Fatty acid amide hydrolase inhibitors. 2. Novel synthesis of sterically hindered azabenzhydryl ethers and an improved synthesis of VER-156084

We report an improved synthesis of the fatty acid amide hydrolase (FAAH) inhibitor VER-156084. The key step is a novel, environmentally benign etherification to form an unusual, highly hindered azabenzhydryl ether. The method is applied to a variety of primary and secondary alcohols.     

脑胶质瘤中脂肪酸酰胺水解酶的表达及意义

目的研究内源性大麻素代谢酶脂肪酸酰胺水解酶(FAAH)在脑胶质瘤组织中的表达情况及其与胶质瘤病理级别的关系,探讨其临床意义.方法选取首次手术切除并经病理检查证实的脑胶质瘤标本23例,其中WHOⅠ级1例,Ⅱ级7例,Ⅲ级5例,Ⅲ~Ⅳ级2例,Ⅳ级8例.另取8例因颅脑外伤行内减压术切除的正常脑组织作为对照.应用实时荧光定量PCR和高效液相色谱-质谱联用技术分别检测各组织标本中FAAH mRNA和FAAH活性的表达.结果 FAAH mRNA、FAAH活性在低级别(WHOⅠ~Ⅱ级)和高级别(WHOⅢ~Ⅳ级)脑胶质瘤组织中的表达水平较正常脑组织明显降低,差异有统计学意义(均P<0.01),且随着肿瘤病理级别的增高而降低,与肿瘤的病理分级呈负相关(均P<0.01).FAAH mRNA与FAAH活性的表达水平呈正相关(P<0.01).结论 FAAH可能参与了脑胶质瘤的发生、发展过程,检测FAAH的表达可评价脑胶质瘤的生物学行为,为脑胶质瘤的临床治疗和预后判断提供参考.     

In Silico and In Vitro Analysis of Major Cannabis-Derived Compounds as Fatty Acid Amide Hydrolase Inhibitors

Accumulated evidence suggests that enhancing the endocannabinoid (eCB) tone, in particular of anandamide (N-arachidonoylethanolamine, AEA), has therapeutic potential in many human diseases. Fatty acid amide hydrolase (FAAH) is a membrane-bound enzyme principally responsible for the degradation of AEA, and thus it represents a relevant target to increase signaling thereof. In recent years, different synthetic and natural compounds have been developed and tested on rat FAAH, but little is known of their effect on the human enzyme. Here, we sought to investigate six major cannabis-derived compounds to compare their action on rat and human FAAHs. To this aim, we combined an in silico analysis of their binding mode and affinity, with in vitro assays of their effect on enzyme activity. This integrated approach allowed to disclose differences in efficacy towards rat and human FAAHs, and to highlight the role of key residues involved in the inhibition of both enzymes. This study suggests that the therapeutic efficacy of compounds targeted towards FAAH should be always tested in vitro on both rat and human enzymes.     

New Coumarin Derivatives as Cholinergic and Cannabinoid System Modulators

In the last years, the connection between the endocannabinoid system (eCS) and neuroprotection has been discovered, and evidence indicates that eCS signaling is involved in the regulation of cognitive processes and in the pathophysiology of Alzheimer’s disease (AD). Accordingly, pharmacotherapy targeting eCS could represent a valuable contribution in fighting a multifaceted disease such as AD, opening a new perspective for the development of active agents with multitarget potential. In this paper, a series of coumarin-based carbamic and amide derivatives were designed and synthesized as multipotent compounds acting on cholinergic system and eCS-related targets. Indeed, they were tested with appropriate enzymatic assays on acetyl and butyryl-cholinesterases and on fatty acid amide hydrolase (FAAH), and also evaluated as cannabinoid receptor (CB1 and CB2) ligands. Moreover, their ability to reduce the self-aggregation of beta amyloid protein (Aβ₄₂) was assessed. Compounds 2 and 3, bearing a carbamate function, emerged as promising inhibitors of hAChE, hBuChE, FAAH and Aβ₄₂ self-aggregation, albeit with moderate potencies, while the amide 6 also appears a promising CB1/CB2 receptors ligand. These data prove for the new compounds an encouraging multitarget profile, deserving further evaluation.     

Microwave-assisted four-component reaction for the synthesis of a monothiohydantoin inhibitor of a fatty acid amide hydrolase

Monothiohydantoin 3a is made in seven steps from 1,4-diiodobenzene and is shown to be an inhibitor of fatty acid amide hydrolase (IC₅₀ = 23.4 ± 1.1 μM). The key step in the sequence involves a microwave-assisted four-component reaction that makes the 5,5′-disubstituted hydantoin nucleus by the sequential formation of two C-C and two C-N bonds.     

Tandem mass spectrometric data-FAAH inhibitory activity relationships of some carbamic acid O-aryl esters

We have recently described a class of systemically active inhibitors of the intracellular activity of fatty acid amide hydrolase (FAAH) and traced extensive structure-activity relationships. These compounds, characterized by an N-alkyl carbamic acid O-aryl ester structure, exert potent anxiolytic-like effects in animal models. In the present study, possible relationships between mass spectrometric parameters (related to the propensity of the C(O)--O bond to be cleaved) and FAAH-inhibitory potency were tested. With this aim, a set of our products was analyzed by electrospray ionization mass spectrometry and the protonated molecules were decomposed by low-energy collisions. The experiments were performed by ion trap mass spectrometry, which led to a step-by-step energy deposition, thus favouring the lowest critical energy decomposition channels. For all compounds, breakdown curves relative to [MH](+) ions and to the fragment implying C(O)--O bond cleavage were obtained. The crossing point between these curves was related to the energetics of decomposition and the values found for the investigated compounds were linearly correlated (r(2) = 0.797) with their FAAH-inhibitory activity. This indicates that the energetics of the C(O)--O bond cleavage may be relevant in explaining FAAH inhibition.