BACE Biological Assays

Tomasselli, Alfredo G.; Bienkowski, Michael J.

doi:10.1002/9780470594087.ch3

Cited by 3 publications

(2 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since the final protein concentrations in this study exceed those used by Qian et al [ 34 ] by fivefold (20 vs. 100 μg/mL), it is possible that there is residual metabolic activity associated with the higher protein. However, K i is a measurement of affinity that is independent of protein concentration [ 42 ]. Thus, it is also possible that CES1 has multiple catalytic sites which are substrate-specific.…”

Section: Discussionmentioning

confidence: 99%

Characterisation of AMB-FUBINACA metabolism and CB1-mediated activity of its acid metabolite

et al. 2022

View full text Add to dashboard Cite

Purpose AMB-FUBINACA is a synthetic cannabinoid receptor agonist (SCRA) which is primarily metabolised by hepatic enzymes producing AMB-FUBINACA carboxylic acid. The metabolising enzymes associated with this biotransformation remain unknown. This study aimed to determine if AMB-FUBINACA metabolism could be reduced in the presence of carboxylesterase (CES) inhibitors and recreational drugs commonly consumed with it. The affinity and activity of the AMB-FUBINACA acid metabolite at the cannabinoid type-1 receptor (CB1) was investigated to determine the activity of the metabolite. Methods The effect of CES1 and CES2 inhibitors, and delta-9-tetrahydrocannabinol (Δ9-THC) on AMB-FUBINACA metabolism were determined using both human liver microsomes (HLM) and recombinant carboxylesterases. Radioligand binding and cAMP assays comparing AMB-FUBINACA and AMB-FUBINACA acid were carried out in HEK293 cells expressing human CB1. Results AMB-FUBINACA was rapidly metabolised by HLM in the presence and absence of NADPH. Additionally, CES1 and CES2 inhibitors both significantly reduced AMB-FUBINACA metabolism. Furthermore, digitonin (100 µM) significantly inhibited CES1-mediated metabolism of AMB-FUBINACA by ~ 56%, while the effects elicited by Δ9-THC were not statistically significant. AMB-FUBINACA acid produced only 26% radioligand displacement consistent with low affinity binding. In cAMP assays, the potency of AMB-FUBINACA was ~ 3000-fold greater at CB1 as compared to the acid metabolite. Conclusions CES1A1 was identified as the main hepatic enzyme responsible for the metabolism of AMB-FUBINACA to its less potent carboxylic acid metabolite. This biotransformation was significantly inhibited by digitonin. Since other xenobiotics may also inhibit similar SCRA metabolic pathways, understanding these interactions may elucidate why some users experience high levels of harm following SCRA use.

show abstract

Section: Discussionmentioning

confidence: 99%

Characterisation of AMB-FUBINACA metabolism and CB1-mediated activity of its acid metabolite

et al. 2022

View full text Add to dashboard Cite

show abstract

“…A catalytic water molecule lies between the two acidic residues, becoming activated by abstraction of a proton in the pH range of 4.5–5.0. This activated water then serves as the nucleophile which cleaves the scissile peptide bond in a substrate polypeptide. ,, Sequence specificity in the substrate polypeptide is achieved through recognition of side chain residues in the substrate via specificity pockets on the N-terminal side (Sn) and C-terminal side (Sn′) of the scissile peptide bond. − …”

Section: Introductionmentioning

confidence: 99%

Discovery and Optimization of a Novel Spiropyrrolidine Inhibitor of β-Secretase (BACE1) through Fragment-Based Drug Design

et al. 2012

View full text Add to dashboard Cite

The aspartyl protease β-secretase, or BACE, has been demonstrated to be a key factor in the proteolytic formation of Aβ-peptide, a major component of plaques in the brains of Alzheimer's disease (AD) patients, and inhibition of this enzyme has emerged as a major strategy for pharmacologic intervention in AD. An X-ray-based fragment screen of Pfizer's proprietary fragment collection has resulted in the identification of a novel BACE binder featuring spiropyrrolidine framework. Although exhibiting only weak inhibitory activity against the BACE enzyme, the small compound was verified by biophysical and NMR-based methods as a bona fide BACE inhibitor. Subsequent optimization of the lead compound, relying heavily on structure-based drug design and computational prediction of physiochemical properties, resulted in a nearly 1000-fold improvement in potency while maintaining ligand efficiency and properties predictive of good permeability and low P-gp liability.

show abstract

Identification of new BACE1 inhibitors using Pharmacophore and Molecular dynamics simulations approach

Dhanabalan

Kesherwani

Velmurugan

et al. 2017

Journal of Molecular Graphics and Modelling

View full text Add to dashboard Cite

BACE Biological Assays

Cited by 3 publications

References 41 publications

Characterisation of AMB-FUBINACA metabolism and CB1-mediated activity of its acid metabolite

Characterisation of AMB-FUBINACA metabolism and CB1-mediated activity of its acid metabolite

Discovery and Optimization of a Novel Spiropyrrolidine Inhibitor of β-Secretase (BACE1) through Fragment-Based Drug Design

Identification of new BACE1 inhibitors using Pharmacophore and Molecular dynamics simulations approach

Contact Info

Product

Resources

About