BChE-IN-44 is a potent and highly selective butyrylcholinesterase (BChE) inhibitor that can penetrate the blood-brain barrier, demonstrated by [equineBChEIC50= 18.00 pM, humanBChEIC50= 1.50 nM]. BChE-IN-44 offers neuroprotective effects against Aβ1-42-induced damage models and halts the self-aggregation of Aβ1-42. It reduces levels of inflammatory factors (NO, IL-6, and TNF-α) in BV2 cells induced by lipopolysaccharides. Additionally, BChE-IN-44 significantly ameliorates cognitive impairment caused by scopolamine. It also regulates BChE and acetylcholine levels in the hippocampus of mice. BChE-IN-44 is applicable for Alzheimer's disease (AD) research.

BChE-IN-44 (Compound N14) exhibits approximately 15% toxicity towards PC12 and SH-SY5Y cells at 50 μM for 24 hours, indicating its high in vitro safety. The compound has a Kc value of 0.017 μM, demonstrating significant affinity for BChE. At concentrations of 5-20 μM for 24 hours, BChE-IN-44 significantly reduces levels of inflammatory factors (NO, IL-6, and TNF-α) in LPS-induced BV2 cells. Furthermore, at 20 μM for 24 hours, BChE-IN-44 provides substantial neuroprotection against Aβ 1-42-induced damage in PC12 cells. Additionally, at 20 μM for 48 hours, it effectively inhibits Aβ 1-42 aggregation, confirming the carrier scaffold's effective inhibition of Aβ1-42 aggregation.

BChE-IN-44 (5-20 mg/kg, intraperitoneal injection, once daily for 14 days) effectively reverses cognitive impairment, anxiety, and learning and memory deficits induced by Scopolamine in mice. It inhibits BChE activity, increases ACh levels, and reduces neuroinflammation.