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Multi-Target Neuroprotective Compound Exhibits EAAT2-Modulating and Alzheimer's Pathology-Attenuating Effects in In Vitro and In Vivo Models.

Overview

Authors: Ahmet Hacımüftüoğlu1, Nurullah Saraçoğlu2,3, Sana Saffour4, Nadeem Abad4, Yunus Kesgun4, Nadjiba Zegheb4, Ersin Gundeger5, Fatma Yeşilyurt1, Merve Nur Ataş4, Gizem Bati-Ayaz4, Öznur Altunlu1, Burak Çınar1, Mehmet Ali Yörük1, Ufuk Okkay1, Mustafa Özkaraca6, Orhan Ateş7, Ali Taghizadehghalehjoughi8, Ferruh Lafzi2, Hasan Türkez9
  1. Department of Medical Pharmacology, Faculty of Medicine, Atatürk University, 25240 Erzurum, Turkey
  2. Department of Chemistry, Faculty of Sciences, Atatürk University, 25240 Erzurum, Turkey
  3. Biotechnology Institute, Ankara University, 06135 Ankara, Turkey
  4. Trustlife Laboratories Drug Research & Development Center, 34774 İstanbul, Turkey
  5. Biochemistry Department, Faculty of Pharmacy, Istanbul Health and Technology University, Sütlüce, Beyoğlu, 34275 İstanbul, Turkey
  6. Faculty of Veterinary Medicine, Department of Pathology, Cumhuriyet University, 58140 Sivas, Turkey
  7. Department of Eye Diseases, Faculty of Medicine, Atatürk University, 25240 Erzurum, Turkey
  8. Department of Medical Pharmacology, Faculty of Medicine, Bilecik Seyh Edebali University, 11230 Bilecik, Turkey
  9. Department of Medical Biology, Faculty of Medicine, Atatürk University, 25240 Erzurum, Turkey
Journal: ACS chemical neuroscience, volume 17, issue 10, pages 1919-1944
Dates: received 6 November 2025; accepted 9 April 2026; published online 29 April 2026; in print May 2026
Type: Research article · Language: English
License: CC BY
Identifiers: DOI 10.1021/acschemneuro.5c00873 · PMID 42051019 · PMCID PMC13195668 · OpenAlex W7158231128
Open access: hybrid, a free copy (OpenAlex)
Status: data only
Categories: rat (organism), Alzheimer's / dementia (population), cellular / molecular (subfield)
Methods: Statistics, Connectivity
Keywords: neurodegeneration, glutamate excitotoxicity, Tau pathology, β-Amyloid pathology, oxidative stress
MeSH: Alzheimer Disease*, Excitatory Amino Acid Transporter 2*, Neuroprotective Agents*, Animals, Astrocytes, Cell Survival, Cells, Cultured, Disease Models, Animal, Donepezil, Female, Glutamic Acid, Hippocampus, Neurons, Rats, Rats, Sprague-Dawley (* major topic)
Topic: Alzheimer's disease research and treatments (Physiology, Medicine), according to OpenAlex
Citations: not cited yet (Europe PMC); 103 references in the paper

Abstract

Alzheimer’s disease (AD) is a debilitating neurodegenerative disorder characterized by cognitive decline and memory loss. Current treatments offer limited efficacy, necessitating the development of innovative multitarget therapeutic strategies. Here, we present N 3,N 5-bis­(2-(5-methoxy-1H-indol-3-yl)­ethyl)-2,6-dimethyl-4-(2-nitrophenyl)­pyridine-3,5-dicarboxamide (HCM-01), a novel compound developed to target multiple neurodegenerative pathways implicated in AD. In vitro assays included MTT-based cell viability analyses performed in two complementary experimental settings: primary neuronal cultures and astrocyte-based in vitro cell culture models exposed to glutamate. In primary hippocampal neuronal cultures, glutamate exposure induced a statistically significant reduction in cell viability compared with vehicle-treated controls, consistent with glutamate-induced excitotoxicity. Under these conditions, HCM-01 treatment resulted in a statistically significant improvement in neuronal viability, showing a greater protective effect compared with donepezil and memantine. In contrast, in astrocyte-based in vitro cultures, the applied glutamate concentration did not induce overt cytotoxicity, in line with the intrinsic neuroprotective and glutamate-buffering role of astrocytes. Accordingly, astrocytic experiments were designed to assess functional modulation of glutamate-handling mechanisms rather than cell survival. Western blot analysis in C8-D1A astrocytic cells demonstrated increased expression of excitatory amino acid transporter 2 (EAAT2) following HCM-01 treatment compared with control and reference drug-treated groups, suggesting modulation of astrocyte-mediated glutamate homeostasis. In parallel, redox analyses revealed that HCM-01 improved oxidative/antioxidative balance, as evidenced by increased total antioxidant capacity (TAC) and reduced total oxidant status (TOS), supporting an indirect antioxidant contribution to its functional effects. In vivo behavioral assessment of HCM-01 in a streptozotocin (STZ)-induced Alzheimer’s model in female Sprague–Dawley rats demonstrated that administration of HCM-01 at doses of 50 mg/kg orally (oral, P.O. and intraperitoneal, I.P.) and 100 mg/kg (P.O.), significantly improved cognitive and memory functions in the passive avoidance (PA), Morris water maze (MWM), and locomotor activity tests. Moreover, histopathological and immunohistochemical analyses of different hippocampal regions revealed reduced neuronal damage, attenuation of tau pathology, antiamyloidogenic effect, and restoration of cholinergic function. Complementary in silico studies, including molecular docking, molecular dynamics simulations (MDS), and free energy calculations, suggested potential interactions of HCM-01 with the allosteric site of EAAT2. Taken together, these findings suggest that HCM-01 exerts neuroprotective effects against glutamate-induced excitotoxicity in primary hippocampal neurons while additionally modulating glutamatergic homeostasis and redox balance through functional mechanisms in astrocyte-based models, supporting its relevance as a multitarget preclinical candidate for early stage AD mechanisms.

Reproduced under the paper's license (CC BY), from the paper cited above.

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Version 1, 30 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 19 authors, 5 keywords, 15 MeSH terms, 94 references.

Cite

This paper

Hacımüftüoğlu, A., Saraçoğlu, N., Saffour, S., Abad, N., Kesgun, Y., Zegheb, N., Gundeger, E., Yeşilyurt, F., Ataş, M. N., Bati-Ayaz, G., Altunlu, Ö., Çınar, B., Yörük, M. A., Okkay, U., Özkaraca, M., Ateş, O., Taghizadehghalehjoughi, A., Lafzi, F., & Türkez, H. (2026). Multi-Target Neuroprotective Compound Exhibits EAAT2-Modulating and Alzheimer's Pathology-Attenuating Effects in In Vitro and In Vivo Models. ACS chemical neuroscience, 17(10), 1919-1944. https://doi.org/10.1021/acschemneuro.5c00873

BibTeX

@article{hacmuftuoglu2026multi,
author = {Hacımüftüoğlu, Ahmet and Saraçoğlu, Nurullah and Saffour, Sana and Abad, Nadeem and Kesgun, Yunus and Zegheb, Nadjiba and Gundeger, Ersin and Yeşilyurt, Fatma and Ataş, Merve Nur and Bati-Ayaz, Gizem and Altunlu, Öznur and Çınar, Burak and Yörük, Mehmet Ali and Okkay, Ufuk and Özkaraca, Mustafa and Ateş, Orhan and Taghizadehghalehjoughi, Ali and Lafzi, Ferruh and Türkez, Hasan},
title = {{Multi-Target Neuroprotective Compound Exhibits EAAT2-Modulating and Alzheimer's Pathology-Attenuating Effects in In Vitro and In Vivo Models}},
journal = {ACS chemical neuroscience},
year = {2026},
month = apr,
volume = {17},
number = {10},
pages = {1919--1944},
publisher = {American Chemical Society},
issn = {1948-7193},
doi = {10.1021/acschemneuro.5c00873},
url = {https://doi.org/10.1021/acschemneuro.5c00873},
pmid = {42051019},
pmcid = {PMC13195668}
}

RIS

TY - JOUR
AU - Hacımüftüoğlu, Ahmet
AU - Saraçoğlu, Nurullah
AU - Saffour, Sana
AU - Abad, Nadeem
AU - Kesgun, Yunus
AU - Zegheb, Nadjiba
AU - Gundeger, Ersin
AU - Yeşilyurt, Fatma
AU - Ataş, Merve Nur
AU - Bati-Ayaz, Gizem
AU - Altunlu, Öznur
AU - Çınar, Burak
AU - Yörük, Mehmet Ali
AU - Okkay, Ufuk
AU - Özkaraca, Mustafa
AU - Ateş, Orhan
AU - Taghizadehghalehjoughi, Ali
AU - Lafzi, Ferruh
AU - Türkez, Hasan
TI - Multi-Target Neuroprotective Compound Exhibits EAAT2-Modulating and Alzheimer's Pathology-Attenuating Effects in In Vitro and In Vivo Models
T2 - ACS chemical neuroscience
J2 - ACS Chem Neurosci
PY - 2026
DA - 2026/04/29
VL - 17
IS - 10
SP - 1919
EP - 1944
SN - 1948-7193
PB - American Chemical Society
DO - 10.1021/acschemneuro.5c00873
UR - https://doi.org/10.1021/acschemneuro.5c00873
LA - en
ER -

CSL-JSON

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