{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T19:12:37Z","timestamp":1788462757775,"version":"build-2803163510"},"reference-count":73,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2024,12,9]],"date-time":"2024-12-09T00:00:00Z","timestamp":1733702400000},"content-version":"vor","delay-in-days":8,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001343","name":"University of Pretoria","doi-asserted-by":"publisher","award":["608664"],"award-info":[{"award-number":["608664"]}],"id":[{"id":"10.13039\/501100001343","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100016962","name":"Department of Science and Innovation, South Africa","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100016962","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001332","name":"Council for Scientific and Industrial Research, South Africa","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001332","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bpspubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Pharmacology Res &amp; Perspec"],"published-print":{"date-parts":[[2024,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Acetylcholinesterase inhibitors (AChEIs) remain the first\u2010line 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\u00a0vitro AChEI activity, blood\u2013brain 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\u00a0vitro and in silico methods to determine their feasibility as an inhibitor of the AChE enzyme. An in\u2010house 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\u2010permeable pharmacophores and donepezil were examined using a checkerboard AChEI assay. Cytotoxicity of active compounds and their synergistic combinations was assessed in SH\u2010SY5Y neuroblastoma and bEnd.5 cells using the sulforhodamine B assay. Cellular AChEI activity of active in silico BBB\u2010permeable predicted compounds was determined using an SH\u2010SY5Y AChE\u2010based assay. An in\u00a0vitro 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\u2009\u03bcM, with compound\n                    <jats:bold>A51<\/jats:bold>\n                    showing the lowest IC\n                    <jats:sub>50<\/jats:sub>\n                    (0.20\u2009\u03bcM). Five compounds were identified as BBB\u2010permeable, with the donepezil\u2010\n                    <jats:bold>C53<\/jats:bold>\n                    combination at \u00bcIC\n                    <jats:sub>50<\/jats:sub>\n                    exhibiting the strongest synergy (CI\u2009=\u20090.82). Compounds\n                    <jats:bold>A136<\/jats:bold>\n                    and\n                    <jats:bold>C129<\/jats:bold>\n                    , either alone or with donepezil, showed cytotoxicity. Notably, compound\n                    <jats:bold>C53<\/jats:bold>\n                    , both alone and in combination with donepezil, demonstrated high AChEI activity and promising BBB permeability, warranting further investigation.\n                  <\/jats:p>","DOI":"10.1002\/prp2.70043","type":"journal-article","created":{"date-parts":[[2024,12,9]],"date-time":"2024-12-09T06:41:12Z","timestamp":1733726472000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Evaluating Blood\u2013Brain Barrier Permeability, Cytotoxicity, and Activity of Potential Acetylcholinesterase Inhibitors: In\u00a0Vitro and In Silico Study"],"prefix":"10.1002","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4112-5021","authenticated-orcid":false,"given":"L. M.","family":"Maboko","sequence":"first","affiliation":[{"name":"Department of Pharmacology, School of Medicine, Faculty of Health Sciences University of Pretoria  Pretoria South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0497-4473","authenticated-orcid":false,"given":"A.","family":"Theron","sequence":"additional","affiliation":[{"name":"Future Production: Chemicals, Council for Scientific and Industrial Research  Pretoria South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J.\u2010L.","family":"Panayides","sequence":"additional","affiliation":[{"name":"Future Production: Chemicals, Council for Scientific and Industrial Research  Pretoria South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5744-9285","authenticated-orcid":false,"given":"W.","family":"Cordier","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, School of Medicine, Faculty of Health Sciences University of Pretoria  Pretoria South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6070-0234","authenticated-orcid":false,"given":"D.","family":"Fisher","sequence":"additional","affiliation":[{"name":"Department of Medical BioSciences, Faculty of Natural Sciences, Neurobiology Research Group University of Western Cape  Cape Town South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3575-0410","authenticated-orcid":false,"given":"V.","family":"Steenkamp","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, School of Medicine, Faculty of Health Sciences University of Pretoria  Pretoria South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,12,9]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.trci.2018.03.009"},{"key":"e_1_2_11_3_1","volume-title":"World Alzheimer Report 2021: Journey Through the Diagnosis of Dementia","author":"Gauthier S.","year":"2021"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.32481\/djph.2021.09.009"},{"key":"e_1_2_11_5_1","volume-title":"Enzyme Inhibitors Involved in the Treatment of Alzheimer's Disease","author":"Revadigar V.","year":"2014"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.4236\/iim.2015.75022"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pharep.2014.09.004"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1080\/13543776.2021.1874344"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/BPH.14030"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/S41398\u2010020\u20100709\u2010X"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/978\u20103\u2010030\u201041838\u20100_5"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/S12035\u2010020\u201002120\u2010Z"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.4103\/1673\u20105374.259604"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.36838\/v5i5.9"},{"key":"e_1_2_11_15_1","volume-title":"Aducanumab","author":"Padd I. 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