Computational and Experimental Evidence for the Neuroprotective Role of 4-Methyl-2-(5-phenyl-6-thioxo-1,3,5-thiadiazinan-3-yl)pentanoic Acid via NRF2/TLR4 Signaling in Aging Mice

Saleha Gul; Gul Nabi Khan; Shahid Ali Shah; Rasool Khan; Haleema Ali; Umer Sadique; Farrah Zaidi

doi:10.70749/ijbr.v3i8.2162

Authors

Saleha Gul Institute of Zoological Sciences, University of Peshawar, Pakistan. Department of Zoology, Islamia College University, Peshawar, Pakistan https://orcid.org/0009-0004-5031-2130
Gul Nabi Khan Department of Zoology, Islamia College University, Peshawar, Pakistan https://orcid.org/0000-0002-8740-9815
Shahid Ali Shah Department of Biology, University of Haripur, Haripur, Khyber Pakhtunkhwa, Pakistan https://orcid.org/0000-0002-5341-646X
Rasool Khan Institute of Chemical Sciences, University of Peshawar, Pakistan https://orcid.org/0009-0009-6618-5849
Haleema Ali Institute of Chemical Sciences, University of Peshawar, Pakistan https://orcid.org/0000-0002-8163-0017
Umer Sadique College of Veterinary Sciences, Faculty of Animal Husbandry and Veterinary Sciences, University of Agriculture, Peshawar, Pakistan https://orcid.org/0009-0007-5354-9063
Farrah Zaidi Institute of Zoological Sciences, University of Peshawar, Pakistan https://orcid.org/0009-0008-8684-9537

DOI:

https://doi.org/10.70749/ijbr.v3i8.2162

Keywords:

D-Gal, Pentanoic Acid, Mice, TLR4, Nrf-2, Astrocytes, Microglia

Abstract

Background: Aging is an inevitable biological process and a significant risk factor for numerous neurodegenerative and psychiatric diseases. Aim: The current study offers the first complete assessment of the neuroprotective effects of a novel pentanoic acid derivative (PAD), 4-Methyl-2-(5-Phenyl-Thioxo-1,3,5-Thiadiazinan-3-yl), in a D-galactose (D-Gal)-induced model of brain aging in male mice. Material and methods: Thirty males were assigned randomly to six groups (n = 10); a normal control, a D-Gal group (100 mg/kg), and a D-Gal + PAD group (25 mg/kg). D-galactose was given intraperitoneally once a day for 56 days whereas PAD was started at day 29 for 28 days. Results: Western blotting showed that chronic exposure to D-galactose induced microglial and astrocytic activation, increased TLR4 and NF-κB signaling, decreased Nrf2 expression, and increased PARP-1 activity in the mouse brain. An administration of PAD inhibited these changes by reducing glial activation, the level of TLR4, and increasing Nrf2, and reducing the activity of PARP-1. In Silico molecular docking studies were conducted for further mechanistic validation, which demonstrated that PAD forms stronger stable hydrogen bonds with specific residues within the KEAP1 binding pocket that may disrupt the KEAP1–NRF2 complex and initiate NRF2 signaling. Furthermore, PAD showed high binding affinity toward the TLR4 active site, indicating inhibition of TLR4 mediated inflammatory signaling. Conclusion: Taken together, these results demonstrate that this new PAD compound demonstrated neuroprotective effects through dual modulation of NRF2 and TLR4 pathways. The PAD compound represents a promising drug candidate for age-associated neurodegenerative diseases due to its potential to act as both a NRF2 activator and TLR4 inhibitor. Further pharmacologic and mechanistic investigation will be required to substantiate its drug-like features and therapeutic efficacy.

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