takdir ediyoruz.

The Determination of Druglikeness, Pharmacokinetic Profile, Toxicity Parameters, and Anti-Alzheimer Activity of Amiloride: A Detailed in silico Investigation

Öz

Objective: Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by complex molecular mechanisms, including amyloid-β accumulation and synaptic dysfunction. In this context, the present study aimed to perform a comparative in silico evaluation of amiloride and memantine in terms of their interactions with ASIC, along with their pharmacokinetic and toxicity-related properties. Materials and Methods: The structural and pharmacokinetic properties of amiloride and memantine were assessed using in silico approaches, including molecular docking, ADME prediction, and toxicity profiling. Molecular docking analyses were performed against the ASIC receptor (PDB ID: 2QTS) using Molegro Virtual Docker. Drug-likeness and pharmacokinetic parameters were evaluated based on established computational platforms, and toxicity endpoints were predicted using in silico toxicity models. Results: Docking analysis indicated that amiloride exhibited a higher predicted binding affinity for the ASIC receptor than memantine. Both compounds complied with Lipinski’s rule of five, suggesting acceptable drug-likeness. In silico ADME predictions revealed differences in pharmacokinetic behavior, particularly in blood–brain barrier permeability, with amiloride showing limited central nervous system penetration compared to memantine. Toxicity predictions suggested that both compounds possess mixed safety profiles, with varying probabilities across different toxicity endpoints. Conclusion: The present in silico findings suggest that amiloride may interact more favorably with the ASIC receptor compared to memantine under computational conditions. However, given the predictive nature of the employed methods, these results should be interpreted cautiously. Overall, amiloride may represent a potential lead compound for further experimental investigation in ASIC-related mechanisms of Alzheimer’s disease.

Anahtar Kelimeler

• Acid-sensitive ion channel
• Alzheimer's disease
• Amiloride
• In silico method

Destekleyen Kurum

This study was supported by Tokat Gaziosmanpaşa University Scientific Research Coordination Unit (Grant No:2025/76)

Etik Beyan

Since the in silico method was used in our study, ethics committee approval was not obtained.

Teşekkür

We appreciate the cooperation of all the experimenters who participated in this research.

Kaynakça

1. 1. Forlenza OV, Diniz BS, Gattaz WF. Diagnosis and biomarkers of predementia in Alzheimer's disease. BMC Med. 2010;8:89. doi.org/10.1186/1741-7015-8-89
2. 2. Long JM, Holtzman DM. Alzheimer's Disease: An update on pathobiology and treatment strategies. Cell. 2019;179(2):312-339.
3. 3. Siddappaji KK, Gopal S. Molecular mechanisms in Alzheimer's disease and the impact of physical exercise with advancements in therapeutic approaches. AIMS Neurosci. 2021;8(3):357-389.
4. 4. Mount C, Downton C. Alzheimer's disease: progress or profit? Nat Med. 2006;12(7):780-784. 5. Ke PC, Sani MA, Ding F, et al. Implications of peptide assemblies in amyloid diseases. Chemical Society Reviews. 2017;46(21):6492-6531.
5. 6. Bisaglia M, Venezia V, Piccioli P, et al. Acetaminophen protects hippocampal neurons and PC12 cultures from amyloid β-peptides induced oxidative stress and reduces NF-κB activation. Neurochemistry international. 2002;41(1):43-54.
6. 7. Takeda K, Uda A, Mitsubori M, et al. Mitochondrial ubiquitin ligase alleviates Alzheimer’s disease pathology via blocking the toxic amyloid-β oligomer generation. Communications Biology. 2021;4(1): 192-199.
7. 8. Murakawa-Hirachi T, Mizoguchi Y, Ohgidani M, Haraguchi Y, Monji A. Effect of memantine, an anti-Alzheimer’s drug, on rodent microglial cells in vitro. Scientific Reports. 2021;11(1):6151-6160.
8. 9. Cummings J, Lee G, Nahed P, et al. Alzheimer's disease drug development pipeline. Alzheimer's & Dementia: Translational Research & Clinical Interventions. 2022;8(1):12295. doi.org/10.1002/trc2.12295

9. 10. Robinson DM, Keating GM. Memantine: a review of its use in Alzheimer's disease. Drugs. 2006;66(11):1515-1534.
10. 11. Areosa SA, Sherriff F. Memantine for dementia. Cochrane Database Syst Rev. 2003;(3):3154.
11. 12. Liu, J, Chang L, Song Y, Li H, Wu Y. The role of NMDA receptors in Alzheimer’s disease. Frontiers in neuroscience. 2019;13:43. doi.org/10.3389/fnins.2019.00043
12. 13. Cakir Z, Yildirim C, Buran I, Önalan EE, Bal R. Acid-sensing ion channels (ASICs) influence excitability of stellate neurons in the mouse cochlear nucleus. Journal of Comparative Physiology A. 2019;205(5):769-781.
13. 14. Zha XM. Acid-sensing ion channels: trafficking and synaptic function. Mol Brain. 2013;6:1.
14. 15. Gonzales EB, Sumien N. Acidity and acid-sensing ion channels in the normal and Alzheimer’s disease brain. Journal of Alzheimer's Disease. 2017;57(4):1137-1144.
15. 16. Wal P, Dwivedi J, Wal A, Vig H, Singh Y. Detailed insight into the pathophysiology and the behavioral complications associated with the Parkinson's disease and its medications. Future Journal of Pharmaceutical Sciences. 2022. doi:10.1186/s43094-022-00425-5
16. 17. Dwivedi J, Seemab M, Wal P, et alThe Intricacies of Polypharmacy and Drug Interactions in Schizophrenia Treatment. Curr Drug Metab. 2025;26(9):579-599. doi:10.2174/0113892002382047250930160449
17. 18. Banerjee P, Kemmler E, Dunkel M, Preissner R. ProTox 3.0: a webserver for the prediction of toxicity of chemicals. Nucleic Acids Research. 2024;52(1):513-520.
18. 19. Bitencourt-Ferreira G, Azevedo WF. Molegro virtual docker for docking. Docking screens for drug discovery. 2019;149-167. doi.org/10.1007/978-1-4939-9752-7_10
19. 20. Lipinski CA, Lombardo F, Dominy BW, Feeney PJ. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Advanced drug delivery reviews. 1997;23(1-3):3-25.
20. 21. Zhu Y, Hu X, Wang L, et al. Recent advances in acid-sensitive ion channels in central nervous system diseases. Current Pharmaceutical Design. 2022;28(17):1406-1411.
21. 22. Mango D, Nisticò R. Neurodegenerative disease: what potential therapeutic role of acid-sensing ion channels? Frontiers in Cellular Neuroscience. 2021;15:730641. doi.org/10.3389/fncel.2021.730641
22. 23. Ismael S, Sakata K, McDonald MP, Liao FF, Ishrat T. ER stress associated TXNIP-NLRP3 inflammasome activation in hippocampus of human Alzheimer's disease. Neurochemistry international. 2021;148:105104. doi.org/10.1016/j.neuint.2021.105104
23. 24. Zhang Z, Chen M, Zhan W, et al. Acid-sensing ion channel 1a modulation of apoptosis in acidosis-related diseases: implications for therapeutic intervention. Cell Death Discovery. 2023;9(1):330.
24. 25. Storozhuk M, Cherninskyi A, Maximyuk O, Isaev D, Krishtal O. Acid-sensing ion channels: focus on physiological and some pathological roles in the brain. Current neuropharmacology. 2021;19(9):1570-1589.
25. 26. Zhang Y, Dong D, Zhang J, et al. Pathology and physiology of acid-sensitive ion channels in the bladder. Heliyon. 2024;10(18). doi:10.1016/j.heliyon.2024.e38031
26. 27. Ruan N, Tribble J, Peterson AM, Jiang Q, Wang JQ, Chu XP. Acid-sensing ion channels and mechanosensation. International journal of molecular sciences. 2021;22(9):4810. doi.org/10.3390/ijms22094810
27. 28. Lin YC, Lee CH, Sung JY, Chen CC. Genetic exploration of roles of acid‐sensing ion channel subtypes in neurosensory mechanotransduction including proprioception. Experimental Physiology. 2024;109(1):66-80.
28. 29. Wal P, Dwivedi J, Wal A, Vig H, Singh Y. Detailed insight into the pathophysiology and the behavioral complications associated with the Parkinson’s disease and its medications. 2022;8(33). doi.org/10.1186/s43094-022-00425-5
29. 30. Ahn YH, Tang Y, Illes P. The neuroinflammatory astrocytic P2X7 receptor: Alzheimer’s disease, ischemic brain injury, and epileptic state. Expert Opinion on Therapeutic Targets. 2023;27(9):763-778.
30. 31. Evlanenkov KK, Zhigulin AS, Tikhonov DB. Possible compensatory role of ASICs in glutamatergic synapses. International Journal of Molecular Sciences. 2023;24(16):12974. doi.org/10.3390/ijms241612974
31. 32. Mango D, Nisticò R. Acid-sensing ion channel 1a: a novel target in Alzheimer’s disease? Neural Regeneration Research. 2023;18(2):324. doi:10.4103/1673-5374.346479