Fatty acid amide hydrolase inhibitors. 3: Tetra-substituted azetidine ureas with in vivo activity

Roughley, Stephen D.; Browne, Helen; Macías, A.; Benwell, Karen; Brooks, Teresa; D’Alessandro, Jalanie; Daniels, Zoe; Dugdale, Sarah; Francis, Geraint L.; Gibbons, Ben; Hart, Terance W.; Haymes, Timothy; Kennett, Guy A.; Lightowler, S.; Matassova, Natalia; Mansell, Howard L.; Merrett, Angela; Misra, Anil Kumar; Padfield, Anthony; Parsons, Rachel; Pratt, Robert M.; Robertson, Alan; Simmonite, Heather; Tan, Kiri; Walls, Steven B.; Wong, Melanie

doi:10.1016/j.bmcl.2011.12.032

Cited by 10 publications

(9 citation statements)

References 31 publications

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“…In substrate-dependence experiments, that were performed including several data points in the low substrate concentration range, enzymatic activity of rFAAH and its mutants was assayed by means of the fluorescent-based method by Ramarao and colleagues 44 , by measuring the release of arachidonic acid and of the highly fluorescent 7-ammino-4-methyl coumarin (AMC) product from the non-fluorescent arachidonyl 7-ammino-4-methyl coumarin amide (AAMCA) substrate of FAAH. In preliminary experiments both methods (radiometric and fluorimetric) were found to yield the same catalytic constants, that were in keeping with literature data 15,34,45 . Kinetic analysis were done both in the presence and in the absence of 1-palmitoyl-2-oleyl-sn-glycero-3-phosphocholine (POPC) large unilamellar vesicles prepared as described previously 46 .…”

Section: Methodssupporting

confidence: 87%

The endocannabinoid hydrolase FAAH is an allosteric enzyme

Dainese

Oddi

Simonetti

et al. 2020

Sci Rep

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Fatty acid amide hydrolase (FAAH) is a membrane-bound homodimeric enzyme that in vivo controls content and biological activity of N-arachidonoylethanolamine (AEA) and other relevant bioactive lipids termed endocannabinoids. Parallel orientation of FAAH monomers likely allows both subunits to simultaneously recruit and cleave substrates. Here, we show full inhibition of human and rat FAAH by means of enzyme inhibitors used at a homodimer:inhibitor stoichiometric ratio of 1:1, implying that occupation of only one of the two active sites of FAAH is enough to fully block catalysis. Single W445Y substitution in rat FAAH displayed the same activity as the wild-type, but failed to show full inhibition at the homodimer:inhibitor 1:1 ratio. Instead, F432A mutant exhibited reduced specific activity but was fully inhibited at the homodimer:inhibitor 1:1 ratio. Kinetic analysis of AEA hydrolysis by rat FAAH and its F432A mutant demonstrated a Hill coefficient of ~1.6, that instead was ~1.0 in the W445Y mutant. Of note, also human FAAH catalysed an allosteric hydrolysis of AEA, showing a Hill coefficient of ~1.9. Taken together, this study demonstrates an unprecedented allosterism of FAAH, and represents a case of communication between two enzyme subunits seemingly controlled by a single amino acid (W445) at the dimer interface. In the light of extensive attempts and subsequent failures over the last decade to develop effective drugs for human therapy, these findings pave the way to the rationale design of new molecules that, by acting as positive or negative heterotropic effectors of FAAH, may control more efficiently its activity. Endocannabinoids form a relevant class of lipids that are widespread in tissues, where they exert diverse biological actions 1. Endocannabinoids have been demonstrated to reduce pain and inflammation, modulate energetic homeostasis and appetite, and have anticancer, anxiolytic, and neuroprotective effects through type-1 and/or type-2 cannabinoid receptors (CB 1 and CB 2), as well as via stimulation of transient receptor potential vanilloid channels, peroxisome proliferator-activated receptors, and additional targets 2-4. Their degradation is due to the activity of several metabolic enzymes, among which FAAH has a prominent role also in vivo 5. In addition, biotransformation of endocannabinoids is catalysed by lipoxygenases (LOXs) or cyclooxygenase-2 (COX-2) 6-8. FAAH is a membrane-bound homodimer that is able to hydrolyse AEA and, to a lesser extent, 2-arachidonoylglycerol (2-AG), N-oleoylethanolamine 9,10 , N-palmitoylethanolamine 11 , and N-oleoyltaurine 5,12,13. The biological activity of AEA, 2-AG and related compounds within the central nervous system 2,3 and at the periphery has been recently reviewed 6. The 3D structures of a truncated form of rat FAAH lacking the N-terminal region 14 (hereafter referred to as rFAAH), and of its humanized form containing residues of the human active site 15 , have been resolved at high resolution, and some details of the catalytic mechanism of the enzyme ha...

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Section: Methodssupporting

confidence: 87%

The endocannabinoid hydrolase FAAH is an allosteric enzyme

Dainese

Oddi

Simonetti

et al. 2020

Sci Rep

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“…: 2-Chloropyridin-3-yl)-phenylmethanol [20] (18.2 mmol, 1 equiv. ), tert-butyldimethylchlorosilane (20.2 mmol, 1.1 equiv.)…”

Section: -Chloro-(3-{[(tert-butyldimethylsilyl)oxy](phenyl)methyl}pymentioning

confidence: 99%

“…5 (C 8,9,10,11,12, 6,17,18,19,20 ), 122.6 (C 2 ), 79.9 (C 4 ), 54.9 (C 13 ) ppm; HRMS (ESI): m/z calcd. for C 20…”

Section: -(4-bromophenyl)-34-dihydro-1h-2-benzopyran-3-one (Pp1i)mentioning

confidence: 99%

Synthetic Strategy Studies for a Concise Access to Functionalized Pyrano[4,3‐b]pyridin‐7‐ones: An Entry to Semi‐Rigid Analogs of Antihistamines

Beghennou

Passador

et al. 2020

Eur J Org Chem

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We report short and efficient syntheses of polyfunctionalized 5,8-dihydro-7H-pyrano[4,3-b]pyridin-7-ones and 1,4dihydro-3H-pyrano[4,3-c]pyridin-3-ones which can be considered as new aza analogs of 3-isochromanones and as promising scaffolds for medicinal chemistry. Depending on the nature of the substituent, three different and complementary synthetic methodologies were used allowing the introduction of significant diversity in the substituent on the lactone ring of the pyr

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“…This compound was reported to be a time-dependent inhibitor of human FAAH, displaying an IC 50 value of 1031 nM, after 1 h of pre-incubation. Vernalis attempted to improve the in vitro potency on human FAAH, as well as to reduce microsomal metabolism of azetidin-1-yl(piperidin-1-yl)methanone analogs of VER-156084, but without success interrupted the development of VER-156084 in favor of alternative chemotypes [81]. The second series developed by Vernalis was characterized by an N-(pyridazin-3-yl)azetidine-1-carboxamide core, as exemplified by compound 43, which inhibits human FAAH with an IC 50 of 38 nM after 1 h of pre-incubation.…”

Section: Aryl Ureasmentioning

confidence: 99%

“…This class of compounds was identified through an HTS campaign on human FAAH using a microsomal protein preparation and a fluorescent assay readout [109]. The first hit (the tetrahydropyridopyrimidine 81) was the subject of an intensive optimization program that led to the tetrahydronaphthyridine 82 (RN-450), able to inhibit human and rat FAAH with IC 50 values of 13 and 25 nM, respectively. The mechanism of action of RN-450 was thoroughly investigated and it was shown that this compound acts as a competitive and reversible FAAH inhibitor.…”

Section: Tetrahydronaphthyridine Derivativesmentioning

confidence: 99%

Fatty acid amide hydrolase inhibitors: a patent review (2009 – 2014)

Lodola

Castelli

Mor

et al. 2015

Expert Opinion on Therapeutic Patents

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FAAH is a promising target for treating many disease conditions including pain, inflammation and mood disorders. In the last few years, remarkable efforts have been made to develop new FAAH inhibitors (either reversible and irreversible) characterized by excellent potency and selectivity, to complete the arsenal of tools for modulating FAAH activity. The failure of PF-04457845 in a Phase II study on osteoarthritis pain has not flattened the interest in FAAH inhibitors. New clinical trials on 'classical' FAAH inhibitors are now ongoing, and new strategies based on compounds with peculiar in vivo distribution (e.g., peripheral) or with multiple pharmacological activities (e.g., FAAH and COX) are under investigation and could boost the therapeutic potential of this class in the next future.

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Fatty acid amide hydrolase inhibitors. 3: Tetra-substituted azetidine ureas with in vivo activity

Cited by 10 publications

References 31 publications

The endocannabinoid hydrolase FAAH is an allosteric enzyme

The endocannabinoid hydrolase FAAH is an allosteric enzyme

Synthetic Strategy Studies for a Concise Access to Functionalized Pyrano[4,3‐b]pyridin‐7‐ones: An Entry to Semi‐Rigid Analogs of Antihistamines

Fatty acid amide hydrolase inhibitors: a patent review (2009 – 2014)

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