Evaluating Blood–Brain Barrier Permeability, Cytotoxicity, and Activity of Potential Acetylcholinesterase Inhibitors: In Vitro and In Silico Study
ABSTRACT Acetylcholinesterase inhibitors (AChEIs) remain the first‐line treatment for Alzheimer's disease. However, these drugs are largely symptomatic and often associated with adverse effects. This study aimed to evaluate novel pharmacophores for their in vitro AChEI activity, blood–brain bar...
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Wiley
2024-12-01
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| Series: | Pharmacology Research & Perspectives |
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| Online Access: | https://doi.org/10.1002/prp2.70043 |
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| author | L. M. Maboko A. Theron J.‐L. Panayides W. Cordier D. Fisher V. Steenkamp |
| author_facet | L. M. Maboko A. Theron J.‐L. Panayides W. Cordier D. Fisher V. Steenkamp |
| author_sort | L. M. Maboko |
| collection | DOAJ |
| description | ABSTRACT Acetylcholinesterase inhibitors (AChEIs) remain the first‐line treatment for Alzheimer's disease. However, these drugs are largely symptomatic and often associated with adverse effects. This study aimed to evaluate novel pharmacophores for their in vitro AChEI activity, blood–brain barrier (BBB) permeability, and cytotoxic potential, hypothesizing that a combination of AChEIs could enhance symptom management while minimizing toxicity. A library of 1453 synthetic pharmacophores was assessed using in vitro and in silico methods to determine their feasibility as an inhibitor of the AChE enzyme. An in‐house miniaturized Ellman's assay determined acellular AChEI activities, while pharmacokinetic properties were evaluated using the SwissADME web tool. The combinational effects of in silico BBB‐permeable pharmacophores and donepezil were examined using a checkerboard AChEI assay. Cytotoxicity of active compounds and their synergistic combinations was assessed in SH‐SY5Y neuroblastoma and bEnd.5 cells using the sulforhodamine B assay. Cellular AChEI activity of active in silico BBB‐permeable predicted compounds was determined using an SH‐SY5Y AChE‐based assay. An in vitro BBB model was used to assess the effect of compounds on the integrity of the bEnd.5 monolayer. Out of the screened compounds, 12 demonstrated 60% AChEI activity at 5 μM, with compound A51 showing the lowest IC50 (0.20 μM). Five compounds were identified as BBB‐permeable, with the donepezil‐C53 combination at ¼IC50 exhibiting the strongest synergy (CI = 0.82). Compounds A136 and C129, either alone or with donepezil, showed cytotoxicity. Notably, compound C53, both alone and in combination with donepezil, demonstrated high AChEI activity and promising BBB permeability, warranting further investigation. |
| format | Article |
| id | doaj-art-c601a6189c364fd68ffae00f87743cc0 |
| institution | Kabale University |
| issn | 2052-1707 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Pharmacology Research & Perspectives |
| spelling | doaj-art-c601a6189c364fd68ffae00f87743cc02024-12-13T03:28:40ZengWileyPharmacology Research & Perspectives2052-17072024-12-01126n/an/a10.1002/prp2.70043Evaluating Blood–Brain Barrier Permeability, Cytotoxicity, and Activity of Potential Acetylcholinesterase Inhibitors: In Vitro and In Silico StudyL. M. Maboko0A. Theron1J.‐L. Panayides2W. Cordier3D. Fisher4V. Steenkamp5Department of Pharmacology, School of Medicine, Faculty of Health Sciences University of Pretoria Pretoria South AfricaFuture Production: Chemicals, Council for Scientific and Industrial Research Pretoria South AfricaFuture Production: Chemicals, Council for Scientific and Industrial Research Pretoria South AfricaDepartment of Pharmacology, School of Medicine, Faculty of Health Sciences University of Pretoria Pretoria South AfricaDepartment of Medical BioSciences, Faculty of Natural Sciences, Neurobiology Research Group University of Western Cape Cape Town South AfricaDepartment of Pharmacology, School of Medicine, Faculty of Health Sciences University of Pretoria Pretoria South AfricaABSTRACT Acetylcholinesterase inhibitors (AChEIs) remain the first‐line treatment for Alzheimer's disease. However, these drugs are largely symptomatic and often associated with adverse effects. This study aimed to evaluate novel pharmacophores for their in vitro AChEI activity, blood–brain barrier (BBB) permeability, and cytotoxic potential, hypothesizing that a combination of AChEIs could enhance symptom management while minimizing toxicity. A library of 1453 synthetic pharmacophores was assessed using in vitro and in silico methods to determine their feasibility as an inhibitor of the AChE enzyme. An in‐house miniaturized Ellman's assay determined acellular AChEI activities, while pharmacokinetic properties were evaluated using the SwissADME web tool. The combinational effects of in silico BBB‐permeable pharmacophores and donepezil were examined using a checkerboard AChEI assay. Cytotoxicity of active compounds and their synergistic combinations was assessed in SH‐SY5Y neuroblastoma and bEnd.5 cells using the sulforhodamine B assay. Cellular AChEI activity of active in silico BBB‐permeable predicted compounds was determined using an SH‐SY5Y AChE‐based assay. An in vitro BBB model was used to assess the effect of compounds on the integrity of the bEnd.5 monolayer. Out of the screened compounds, 12 demonstrated 60% AChEI activity at 5 μM, with compound A51 showing the lowest IC50 (0.20 μM). Five compounds were identified as BBB‐permeable, with the donepezil‐C53 combination at ¼IC50 exhibiting the strongest synergy (CI = 0.82). Compounds A136 and C129, either alone or with donepezil, showed cytotoxicity. Notably, compound C53, both alone and in combination with donepezil, demonstrated high AChEI activity and promising BBB permeability, warranting further investigation.https://doi.org/10.1002/prp2.70043acetylcholinesterase inhibitionblood–brain barriercytotoxicitysynthetic compounds |
| spellingShingle | L. M. Maboko A. Theron J.‐L. Panayides W. Cordier D. Fisher V. Steenkamp Evaluating Blood–Brain Barrier Permeability, Cytotoxicity, and Activity of Potential Acetylcholinesterase Inhibitors: In Vitro and In Silico Study Pharmacology Research & Perspectives acetylcholinesterase inhibition blood–brain barrier cytotoxicity synthetic compounds |
| title | Evaluating Blood–Brain Barrier Permeability, Cytotoxicity, and Activity of Potential Acetylcholinesterase Inhibitors: In Vitro and In Silico Study |
| title_full | Evaluating Blood–Brain Barrier Permeability, Cytotoxicity, and Activity of Potential Acetylcholinesterase Inhibitors: In Vitro and In Silico Study |
| title_fullStr | Evaluating Blood–Brain Barrier Permeability, Cytotoxicity, and Activity of Potential Acetylcholinesterase Inhibitors: In Vitro and In Silico Study |
| title_full_unstemmed | Evaluating Blood–Brain Barrier Permeability, Cytotoxicity, and Activity of Potential Acetylcholinesterase Inhibitors: In Vitro and In Silico Study |
| title_short | Evaluating Blood–Brain Barrier Permeability, Cytotoxicity, and Activity of Potential Acetylcholinesterase Inhibitors: In Vitro and In Silico Study |
| title_sort | evaluating blood brain barrier permeability cytotoxicity and activity of potential acetylcholinesterase inhibitors in vitro and in silico study |
| topic | acetylcholinesterase inhibition blood–brain barrier cytotoxicity synthetic compounds |
| url | https://doi.org/10.1002/prp2.70043 |
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