{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T18:59:53Z","timestamp":1785437993388,"version":"3.56.0"},"posted":{"date-parts":[[2026,5,5]]},"group-title":"Neuroscience","reference-count":58,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T00:00:00Z","timestamp":1777939200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["R21AG065653"],"award-info":[{"award-number":["R21AG065653"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"award":["R01AG078615"],"award-info":[{"award-number":["R01AG078615"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Florida Department of Health Ed and Ethel Moore Alzheimer\u2019s Disease Research Program","award":["24A07"],"award-info":[{"award-number":["24A07"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,5]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Alzheimer\u2019s disease (AD) is a multifactorial disease with mixed pathologies. Consequentially, drugs targeting multiple pathological processes may offer synergistic benefits. While histone deacetylase (HDAC) inhibitors have demonstrated efficacy in alleviating AD-related pathologies in animal models, the neuroprotective Wnt\/\u03b2-catenin signaling pathway remains compromised in AD brain. CI-994 is a class I HDAC inhibitor containing N-(2-aminophenyl)-benzamide. Our recent studies indicate that CI-994 is also an activator of Wnt\/\u03b2-catenin signaling by stabilizing Wnt co-receptor LRP6. We herein use CI-994 as a scaffold to develop novel potent dual modulators of class I HDACs and Wnt\/\u03b2-catenin signaling for AD therapy. Our lead compound, W2A-28, selectively inhibits class I HDAC1, 2 and 3 with IC\n                  <jats:sub>50<\/jats:sub>\n                  values of 0.51 \u03bcM, 0.68 \u03bcM, and 0.22 \u03bcM, respectively, and shows no inhibitory activities on other HDACs. Furthermore, W2A-28 potently activates Wnt reporter activity with an EC\n                  <jats:sub>50<\/jats:sub>\n                  value of 1.61 \u03bcM in Wnt-3A-expressing HEK293 cells. As expected, activation of Wnt\/\u03b2-catenin signaling by W2A-28 is associated with elevated LRP6 protein level. Importantly, W2A-28 displays excellent microsomal stability in both mouse and human liver microsomal stability assays, alongside high permeability and a lack of active efflux in MDR1-MDCKII models. Critically, W2A-28 treatment significantly enhances histone acetylation, activates Wnt\/\u03b2-catenin signaling, and suppresses tau phosphorylation in AD patient-specific cerebral organoids carrying\n                  <jats:italic>APOE<\/jats:italic>\n                  \u03b54\/\u03b54 or\n                  <jats:italic>APOE<\/jats:italic>\n                  \u03b53\/\u03b54 with\n                  <jats:italic>PSEN1<\/jats:italic>\n                  M146V mutation. Our findings position W2A-28 as a promising multi-target drug candidate for AD therapy.\n                <\/jats:p>","DOI":"10.64898\/2026.04.30.721954","type":"posted-content","created":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T22:35:21Z","timestamp":1778020521000},"source":"Crossref","is-referenced-by-count":0,"title":["Discovery of a CI-994 derivative as a dual modulator of class I HDACs and Wnt\/\u03b2-catenin signaling for Alzheimer\u2019s disease therapy"],"prefix":"10.64898","author":[{"given":"Wenyan","family":"Lu","sequence":"first","affiliation":[{"name":"Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA"},{"name":"Center for Regenerative Biotherapeutics, Mayo Clinic, Jacksonville, FL 32224, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas R.","family":"Caulfield","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eunmi","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suren","family":"Jeevaratnam","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ni","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guojun","family":"Bu","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takahisa","family":"Kanekiyo","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA"},{"name":"Center for Regenerative Biotherapeutics, Mayo Clinic, Jacksonville, FL 32224, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yonghe","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026051022201006000_2026.04.30.721954v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2022.12.032"},{"issue":"1","key":"2026051022201006000_2026.04.30.721954v1.2","first-page":"S107","article-title":"Multifactorial Hypothesis and Multi-Targets for Alzheimer\u2019s Disease","volume":"64","year":"2018","journal-title":"J Alzheimers Dis"},{"key":"2026051022201006000_2026.04.30.721954v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/nrd4360"},{"key":"2026051022201006000_2026.04.30.721954v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.conb.2020.08.011"},{"key":"2026051022201006000_2026.04.30.721954v1.5","doi-asserted-by":"publisher","DOI":"10.1021\/acschemneuro.1c00775"},{"issue":"1","key":"2026051022201006000_2026.04.30.721954v1.6","doi-asserted-by":"crossref","first-page":"158","DOI":"10.2174\/1570159X19666210609160017","article-title":"Understanding the Role of Histone Deacetylase and their Inhibitors in Neurodegenerative Disorders: Current Targets and Future Perspective","volume":"20","year":"2022","journal-title":"Current Neuropharmacology"},{"key":"2026051022201006000_2026.04.30.721954v1.7","first-page":"1025473","article-title":"The role of altered protein acetylation in neurodegenerative disease","volume":"14","year":"2022","journal-title":"Front Aging Neurosci"},{"key":"2026051022201006000_2026.04.30.721954v1.8","doi-asserted-by":"publisher","DOI":"10.1111\/acel.13895"},{"key":"2026051022201006000_2026.04.30.721954v1.9","doi-asserted-by":"publisher","DOI":"10.1038\/npp.2009.197"},{"key":"2026051022201006000_2026.04.30.721954v1.10","doi-asserted-by":"publisher","DOI":"10.1038\/npp.2015.93"},{"key":"2026051022201006000_2026.04.30.721954v1.11","doi-asserted-by":"crossref","unstructured":"Wang C , Shen D , Hu Y , Chen J , Liu J , Huang Y , et al. 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