Niphakis MJ

References (35)

Title : Attenuating ABHD17 enhances S- palmitoylation, membrane localization and signal transduction of NOD2 and Crohn's disease-associated variants - Dixon_2023_bioRxiv__
Author(s) : Dixon CL , Martin NR , Niphakis MJ , Cravatt BF , Fairn GD
Ref : Biorxiv , : , 2023
Abstract : NOD2 is an intracellular innate immune receptor that senses bacterial peptidoglycans. Although soluble in the cytosol, a portion of the protein is associated with the plasma membrane and endosomal compartments for microbial surveillance. Palmitoylation of NOD2 by zDHHC5 promotes its membrane recruitment to drive proinflammatory and antimicrobial responses to pathogenic invasion. A depalmitoylation step by an unknown protein, thioesterase, releases NOD2 from membranes into the cytosol, where the protein can then enter a new cycle of palmitoylation-depalmitoylation. Here, we identify alpha/beta -hydrolase domain-containing protein 17 isoforms (ABHD17A, 17B, 17C) as the thioesterases responsible for depalmitoylation of NOD2. Inhibiting ABHD17 increased the plasmalemmal localization of both wild-type NOD2 and a subset of hypo-palmitoylated Crohn's disease-associated variants, resulting in increased NF-kappaB activation and production of pro-inflammatory cytokines in epithelial cells. These results suggest that targeted inhibition of ABHD17 may rescue some Crohn's disease-associated NOD2 variants.
ESTHER : Dixon_2023_bioRxiv__
PubMedSearch : Dixon_2023_bioRxiv__
PubMedID: 38187608

Title : Chemoproteomic identification of a DPP4 homolog in Bacteroides thetaiotaomicron - Keller_2023_Nat.Chem.Biol__
Author(s) : Keller LJ , Nguyen TH , Liu LJ , Hurysz BM , Lakemeyer M , Guerra M , Gelsinger DJ , Chanin R , Ngo N , Lum KM , Faucher F , Ipock P , Niphakis MJ , Bhatt AS , O'Donoghue AJ , Huang KC , Bogyo M
Ref : Nat Chemical Biology , : , 2023
Abstract : Serine hydrolases have important roles in signaling and human metabolism, yet little is known about their functions in gut commensal bacteria. Using bioinformatics and chemoproteomics, we identify serine hydrolases in the gut commensal Bacteroides thetaiotaomicron that are specific to the Bacteroidetes phylum. Two are predicted homologs of the human dipeptidyl peptidase 4 (hDPP4), a key enzyme that regulates insulin signaling. Our functional studies reveal that BT4193 is a true homolog of hDPP4 that can be inhibited by FDA-approved type 2 diabetes medications targeting hDPP4, while the other is a misannotated proline-specific triaminopeptidase. We demonstrate that BT4193 is important for envelope integrity and that loss of BT4193 reduces B. thetaiotaomicron fitness during in vitro growth within a diverse community. However, neither function is dependent on BT4193 proteolytic activity, suggesting a scaffolding or signaling function for this bacterial protease.
ESTHER : Keller_2023_Nat.Chem.Biol__
PubMedSearch : Keller_2023_Nat.Chem.Biol__
PubMedID: 37349583

Title : Inhibitory Neurotransmission Is Sex-Dependently Affected by Tat Expression in Transgenic Mice and Suppressed by the Fatty Acid Amide Hydrolase Enzyme Inhibitor PF3845 via Cannabinoid Type-1 Receptor Mechanisms - Xu_2022_Cells_11_
Author(s) : Xu C , Yadav-Samudrala BJ , Nath B , Mistry T , Jiang W , Niphakis MJ , Cravatt BF , Mukhopadhyay S , Lichtman AH , Ignatowska-Jankowska BM , Fitting S
Ref : Cells , 11 : , 2022
Abstract : (1) Background. The endocannabinoid (eCB) system, which regulates physiological and cognitive processes, presents a promising therapeutic target for treating HIV-associated neurocognitive disorders (HAND). Here we examine whether upregulating eCB tone has potential protective effects against HIV-1 Tat (a key HIV transactivator of transcription) protein-induced alterations in synaptic activity. (2) Methods. Whole-cell patch-clamp recordings were performed to assess inhibitory GABAergic neurotransmission in prefrontal cortex slices of Tat transgenic male and female mice, in the presence and absence of the fatty acid amide hydrolase (FAAH) enzyme inhibitor PF3845. Western blot and mass spectrometry analyses assessed alterations of cannabinoid receptor and enzyme protein expression as well as endogenous ligands, respectively, to determine the impact of Tat exposure on the eCB system. (3) Results. GABAergic activity was significantly altered upon Tat exposure based on sex, whereas the effectiveness of PF3845 to suppress GABAergic activity in Tat transgenic mice was not altered by Tat or sex and involved CB(1)R-related mechanisms that depended on calcium signaling. Additionally, our data indicated sex-dependent changes for AEA and related non-eCB lipids based on Tat induction. (4) Conclusion. Results highlight sex- and/or Tat-dependent alterations of GABAergic activity and eCB signaling in the prefrontal cortex of Tat transgenic mice and further increase our understanding about the role of FAAH inhibition in neuroHIV.
ESTHER : Xu_2022_Cells_11_
PubMedSearch : Xu_2022_Cells_11_
PubMedID: 35269478

Title : Monoacylglycerol Lipase Inhibitor MJN110 Reduces Neuronal Hyperexcitability, Restores Dendritic Arborization Complexity, and Regulates Reward-Related Behavior in Presence of HIV-1 Tat - League_2021_Front.Neurol_12_651272
Author(s) : League AF , Gorman BL , Hermes DJ , Johnson CT , Jacobs IR , Yadav-Samudrala BJ , Poklis JL , Niphakis MJ , Cravatt BF , Lichtman AH , Ignatowska-Jankowska BM , Fitting S
Ref : Front Neurol , 12 :651272 , 2021
Abstract : While current therapeutic strategies for people living with human immunodeficiency virus type 1 (HIV-1) suppress virus replication peripherally, viral proteins such as transactivator of transcription (Tat) enter the central nervous system early upon infection and contribute to chronic inflammatory conditions even alongside antiretroviral treatment. As demand grows for supplemental strategies to combat virus-associated pathology presenting frequently as HIV-associated neurocognitive disorders (HAND), the present study aimed to characterize the potential utility of inhibiting monoacylglycerol lipase (MAGL) activity to increase inhibitory activity at cannabinoid receptor-type 1 receptors through upregulation of 2-arachidonoylglycerol (2-AG) and downregulation of its degradation into proinflammatory metabolite arachidonic acid (AA). The MAGL inhibitor MJN110 significantly reduced intracellular calcium and increased dendritic branching complexity in Tat-treated primary frontal cortex neuron cultures. Chronic MJN110 administration in vivo increased 2-AG levels in the prefrontal cortex (PFC) and striatum across Tat(+) and Tat(-) groups and restored PFC N-arachidonoylethanolamine (AEA) levels in Tat(+) subjects. While Tat expression significantly increased rate of reward-related behavioral task acquisition in a novel discriminative stimulus learning and cognitive flexibility assay, MJN110 altered reversal acquisition specifically in Tat(+) mice to rates indistinguishable from Tat(-) controls. Collectively, our results suggest a neuroprotective role of MAGL inhibition in reducing neuronal hyperexcitability, restoring dendritic arborization complexity, and mitigating neurocognitive alterations driven by viral proteins associated with latent HIV-1 infection.
ESTHER : League_2021_Front.Neurol_12_651272
PubMedSearch : League_2021_Front.Neurol_12_651272
PubMedID: 34484091
Gene_locus related to this paper: human-MGLL

Title : ABHD17 regulation of plasma membrane palmitoylation and N-Ras-dependent cancer growth - Remsberg_2021_Nat.Chem.Biol__
Author(s) : Remsberg JR , Suciu RM , Zambetti NA , Hanigan TW , Firestone AJ , Inguva A , Long A , Ngo N , Lum KM , Henry CL , Richardson SK , Predovic M , Huang B , Dix MM , Howell AR , Niphakis MJ , Shannon K , Cravatt BF
Ref : Nat Chemical Biology , : , 2021
Abstract : Multiple Ras proteins, including N-Ras, depend on a palmitoylation/depalmitoylation cycle to regulate their subcellular trafficking and oncogenicity. General lipase inhibitors such as Palmostatin M (Palm M) block N-Ras depalmitoylation, but lack specificity and target several enzymes displaying depalmitoylase activity. Here, we describe ABD957, a potent and selective covalent inhibitor of the ABHD17 family of depalmitoylases, and show that this compound impairs N-Ras depalmitoylation in human acute myeloid leukemia (AML) cells. ABD957 produced partial effects on N-Ras palmitoylation compared with Palm M, but was much more selective across the proteome, reflecting a plasma membrane-delineated action on dynamically palmitoylated proteins. Finally, ABD957 impaired N-Ras signaling and the growth of NRAS-mutant AML cells in a manner that synergizes with MAP kinase kinase (MEK) inhibition. Our findings uncover a surprisingly restricted role for ABHD17 enzymes as regulators of the N-Ras palmitoylation cycle and suggest that ABHD17 inhibitors may have value as targeted therapies for NRAS-mutant cancers.
ESTHER : Remsberg_2021_Nat.Chem.Biol__
PubMedSearch : Remsberg_2021_Nat.Chem.Biol__
PubMedID: 33927411
Gene_locus related to this paper: human-ABHD17A , human-ABHD17B , human-ABHD17C

Title : Toxoplasma gondii serine hydrolases regulate parasite lipid mobilization during growth and replication within the host - Onguka_2021_Cell.Chem.Biol__
Author(s) : Onguka O , Babin BM , Lakemeyer M , Foe IT , Amara N , Terrell SM , Lum KM , Cieplak P , Niphakis MJ , Long JZ , Bogyo M
Ref : Cell Chemical Biology , : , 2021
Abstract : The intracellular protozoan parasite Toxoplasma gondii must scavenge cholesterol and other lipids from the host to facilitate intracellular growth and replication. Enzymes responsible for neutral lipid synthesis have been identified but there is no evidence for enzymes that catalyze lipolysis of cholesterol esters and esterified lipids. Here, we characterize several T. gondii serine hydrolases with esterase and thioesterase activities that were previously thought to be depalmitoylating enzymes. We find they do not cleave palmitoyl thiol esters but rather hydrolyze short-chain lipid esters. Deletion of one of the hydrolases results in alterations in levels of multiple lipids species. We also identify small-molecule inhibitors of these hydrolases and show that treatment of parasites results in phenotypic defects reminiscent of parasites exposed to excess cholesterol or oleic acid. Together, these data characterize enzymes necessary for processing lipids critical for infection and highlight the potential for targeting parasite hydrolases for therapeutic applications.
ESTHER : Onguka_2021_Cell.Chem.Biol__
PubMedSearch : Onguka_2021_Cell.Chem.Biol__
PubMedID: 34043961
Gene_locus related to this paper: toxgo-TgASH3 , toxgo-TgASH2 , toxgo-s4ug63 , toxgo-a0a125yz56

Title : AIG1 and ADTRP are endogenous hydrolases of fatty acid esters of hydroxy fatty acids (FAHFAs) in mice - Erikci_2020_J.Biol.Chem_295_5891
Author(s) : Erikci Ertunc M , Kok BP , Parsons WH , Wang JG , Tan D , Donaldson CJ , Pinto AFM , Vaughan JM , Ngo N , Lum KM , Henry CL , Coppola AR , Niphakis MJ , Cravatt BF , Saez E , Saghatelian A
Ref : Journal of Biological Chemistry , 295 :5891 , 2020
Abstract : Fatty acid esters of hydroxy fatty acids (FAHFAs) are a newly discovered class of signaling lipids with anti-inflammatory and anti-diabetic properties. However, the endogenous regulation of FAHFAs remains a pressing but unanswered question. Here, using MS-based FAHFA hydrolysis assays, LC-MS-based lipidomics analyses, and activity-based protein profiling, we found that androgen-induced gene 1 (AIG1) and androgen-dependent TFPI-regulating protein (ADTRP), two threonine hydrolases, control FAHFA levels in vivo in both genetic and pharmacologic mouse models. Tissues from mice lacking ADTRP (Adtrp-KO), or both AIG1 and ADTRP (DKO) had higher concentrations of FAHFAs particularly isomers with the ester bond at the 9(th) carbon due to decreased FAHFA hydrolysis activity. The levels of other lipid classes were unaltered indicating that AIG1 and ADTRP specifically hydrolyze FAHFAs. Complementing these genetic studies, we also identified a dual AIG1/ADTRP inhibitor, ABD-110207, which is active in vivo Acute treatment of WT mice with ABD-110207 resulted in elevated FAHFA levels, further supporting the notion that AIG1 and ADTRP activity control endogenous FAHFA levels. However, loss of AIG1/ADTRP did not mimic the changes associated with pharmacologically administered FAHFAs on extent of upregulation of FAHFA levels, glucose tolerance, or insulin sensitivity in mice, indicating that therapeutic strategies should weigh more on FAHFA administration. Together, these findings identify AIG1 and ADTRP as the first endogenous FAHFA hydrolases identified and provide critical genetic and chemical tools for further characterization of these enzymes and endogenous FAHFAs to unravel their physiological functions and roles in health and disease.
ESTHER : Erikci_2020_J.Biol.Chem_295_5891
PubMedSearch : Erikci_2020_J.Biol.Chem_295_5891
PubMedID: 32152231

Title : The Endocannabinoid System Alleviates Pain in a Murine Model of Cancer-Induced Bone Pain - Thompson_2020_J.Pharmacol.Exp.Ther_373_230
Author(s) : Thompson AL , Grenald SA , Ciccone HA , BassiriRad N , Niphakis MJ , Cravatt BF , Largent-Milnes TM , Vanderah TW
Ref : Journal of Pharmacology & Experimental Therapeutics , 373 :230 , 2020
Abstract : Metastatic breast cancer is prevalent worldwide, and one of the most common sites of metastasis is long bones. Of patients with disease, the major symptom is pain, yet current medications fail to adequately result in analgesic efficacy and present major undesirable adverse effects. In our study, we investigate the potential of a novel monoacylglycerol lipase (MAGL) inhibitor, MJN110, in a murine model of cancer-induced bone pain. Literature has previously demonstrated that MAGL inhibitors function to increase the endogenous concentrations of 2-arachydonylglycerol, which then activates CB1 and CB2 receptors to inhibit inflammation and pain. We demonstrate that administration of MJN110 significantly and dose dependently alleviates spontaneous pain behavior during acute administration compared with vehicle control. In addition, MJN110 maintains its efficacy in a chronic-dosing paradigm over the course of 7 days without signs of receptor sensitization. In vitro analysis of MJN110 demonstrated a dose-dependent and significant decrease in cell viability and proliferation of 66.1 breast adenocarcinoma cells to a greater extent than KML29, an alternate MAGL inhibitor, or the CB2 agonist JWH015. Chronic administration of the compound did not appear to affect tumor burden, as evidenced by radiograph or histologic analysis. Together, these data support the application for MJN110 as a novel therapeutic for cancer-induced bone pain. SIGNIFICANCE STATEMENT: Current standard of care for metastatic breast cancer pain is opioid-based therapies with adjunctive chemotherapy, which have highly addictive and other deleterious side effects. The need for effective, non-opioid-based therapies is essential, and harnessing the endogenous cannabinoid system is proving to be a new target to treat various types of pain conditions. We present a novel drug targeting the endogenous cannabinoid system that is effective at reducing pain in a mouse model of metastatic breast cancer to bone.
ESTHER : Thompson_2020_J.Pharmacol.Exp.Ther_373_230
PubMedSearch : Thompson_2020_J.Pharmacol.Exp.Ther_373_230
PubMedID: 32054717
Gene_locus related to this paper: human-MGLL

Title : Single-Cell Profiling and SCOPE-Seq Reveal Lineage Dynamics of Adult Ventricular-Subventricular Zone Neurogenesis and NOTUM as a Key Regulator - Mizrak_2020_Cell.Rep_31_107805
Author(s) : Mizrak D , Bayin NS , Yuan J , Liu Z , Suciu RM , Niphakis MJ , Ngo N , Lum KM , Cravatt BF , Joyner AL , Sims PA
Ref : Cell Rep , 31 :107805 , 2020
Abstract : In the adult ventricular-subventricular zone (V-SVZ), neural stem cells (NSCs) generate new olfactory bulb (OB) neurons and glia throughout life. To map adult neuronal lineage progression, we profiled >56,000 V-SVZ and OB cells by single-cell RNA sequencing (scRNA-seq). Our analyses reveal the molecular diversity of OB neurons, including fate-mapped neurons, lineage progression dynamics, and an NSC intermediate enriched for Notum, which encodes a secreted WNT antagonist. SCOPE-seq technology, which links live-cell imaging with scRNA-seq, uncovers cell-size transitions during NSC differentiation and preferential NOTUM binding to proliferating neuronal precursors. Consistently, application of NOTUM protein in slice cultures and pharmacological inhibition of NOTUM in slice cultures and in vivo demonstrated that NOTUM negatively regulates V-SVZ proliferation. Timely, context-dependent neurogenesis demands adaptive signaling among neighboring progenitors. Our findings highlight a critical regulatory state during NSC activation marked by NOTUM, which attenuates WNT-stimulated proliferation in NSC progeny.
ESTHER : Mizrak_2020_Cell.Rep_31_107805
PubMedSearch : Mizrak_2020_Cell.Rep_31_107805
PubMedID: 32579931
Gene_locus related to this paper: human-NOTUM

Title : Monoacylglycerol Lipase Inhibitors Reverse Paclitaxel-Induced Nociceptive Behavior and Proinflammatory Markers in a Mouse Model of Chemotherapy-Induced Neuropathy - Curry_2018_J.Pharmacol.Exp.Ther_366_169
Author(s) : Curry ZA , Wilkerson JL , Bagdas D , Kyte SL , Patel N , Donvito G , Mustafa MA , Poklis JL , Niphakis MJ , Hsu KL , Cravatt BF , Gewirtz DA , Damaj MI , Lichtman AH
Ref : Journal of Pharmacology & Experimental Therapeutics , 366 :169 , 2018
Abstract : Although paclitaxel effectively treats various cancers, its debilitating peripheral neuropathic pain side effects often persist long after treatment has ended. Therefore, a compelling need exists for the identification of novel pharmacologic strategies to mitigate this condition. As inhibitors of monoacylglycerol lipase (MAGL), the primary hydrolytic enzyme of the endogenous cannabinoid, 2-arachidonyolglycerol, produces antinociceptive effects in numerous rodent models of pain, we investigated whether inhibitors of this enzyme (i.e., JZL184 and MJN110) would reverse paclitaxel-induced mechanical allodynia in mice. These drugs dose dependently reversed allodynia with respective ED(50) values (95% confidence limit) of 8.4 (5.2-13.6) and 1.8 (1.0-3.3) mg/kg. Complementary genetic and pharmacologic approaches revealed that the antiallodynic effects of each drug require both cannabinoid receptors, CB(1) and CB(2) MJN110 reduced paclitaxel-mediated increased expression of monocyte chemoattractant protein-1 (MCP-1, CCL2) and phospho-p38 MAPK in dorsal root ganglia as well as MCP-1 in spinal dorsal horn. Whereas the antinociceptive effects of high dose JZL184 (40 mg/kg) underwent tolerance following 6 days of repeated dosing, repeated administration of a threshold dose (i.e., 4 mg/kg) completely reversed paclitaxel-induced allodynia. In addition, we found that the administration of MJN110 to control mice lacked intrinsic rewarding effects in the conditioned place preference (CPP) paradigm. However, it produced a CPP in paclitaxel-treated animals, suggesting a reduced paclitaxel-induced aversive state. Importantly, JZL184 did not alter the antiproliferative and apoptotic effects of paclitaxel in A549 and H460 non-small cell lung cancer cells. Taken together, these data indicate that MAGL inhibitors reverse paclitaxel-induced neuropathic pain without interfering with chemotherapeutic efficacy.
ESTHER : Curry_2018_J.Pharmacol.Exp.Ther_366_169
PubMedSearch : Curry_2018_J.Pharmacol.Exp.Ther_366_169
PubMedID: 29540562

Title : Monoacylglycerol Lipase Inhibition in Human and Rodent Systems Supports Clinical Evaluation of Endocannabinoid Modulators - Clapper_2018_J.Pharmacol.Exp.Ther_367_494
Author(s) : Clapper JR , Henry CL , Niphakis MJ , Knize AM , Coppola AR , Simon GM , Ngo N , Herbst RA , Herbst DM , Reed AW , Cisar JS , Weber OD , Viader A , Alexander JP , Cunningham ML , Jones TK , Fraser IP , Grice CA , Ezekowitz RAB , O'Neill GP , Blankman JL
Ref : Journal of Pharmacology & Experimental Therapeutics , 367 :494 , 2018
Abstract : Monoacylglycerol lipase (MGLL) is the primary degradative enzyme for the endocannabinoid 2-arachidonoylglycerol (2-AG). The first MGLL inhibitors have recently entered clinical development for the treatment of neurologic disorders. To support this clinical path, we report the pharmacological characterization of the highly potent and selective MGLL inhibitor ABD-1970 [1,1,1,3,3,3-hexafluoropropan-2-yl 4-(2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)-4-chlorobenzyl)piperazine-1-carboxylate]. We used ABD-1970 to confirm the role of MGLL in human systems and to define the relationship between MGLL target engagement, brain 2-AG concentrations, and efficacy. Because MGLL contributes to arachidonic acid metabolism in a subset of rodent tissues, we further used ABD-1970 to evaluate whether selective MGLL inhibition would affect prostanoid production in several human assays known to be sensitive to cyclooxygenase inhibitors. ABD-1970 robustly elevated brain 2-AG content and displayed antinociceptive and antipruritic activity in a battery of rodent models (ED(50) values of 1-2 mg/kg). The antinociceptive effects of ABD-1970 were potentiated when combined with analgesic standards of care and occurred without overt cannabimimetic effects. ABD-1970 also blocked 2-AG hydrolysis in human brain tissue and elevated 2-AG content in human blood without affecting stimulated prostanoid production. These findings support the clinical development of MGLL inhibitors as a differentiated mechanism to treat pain and other neurologic disorders.
ESTHER : Clapper_2018_J.Pharmacol.Exp.Ther_367_494
PubMedSearch : Clapper_2018_J.Pharmacol.Exp.Ther_367_494
PubMedID: 30305428

Title : Inhibition of the endocannabinoid-regulating enzyme monoacylglycerol lipase elicits a CB1 receptor-mediated discriminative stimulus in mice - Owens_2017_Neuropharmacol_125_80
Author(s) : Owens RA , Mustafa MA , Ignatowska-Jankowska BM , Damaj MI , Beardsley PM , Wiley JL , Niphakis MJ , Cravatt BF , Lichtman AH
Ref : Neuropharmacology , 125 :80 , 2017
Abstract : Substantial challenges exist for investigating the cannabinoid receptor type 1 (CB1)-mediated discriminative stimulus effects of the endocannabinoids, 2-arachidonoylglycerol (2-AG) and N-arachidonoylethanolamine (anandamide; AEA), compared with exogenous CB1 receptor agonists, such as Delta(9)-tetrahydrocannabinol (THC) and the synthetic cannabinoid CP55,940. Specifically, each endocannabinoid is rapidly degraded by the respective hydrolytic enzymes, monoacylglycerol lipase (MAGL) and fatty acid amide hydrolase (FAAH). Whereas MAGL inhibitors partially substitute for THC and fully substitute for CP55,940, FAAH inhibitors do not substitute for either drug. Interestingly, combined FAAH-MAGL inhibition results in full THC substitution, and the dual FAAH-MAGL inhibitor SA-57 serves as its own discriminative training stimulus. Because MAGL inhibitors fully substitute for SA-57, we tested whether the selective MAGL inhibitor MJN110 would serve as a training stimulus. Twelve of 13 C57BL/6J mice learned to discriminate MJN110 from vehicle, and the CB1 receptor antagonist rimonabant dose-dependently blocked its discriminative stimulus. CP55,940, SA-57, and another MAGL inhibitor JZL184, fully substituted for MJN110. In contrast, the FAAH inhibitor PF-3845 failed to substitute for the MJN110 discriminative stimulus, but produced a 1.6 (1.1-2.2; 95% confidence interval) leftward shift in the MJN110 dose-response curve. Inhibitors of other relevant enzymes (i.e., ABHD6, COX-2) and nicotine did not engender substitution. Diazepam partially substituted for MJN110, but rimonabant failed to block this partial effect. These findings suggest that MAGL normally throttles 2-AG stimulation of CB1 receptors to a magnitude insufficient to produce cannabimimetic subjective effects. Accordingly, inhibitors of this enzyme may release this endogenous brake producing effects akin to those produced by exogenously administered cannabinoids.
ESTHER : Owens_2017_Neuropharmacol_125_80
PubMedSearch : Owens_2017_Neuropharmacol_125_80
PubMedID: 28673548

Title : The endocannabinoid hydrolysis inhibitor SA-57: Intrinsic antinociceptive effects, augmented morphine-induced antinociception, and attenuated heroin seeking behavior in mice - Wilkerson_2017_Neuropharmacol_114_156
Author(s) : Wilkerson JL , Ghosh S , Mustafa M , Abdullah RA , Niphakis MJ , Cabrera R , Maldonado RL , Cravatt BF , Lichtman AH
Ref : Neuropharmacology , 114 :156 , 2017
Abstract : Although opioids are highly efficacious analgesics, their abuse potential and other untoward side effects diminish their therapeutic utility. The addition of non-opioid analgesics offers a promising strategy to reduce required antinociceptive opioid doses that concomitantly reduce opioid-related side effects. Inhibitors of the primary endocannabinoid catabolic enzymes fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) show opioid-sparing effects in preclinical models of pain. As simultaneous inhibition of these enzymes elicits enhanced antinociceptive effects compared with single enzyme inhibition, the present study tested whether the dual FAAH-MAGL inhibitor SA-57 [4-[2-(4-chlorophenyl)ethyl]-1-piperidinecarboxylic acid 2-(methylamino)-2-oxoethyl ester] produces morphine-sparing antinociceptive effects, without major side effects associated with either drug class. SA-57 dose-dependently reversed mechanical allodynia in the constriction injury (CCI) of the sciatic nerve model of neuropathic pain and carrageenan inflammatory pain model. As previously reported, SA-57 was considerably more potent in elevating anandamide (AEA) than 2-arachidonyl glycerol (2-AG) in brain. Its anti-allodynic effects required cannabinoid (CB)1 and CB2 receptors; however, only CB2 receptors were necessary for the anti-edematous effects in the carrageenan assay. Although high doses of SA-57 alone were required to produce antinociception, low doses of this compound, which elevated AEA and did not affect 2-AG brain levels, augmented the antinociceptive effects of morphine, but lacked cannabimimetic side effects. Because of the high abuse liability of opioids and implication of the endocannabinoid system in the reinforcing effects of opioids, the final experiment tested whether SA-57 would alter heroin seeking behavior. Strikingly, SA-57 reduced heroin-reinforced nose poke behavior and the progressive ratio break point for heroin. In conclusion, the results of the present study suggest that inhibition of endocannabinoid degradative enzymes represents a promising therapeutic approach to decrease effective doses of opioids needed for clinical pain control, and may also possess therapeutic potential to reduce opioid abuse.
ESTHER : Wilkerson_2017_Neuropharmacol_114_156
PubMedSearch : Wilkerson_2017_Neuropharmacol_114_156
PubMedID: 27890602

Title : Discriminative Stimulus Properties of the Endocannabinoid Catabolic Enzyme Inhibitor SA-57 in Mice - Owens_2016_J.Pharmacol.Exp.Ther_358_306
Author(s) : Owens RA , Ignatowska-Jankowska B , Mustafa M , Beardsley PM , Wiley JL , Jali A , Selley DE , Niphakis MJ , Cravatt BF , Lichtman AH
Ref : Journal of Pharmacology & Experimental Therapeutics , 358 :306 , 2016
Abstract : Whereas the inhibition of fatty acid amide hydrolase (FAAH) or monoacylglycerol lipase (MAGL), the respective major hydrolytic enzymes of N-arachidonoyl ethanolamine (AEA) and 2-arachidonoylglycerol (2-AG), elicits no or partial substitution for Delta(9)-tetrahydrocannabinol (THC) in drug-discrimination procedures, combined inhibition of both enzymes fully substitutes for THC, as well as produces a constellation of cannabimimetic effects. The present study tested whether C57BL/6J mice would learn to discriminate the dual FAAH-MAGL inhibitor SA-57 (4-[2-(4-chlorophenyl)ethyl]-1-piperidinecarboxylic acid 2-(methylamino)-2-oxoethyl ester) from vehicle in the drug-discrimination paradigm. In initial experiments, 10 mg/kg SA-57 fully substituted for CP55,940 ((-)-cis-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-trans-4-(3-hydroxypropyl)cycl ohexanol), a high-efficacy CB1 receptor agonist in C57BL/6J mice and for AEA in FAAH (-/-) mice. Most (i.e., 23 of 24) subjects achieved criteria for discriminating SA-57 (10 mg/kg) from vehicle within 40 sessions, with full generalization occurring 1 to 2 hours postinjection. CP55,940, the dual FAAH-MAGL inhibitor JZL195 (4-nitrophenyl 4-(3-phenoxybenzyl)piperazine-1-carboxylate), and the MAGL inhibitors MJN110 (2,5-dioxopyrrolidin-1-yl 4-(bis(4-chlorophenyl)methyl)piperazine-1-carboxylate) and JZL184 (4-[Bis(1,3-benzodioxol-5-yl)hydroxymethyl]-1-piperidinecarboxylic acid 4-nitrophenyl ester) fully substituted for SA-57. Although the FAAH inhibitors PF-3845 ((N-3-pyridinyl-4-[[3-[[5-(trifluoromethyl)-2-pyridinyl]oxy]phenyl]methyl]-1-pipe ridinecarboxamide) and URB597 (cyclohexylcarbamic acid 3'-(aminocarbonyl)-[1,1'-biphenyl]-3-yl ester) did not substitute for SA-57, PF-3845 produced a 2-fold leftward shift in the MJN110 substitution dose-response curve. In addition, the CB1 receptor antagonist rimonabant blocked the generalization of SA-57, as well as substitution of CP55,940, JZL195, MJN110, and JZL184. These findings suggest that MAGL inhibition plays a major role in the CB1 receptor-mediated SA-57 training dose, which is further augmented by FAAH inhibition.
ESTHER : Owens_2016_J.Pharmacol.Exp.Ther_358_306
PubMedSearch : Owens_2016_J.Pharmacol.Exp.Ther_358_306
PubMedID: 27307500

Title : Elevation of 2-AG by monoacylglycerol lipase inhibition in the visceral insular cortex interferes with anticipatory nausea in a rat model - Limebeer_2016_Behav.Neurosci_130_261
Author(s) : Limebeer CL , Rock EM , Puvanenthirarajah N , Niphakis MJ , Cravatt BF , Parker LA
Ref : Behavioral Neuroscience , 130 :261 , 2016
Abstract : Anticipatory nausea (AN) is a conditioned nausea reaction experienced by chemotherapy patients upon returning to the clinic. Currently, there are no specific treatments for this phenomenon, with the classic antiemetic treatments (e.g., ondansetron) providing no relief. The rat model of AN, contextually elicited conditioned gaping reactions in rats, provides a tool for assessing potential treatments for this difficult to treat disorder. Systemically administered drugs which elevate the endocannabinoids, anandamide (AEA) and 2-arachidonoyl glycerol (2-AG), by interfering with their respective degrading enzymes, fatty acid amide hydrolase (FAAH) and monoacyl glycerol lipase (MAGL) interfere with AN in the rat model. We have shown that MAGL inhibition within the visceral insular cortex (VIC) interferes with acute nausea in the gaping model (Sticht et al., 2015). Here we report that bilateral infusion of the MAGL inhibitor, MJN110 (but neither the FAAH inhibitor, PF3845, nor ondansetron) into the VIC suppressed contextually elicited conditioned gaping, and this effect was reversed by coadministration of the CB1 antagonist, AM251. These findings suggest that 2-AG within the VIC plays a critical role in the regulation of both acute nausea and AN. Because there are currently no specific therapeutics for chemotherapy patients that develop anticipatory nausea, MAGL inhibition by MJN110 may be a candidate treatment. (PsycINFO Database Record
ESTHER : Limebeer_2016_Behav.Neurosci_130_261
PubMedSearch : Limebeer_2016_Behav.Neurosci_130_261
PubMedID: 26974857

Title : The Selective Monoacylglycerol Lipase Inhibitor MJN110 Produces Opioid-Sparing Effects in a Mouse Neuropathic Pain Model - Wilkerson_2016_J.Pharmacol.Exp.Ther_357_145
Author(s) : Wilkerson JL , Niphakis MJ , Grim TW , Mustafa MA , Abdullah RA , Poklis JL , Dewey WL , Akbarali H , Banks ML , Wise LE , Cravatt BF , Lichtman AH
Ref : Journal of Pharmacology & Experimental Therapeutics , 357 :145 , 2016
Abstract : Serious clinical liabilities associated with the prescription of opiates for pain control include constipation, respiratory depression, pruritus, tolerance, abuse, and addiction. A recognized strategy to circumvent these side effects is to combine opioids with other antinociceptive agents. The combination of opiates with the primary active constituent of cannabis (Delta(9)-tetrahydrocannabinol) produces enhanced antinociceptive actions, suggesting that cannabinoid receptor agonists can be opioid sparing. Here, we tested whether elevating the endogenous cannabinoid 2-arachidonoylglycerol through the inhibition of its primary hydrolytic enzyme monoacylglycerol lipase (MAGL), will produce opioid-sparing effects in the mouse chronic constriction injury (CCI) of the sciatic nerve model of neuropathic pain. The dose-response relationships of i.p. administration of morphine and the selective MAGL inhibitor 2,5-dioxopyrrolidin-1-yl 4-(bis(4-chlorophenyl)methyl)piperazine-1-carboxylate (MJN110) were tested alone and in combination at equieffective doses for reversal of CCI-induced mechanical allodynia and thermal hyperalgesia. The respective ED50 doses (95% confidence interval) of morphine and MJN110 were 2.4 (1.9-3.0) mg/kg and 0.43 (0.23-0.79) mg/kg. Isobolographic analysis of these drugs in combination revealed synergistic antiallodynic effects. Acute antinociceptive effects of the combination of morphine and MJN110 required mu-opioid, CB1, and CB2 receptors. This combination did not reduce gastric motility or produce subjective cannabimimetic effects in the drug discrimination assay. Importantly, combinations of MJN110 and morphine given repeatedly (i.e., twice a day for 6 days) continued to produce antiallodynic effects with no evidence of tolerance. Taken together, these findings suggest that MAGL inhibition produces opiate-sparing events with diminished tolerance, constipation, and cannabimimetic side effects.
ESTHER : Wilkerson_2016_J.Pharmacol.Exp.Ther_357_145
PubMedSearch : Wilkerson_2016_J.Pharmacol.Exp.Ther_357_145
PubMedID: 26791602

Title : Robust anti-nociceptive effects of monoacylglycerol lipase inhibition in a model of osteoarthritis pain - Burston_2016_Br.J.Pharmacol_173_3134
Author(s) : Burston JJ , Mapp PI , Sarmad S , Barrett DA , Niphakis MJ , Cravatt BF , Walsh DA , Chapman V
Ref : British Journal of Pharmacology , 173 :3134 , 2016
Abstract : BACKGROUND AND PURPOSE: Chronic pain is often a symptom of knee osteoarthritis (OA) for which current analgesics are either inadequate or are associated with serious side effects. The endocannabinoid system may offer alternative targets for pain relief. We evaluated the effects of a potent and selective monoacylglycerol (MAG) lipase inhibitor (MJN110) on OA pain behaviour, spinal mechanisms of action and joint histopathology in the rat. EXPERIMENTAL APPROACH: Intra-articular injection of monosodium iodoacetate (MIA) models OA pain and mimics clinical joint pathology. Effects of MJN110 on MIA-induced weight-bearing asymmetry and lowered paw withdrawal thresholds (PWTs), changes in spinal gene expression and brain levels of relevant lipids were determined. KEY
RESULTS: Acute MJN110 (5 ) significantly reversed MIA-induced weight-bearing asymmetry (MIA/vehicle: 68 +/- 6 g; MIA/MJN110: 35 +/- 4 g) and lowered ipsilateral PWTs (MIA/vehicle: 7 +/- 0.8 g; MIA/MJN110: 11 +/- 0.6 g), via both CB1 and CB2 receptors. Repeated treatment with MJN110 (5 ) resulted in anti-nociceptive tolerance. A lower dose of MJN110 (1 ) acutely inhibited pain behaviour, which was maintained for 1 week of repeated administration but had no effect on joint histology. MJN110 significantly inhibited expression of membrane-associated PGE synthase-1 in the ipsilateral dorsal horn of the spinal cord of MIA rats, compared with vehicle-treated MIA rats. Both doses of MJN110 significantly elevated brain levels of the endocannabinoid 2-arachidonoylglycerol. CONCLUSIONS AND IMPLICATIONS: Our data support further assessment of the therapeutic potential of MAG lipase inhibitors for the treatment of OA pain.
ESTHER : Burston_2016_Br.J.Pharmacol_173_3134
PubMedSearch : Burston_2016_Br.J.Pharmacol_173_3134
PubMedID: 27501482

Title : Double Dissociation of Monoacylglycerol Lipase Inhibition and CB1 Antagonism in the Central Amygdala, Basolateral Amygdala, and the Interoceptive Insular Cortex on the Affective Properties of Acute Naloxone-Precipitated Morphine Withdrawal in Rats - Wills_2016_Neuropsychopharmacology_41_1865
Author(s) : Wills KL , Petrie GN , Millett G , Limebeer CL , Rock EM , Niphakis MJ , Cravatt BF , Parker LA
Ref : Neuropsychopharmacology , 41 :1865 , 2016
Abstract : Both CB1 receptor antagonism and agonism, in particular by 2-arachidonyl glycerol (2-AG), have been shown to reduce somatic symptoms of morphine withdrawal (MWD). Here we evaluated the effects of both systemic pretreatment with the monoacylglycerol lipase (MAGL) inhibitor MJN110 (which selectively elevates 2-AG) and central administration of both MJN110 and the CB1 antagonist (AM251) on the affective properties of MWD. Acute MWD induced place aversion occurs when naloxone is administered 24 h following a single exposure to a high dose of morphine. Systemic pretreatment with the MAGL inhibitor, MJN110, prevented the aversive effects of acute MWD by a CB1 receptor-dependent mechanism. Furthermore, in a double dissociation, AM251 infusions into the central amygdala, but MJN110 infusions into the basolateral amygdala, interfered with the naloxone-precipitated MWD induced place aversion. As well, MJN110, but not AM251, infusions into the interoceptive insular cortex (a region known to be activated in acute MWD) also prevented the establishment of the place aversion by a CB1 mechanism of action. These findings reveal the respective sites of action of systemically administered MJN110 and AM251 in regulating the aversive effects of MWD.
ESTHER : Wills_2016_Neuropsychopharmacology_41_1865
PubMedSearch : Wills_2016_Neuropsychopharmacology_41_1865
PubMedID: 26647976

Title : Endocannabinoid regulation of nausea is mediated by 2-arachidonoylglycerol (2-AG) in the rat visceral insular cortex - Sticht_2016_Neuropharmacol_102_92
Author(s) : Sticht MA , Limebeer CL , Rafla BR , Abdullah RA , Poklis JL , Ho W , Niphakis MJ , Cravatt BF , Sharkey KA , Lichtman AH , Parker LA
Ref : Neuropharmacology , 102 :92 , 2016
Abstract : Cannabinoid (CB) agonists suppress nausea in humans and animal models; yet, their underlying neural substrates remain largely unknown. Evidence suggests that the visceral insular cortex (VIC) plays a critical role in nausea. Given the expression of CB1 receptors and the presence of endocannabinoids in this brain region, we hypothesized that the VIC endocannabinoid system regulates nausea. In the present study, we assessed whether inhibiting the primary endocannabinoid hydrolytic enzymes in the VIC reduces acute lithium chloride (LiCl)-induced conditioned gaping, a rat model of nausea. We also quantified endocannabinoid levels during an episode of nausea, and assessed VIC neuronal activation using the marker, c-Fos. Local inhibition of monoacylglycerol lipase (MAGL), the main hydrolytic enzyme of 2-arachidonylglycerol (2-AG), reduced acute nausea through a CB1 receptor mechanism, whereas inhibition of fatty acid amide hydrolase (FAAH), the primary catabolic enzyme of anandamide (AEA), was without effect. Levels of 2-AG were also selectively elevated in the VIC during an episode of nausea. Inhibition of MAGL robustly increased 2-AG in the VIC, while FAAH inhibition had no effect on AEA. Finally, we demonstrated that inhibition of MAGL reduced VIC Fos immunoreactivity in response to LiCl treatment. Taken together, these findings provide compelling evidence that acute nausea selectively increases 2-AG in the VIC, and suggests that 2-AG signaling within the VIC regulates nausea by reducing neuronal activity in this forebrain region.
ESTHER : Sticht_2016_Neuropharmacol_102_92
PubMedSearch : Sticht_2016_Neuropharmacol_102_92
PubMedID: 26541329

Title : Diacylglycerol lipase disinhibits VTA dopamine neurons during chronic nicotine exposure - Buczynski_2016_Proc.Natl.Acad.Sci.U.S.A_113_1086
Author(s) : Buczynski MW , Herman MA , Hsu KL , Natividad LA , Irimia C , Polis IY , Pugh H , Chang JW , Niphakis MJ , Cravatt BF , Roberto M , Parsons LH
Ref : Proc Natl Acad Sci U S A , 113 :1086 , 2016
Abstract : Chronic nicotine exposure (CNE) alters synaptic transmission in the ventral tegmental area (VTA) in a manner that enhances dopaminergic signaling and promotes nicotine use. The present experiments identify a correlation between enhanced production of the endogenous cannabinoid 2-arachidonoylglycerol (2-AG) and diminished release of the inhibitory neurotransmitter GABA in the VTA following CNE. To study the functional role of on-demand 2-AG signaling in GABAergic synapses, we used 1,2,3-triazole urea compounds to selectively inhibit 2-AG biosynthesis by diacylglycerol lipase (DAGL). The potency and selectivity of these inhibitors were established in rats in vitro (rat brain proteome), ex vivo (brain slices), and in vivo (intracerebroventricular administration) using activity-based protein profiling and targeted metabolomics analyses. Inhibition of DAGL (2-AG biosynthesis) rescues nicotine-induced VTA GABA signaling following CNE. Conversely, enhancement of 2-AG signaling in naive rats by inhibiting 2-AG degradation recapitulates the loss of nicotine-induced GABA signaling evident following CNE. DAGL inhibition reduces nicotine self-administration without disrupting operant responding for a nondrug reinforcer or motor activity. Collectively, these findings provide a detailed characterization of selective inhibitors of rat brain DAGL and demonstrate that excessive 2-AG signaling contributes to a loss of inhibitory GABAergic constraint of VTA excitability following CNE.
ESTHER : Buczynski_2016_Proc.Natl.Acad.Sci.U.S.A_113_1086
PubMedSearch : Buczynski_2016_Proc.Natl.Acad.Sci.U.S.A_113_1086
PubMedID: 26755579

Title : Delta9-tetrahydrocannabinol and endocannabinoid degradative enzyme inhibitors attenuate intracranial self-stimulation in mice - Wiebelhaus_2015_J.Pharmacol.Exp.Ther_352_195
Author(s) : Wiebelhaus JM , Grim TW , Owens RA , Lazenka MF , Sim-Selley LJ , Abdullah RA , Niphakis MJ , Vann RE , Cravatt BF , Wiley JL , Negus SS , Lichtman AH
Ref : Journal of Pharmacology & Experimental Therapeutics , 352 :195 , 2015
Abstract : A growing body of evidence implicates endogenous cannabinoids as modulators of the mesolimbic dopamine system and motivated behavior. Paradoxically, the reinforcing effects of Delta(9)-tetrahydrocannabinol (THC), the primary psychoactive constituent of cannabis, have been difficult to detect in preclinical rodent models. In this study, we investigated the impact of THC and inhibitors of the endocannabinoid hydrolytic enzymes fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) on operant responding for electrical stimulation of the medial forebrain bundle [intracranial self-stimulation (ICSS)], which is known to activate the mesolimbic dopamine system. These drugs were also tested in assays of operant responding for food reinforcement and spontaneous locomotor activity. THC and the MAGL inhibitor JZL184 (4-[bis(1,3-benzodioxol-5-yl)hydroxymethyl]-1-piperidinecarboxylic acid 4-nitrophenyl ester) attenuated operant responding for ICSS and food, and also reduced spontaneous locomotor activity. In contrast, the FAAH inhibitor PF-3845 (N-3-pyridinyl-4-[[3-[[5-(trifluoromethyl)-2-pyridinyl]oxy]phenyl]methyl]-1-piper idinecarboxamide) was largely without effect in these assays. Consistent with previous studies showing that combined inhibition of FAAH and MAGL produces a substantially greater cannabimimetic profile than single enzyme inhibition, the dual FAAH-MAGL inhibitor SA-57 (4-[2-(4-chlorophenyl)ethyl]-1-piperidinecarboxylic acid 2-(methylamino)-2-oxoethyl ester) produced a similar magnitude of ICSS depression as that produced by THC. ICSS attenuation by JZL184 was associated with increased brain levels of 2-arachidonoylglycerol (2-AG), whereas peak effects of SA-57 were associated with increased levels of both N-arachidonoylethanolamine (anandamide) and 2-AG. The cannabinoid receptor type 1 receptor antagonist rimonabant, but not the cannabinoid receptor type 2 receptor antagonist SR144528, blocked the attenuating effects of THC, JZL184, and SA-57 on ICSS. Thus, THC, MAGL inhibition, and dual FAAH-MAGL inhibition not only reduce ICSS, but also decrease other reinforced and nonreinforced behaviors.
ESTHER : Wiebelhaus_2015_J.Pharmacol.Exp.Ther_352_195
PubMedSearch : Wiebelhaus_2015_J.Pharmacol.Exp.Ther_352_195
PubMedID: 25398241

Title : Selective N-Hydroxyhydantoin Carbamate Inhibitors of Mammalian Serine Hydrolases - Cognetta_2015_Chem.Biol_22_928
Author(s) : Cognetta AB, 3rd , Niphakis MJ , Lee HC , Martini ML , Hulce JJ , Cravatt BF
Ref : Chemical Biology , 22 :928 , 2015
Abstract : Serine hydrolase inhibitors, which facilitate enzyme function assignment and are used to treat a range of human disorders, often act by an irreversible mechanism that involves covalent modification of the serine hydrolase catalytic nucleophile. The portion of mammalian serine hydrolases for which selective inhibitors have been developed, however, remains small. Here, we show that N-hydroxyhydantoin (NHH) carbamates are a versatile class of irreversible serine hydrolase inhibitors that can be modified on both the staying (carbamylating) and leaving (NHH) groups to optimize potency and selectivity. Synthesis of a small library of NHH carbamates and screening by competitive activity-based protein profiling furnished selective, in vivo-active inhibitors and tailored activity-based probes for multiple mammalian serine hydrolases, including palmitoyl protein thioesterase 1, mutations of which cause the human disease infantile neuronal ceroid lipofuscinosis.
ESTHER : Cognetta_2015_Chem.Biol_22_928
PubMedSearch : Cognetta_2015_Chem.Biol_22_928
PubMedID: 26120000

Title : Effect of selective inhibition of monoacylglycerol lipase (MAGL) on acute nausea, anticipatory nausea, and vomiting in rats and Suncus murinus - Parker_2015_Psychopharmacology.(Berl)_232_583
Author(s) : Parker LA , Niphakis MJ , Downey R , Limebeer CL , Rock EM , Sticht MA , Morris H , Abdullah RA , Lichtman AH , Cravatt BF
Ref : Psychopharmacology (Berl) , 232 :583 , 2015
Abstract : RATIONALE: To determine the role of the endocannabinoid, 2-arachodonyl glycerol (2-AG), in the regulation of nausea and vomiting. OBJECTIVE: We evaluated the effectiveness of the potent selective monoacylglycerol lipase (MAGL) inhibitor, MJN110, which selectively elevates the endocannabinoid 2-AG, to suppress acute nausea and vomiting, as well as anticipatory nausea in rat and shrew models.
METHODS: The rat gaping models were used to evaluate the potential of MJN110 (5, 10, and 20 mg/kg, intraperitoneally [IP]) to suppress acute nausea produced by LiCl and of MJN110 (10 and 20 mg/kg, IP) to suppress anticipatory nausea elicited by a LiCl-paired context. The potential as well of MJN110 (10 and 20 mg/kg, IP) to suppress vomiting and contextually elicited gaping in the Suncus murinus was evaluated.
RESULTS: MJN110 suppressed acute nausea in rats, LiCl-induced vomiting in shrews and contextually-elicited anticipatory nausea in both rats (accompanied by elevation of 2-AG in the visceral insular cortex) and shrews. These effects were reversed by the CB1 antagonist/inverse agonist, SR141716. The MAGL inhibitor did not modify locomotion at any dose. An activity-based protein profiling analysis of samples of tissue collected from the visceral insular cortex in rats and whole brain tissues in shrews revealed that MJN110 selectively inhibited MAGL and the alternative 2-AG hydrolase, ABHD6.
CONCLUSIONS: MAGL inhibition by MJN110 which selectively elevates endogenous 2-AG has therapeutic potential in the treatment of acute nausea and vomiting as well as anticipatory nausea, a distressful symptom that is resistant to currently available treatments.
ESTHER : Parker_2015_Psychopharmacology.(Berl)_232_583
PubMedSearch : Parker_2015_Psychopharmacology.(Berl)_232_583
PubMedID: 25085768
Gene_locus related to this paper: human-MGLL

Title : In vivo characterization of the highly selective monoacylglycerol lipase inhibitor KML29: antinociceptive activity without cannabimimetic side effects - Ignatowska-Jankowska_2014_Br.J.Pharmacol_171_1392
Author(s) : Ignatowska-Jankowska BM , Ghosh S , Crowe MS , Kinsey SG , Niphakis MJ , Abdullah RA , Tao Q , ST ON , Walentiny DM , Wiley JL , Cravatt BF , Lichtman AH
Ref : British Journal of Pharmacology , 171 :1392 , 2014
Abstract : BACKGROUND AND PURPOSE: Since monoacylglycerol lipase (MAGL) has been firmly established as the predominant catabolic enzyme of the endocannabinoid 2-arachidonoylglycerol (2-AG), a great need has emerged for the development of highly selective MAGL inhibitors. Here, we tested the in vivo effects of one such compound, KML29 (1,1,1,3,3,3-hexafluoropropan-2-yl 4-(bis(benzo[d][1,3]dioxol-5-yl)(hydroxy)methyl)piperidine-1-carboxylate). EXPERIMENTAL APPROACH: In the present study, we tested KML29 in murine inflammatory (i.e. carrageenan) and sciatic nerve injury pain models, as well as the diclofenac-induced gastric haemorrhage model. KML29 was also evaluated for cannabimimetic effects, including measurements of locomotor activity, body temperature, catalepsy, and cannabinoid interoceptive effects in the drug discrimination paradigm. KEY
RESULTS: KML29 attenuated carrageenan-induced paw oedema and completely reversed carrageenan-induced mechanical allodynia. These effects underwent tolerance after repeated administration of high-dose KML29, which were accompanied by cannabinoid receptor 1 (CB1 ) receptor desensitization. Acute or repeated KML29 administration increased 2-AG levels and concomitantly reduced arachidonic acid levels, but without elevating anandamide (AEA) levels in the whole brain. Furthermore, KML29 partially reversed allodynia in the sciatic nerve injury model and completely prevented diclofenac-induced gastric haemorrhages. CB1 and CB2 receptors played differential roles in these pharmacological effects of KML29. In contrast, KML29 did not elicit cannabimimetic effects, including catalepsy, hypothermia and hypomotility. Although KML29 did not substitute for Delta(9) -tetrahydrocannabinol (THC) in C57BL/6J mice, it fully and dose-dependantly substituted for AEA in fatty acid amide hydrolase (FAAH) (-/-) mice, consistent with previous work showing that dual FAAH and MAGL inhibition produces THC-like subjective effects. CONCLUSIONS AND IMPLICATIONS: These results indicate that KML29, a highly selective MAGL inhibitor, reduces inflammatory and neuropathic nociceptive behaviour without occurrence of cannabimimetic side effects. LINKED ARTICLES: This article is part of a themed section on Cannabinoids 2013. To view the other articles in this section visit
ESTHER : Ignatowska-Jankowska_2014_Br.J.Pharmacol_171_1392
PubMedSearch : Ignatowska-Jankowska_2014_Br.J.Pharmacol_171_1392
PubMedID: 23848221
Gene_locus related to this paper: human-MGLL

Title : Endocannabinoids modulate cortical development by configuring Slit2\/Robo1 signalling - Alpar_2014_Nat.Commun_5_4421
Author(s) : Alpar A , Tortoriello G , Calvigioni D , Niphakis MJ , Milenkovic I , Bakker J , Cameron GA , Hanics J , Morris CV , Fuzik J , Kovacs GG , Cravatt BF , Parnavelas JG , Andrews WD , Hurd YL , Keimpema E , Harkany T
Ref : Nat Commun , 5 :4421 , 2014
Abstract : Local environmental cues are indispensable for axonal growth and guidance during brain circuit formation. Here, we combine genetic and pharmacological tools, as well as systems neuroanatomy in human fetuses and mouse models, to study the role of endocannabinoid and Slit/Robo signalling in axonal growth. We show that excess 2-arachidonoylglycerol, an endocannabinoid affecting directional axonal growth, triggers corpus callosum enlargement due to the errant CB1 cannabinoid receptor-containing corticofugal axon spreading. This phenotype mechanistically relies on the premature differentiation and end-feet proliferation of CB2R-expressing oligodendrocytes. We further show the dependence of both axonal Robo1 positioning and oligodendroglial Slit2 production on cell-type-specific cannabinoid receptor activation. Accordingly, Robo1 and/or Slit2 manipulation limits endocannabinoid modulation of axon guidance. We conclude that endocannabinoids can configure focal Slit2/Robo1 signalling to modulate directional axonal growth, which may provide a basis for understanding impaired brain wiring associated with metabolic deficits and prenatal drug exposure.
ESTHER : Alpar_2014_Nat.Commun_5_4421
PubMedSearch : Alpar_2014_Nat.Commun_5_4421
PubMedID: 25030704

Title : Remodeling natural products: chemistry and serine hydrolase activity of a rocaglate-derived beta-lactone - Lajkiewicz_2014_J.Am.Chem.Soc_136_2659
Author(s) : Lajkiewicz NJ , Cognetta AB, 3rd , Niphakis MJ , Cravatt BF , Porco JA, Jr.
Ref : Journal of the American Chemical Society , 136 :2659 , 2014
Abstract : Flavaglines are a class of natural products with potent insecticidal and anticancer activities. beta-Lactones are a privileged structural motif found in both therapeutic agents and chemical probes. Herein, we report the synthesis, unexpected light-driven di-epimerization, and activity-based protein profiling of a novel rocaglate-derived beta-lactone. In addition to in vitro inhibition of the serine hydrolases ABHD10 and ACOT1/2, the most potent beta-lactone enantiomer was also found to inhibit these enzymes, as well as the serine peptidases CTSA and SCPEP1, in PC3 cells.
ESTHER : Lajkiewicz_2014_J.Am.Chem.Soc_136_2659
PubMedSearch : Lajkiewicz_2014_J.Am.Chem.Soc_136_2659
PubMedID: 24447064
Gene_locus related to this paper: human-ABHD10 , human-ACOT1 , human-ACOT2 , human-CTSA , human-SCPEP1 , mouse-abhda

Title : Mapping the protein interaction landscape for fully functionalized small-molecule probes in human cells - Kambe_2014_J.Am.Chem.Soc_136_10777
Author(s) : Kambe T , Correia BE , Niphakis MJ , Cravatt BF
Ref : Journal of the American Chemical Society , 136 :10777 , 2014
Abstract : Phenotypic screening provides a means to discover small molecules that perturb cell biological processes. Discerning the proteins and biochemical pathways targeted by screening hits, however, remains technically challenging. We recently described the use of small molecules bearing photoreactive groups and latent affinity handles as fully functionalized probes for integrated phenotypic screening and target identification. The general utility of such probes, or, for that matter, any small-molecule screening library, depends on the scope of their protein interactions in cells, a parameter that remains largely unexplored. Here, we describe the synthesis of an ~60-member fully functionalized probe library, prepared from Ugi-azide condensation reactions to impart structural diversity and introduce diazirine and alkyne functionalities for target capture and enrichment, respectively. In-depth mass spectrometry-based analysis revealed a diverse array of probe targets in human cells, including enzymes, channels, adaptor and scaffolding proteins, and proteins of uncharacterized function. For many of these proteins, ligands have not yet been described. Most of the probe-protein interactions showed well-defined structure-activity relationships across the probe library and were blocked by small-molecule competitors in cells. These findings indicate that fully functionalized small molecules canvas diverse segments of the human proteome and hold promise as pharmacological probes of cell biology.
ESTHER : Kambe_2014_J.Am.Chem.Soc_136_10777
PubMedSearch : Kambe_2014_J.Am.Chem.Soc_136_10777
PubMedID: 25045785

Title : Enzyme inhibitor discovery by activity-based protein profiling - Niphakis_2014_Annu.Rev.Biochem_83_341
Author(s) : Niphakis MJ , Cravatt BF
Ref : Annual Review of Biochemistry , 83 :341 , 2014
Abstract : Eukaryotic and prokaryotic organisms possess huge numbers of uncharacterized enzymes. Selective inhibitors offer powerful probes for assigning functions to enzymes in native biological systems. Here, we discuss how the chemical proteomic platform activity-based protein profiling (ABPP) can be implemented to discover selective and in vivo-active inhibitors for enzymes. We further describe how these inhibitors have been used to delineate the biochemical and cellular functions of enzymes, leading to the discovery of metabolic and signaling pathways that make important contributions to human physiology and disease. These studies demonstrate the value of selective chemical probes as drivers of biological inquiry.
ESTHER : Niphakis_2014_Annu.Rev.Biochem_83_341
PubMedSearch : Niphakis_2014_Annu.Rev.Biochem_83_341
PubMedID: 24905785

Title : Proteome-wide mapping of cholesterol-interacting proteins in mammalian cells - Hulce_2013_Nat.Methods_10_259
Author(s) : Hulce JJ , Cognetta AB, 3rd , Niphakis MJ , Tully SE , Cravatt BF
Ref : Nat Methods , 10 :259 , 2013
Abstract : Cholesterol is an essential structural component of cellular membranes and serves as a precursor for several classes of signaling molecules. Cholesterol exerts its effects and is, itself, regulated in large part by engagement in specific interactions with proteins. The full complement of sterol-binding proteins that exist in mammalian cells, however, remains unknown. Here we describe a chemoproteomic strategy that uses clickable, photoreactive sterol probes in combination with quantitative mass spectrometry to globally map cholesterol-protein interactions directly in living cells. We identified over 250 cholesterol-binding proteins, including receptors, channels and enzymes involved in many established and previously unreported interactions. Prominent among the newly identified interacting proteins were enzymes that regulate sugars, glycerolipids and cholesterol itself as well as proteins involved in vesicular transport and protein glycosylation and degradation, pointing to key nodes in biochemical pathways that may couple sterol concentrations to the control of other metabolites and protein localization and modification.
ESTHER : Hulce_2013_Nat.Methods_10_259
PubMedSearch : Hulce_2013_Nat.Methods_10_259
PubMedID: 23396283

Title : Selective inhibitors and tailored activity probes for lipoprotein-associated phospholipase A(2) - Nagano_2013_Bioorg.Med.Chem.Lett_23_839
Author(s) : Nagano JM , Hsu KL , Whitby LR , Niphakis MJ , Speers AE , Brown SJ , Spicer T , Fernandez-Vega V , Ferguson J , Hodder P , Srinivasan P , Gonzalez TD , Rosen H , Bahnson BJ , Cravatt BF
Ref : Bioorganic & Medicinal Chemistry Lett , 23 :839 , 2013
Abstract : Lipoprotein-associated phospholipase A(2) (Lp-PLA(2) or PLA(2)G7) binds to low-density lipoprotein (LDL) particles, where it is thought to hydrolyze oxidatively truncated phospholipids. Lp-PLA(2) has also been implicated as a pro-tumorigenic enzyme in human prostate cancer. Several inhibitors of Lp-PLA(2) have been described, including darapladib, which is currently in phase 3 clinical development for the treatment of atherosclerosis. The selectivity that darapladib and other Lp-PLA(2) inhibitors display across the larger serine hydrolase family has not, however, been reported. Here, we describe the use of both general and tailored activity-based probes for profiling Lp-PLA(2) and inhibitors of this enzyme in native biological systems. We show that both darapladib and a novel class of structurally distinct carbamate inhibitors inactivate Lp-PLA(2) in mouse tissues and human cell lines with high selectivity. Our findings thus identify both inhibitors and chemoproteomic probes that are suitable for investigating Lp-PLA(2) function in biological systems.
ESTHER : Nagano_2013_Bioorg.Med.Chem.Lett_23_839
PubMedSearch : Nagano_2013_Bioorg.Med.Chem.Lett_23_839
PubMedID: 23260346

Title : Proteome-wide reactivity profiling identifies diverse carbamate chemotypes tuned for serine hydrolase inhibition - Chang_2013_ACS.Chem.Biol_8_1590
Author(s) : Chang JW , Cognetta AB, 3rd , Niphakis MJ , Cravatt BF
Ref : ACS Chemical Biology , 8 :1590 , 2013
Abstract : Serine hydrolases are one of the largest and most diverse enzyme classes in Nature. Inhibitors of serine hydrolases are used to treat many diseases, including obesity, diabetes, cognitive dementia, and bacterial and viral infections. Nonetheless, the majority of the 200+ serine hydrolases in mammals still lack selective inhibitors for their functional characterization. We and others have shown that activated carbamates, through covalent reaction with the conserved serine nucleophile of serine hydrolases, can serve as useful inhibitors for members of this enzyme family. The extent to which carbamates, however, cross-react with other protein classes remains mostly unexplored. Here, we address this problem by investigating the proteome-wide reactivity of a diverse set of activated carbamates in vitro and in vivo, using a combination of competitive and click chemistry (CC)-activity-based protein profiling (ABPP). We identify multiple classes of carbamates, including O-aryl, O-hexafluoroisopropyl (HFIP), and O-N-hydroxysuccinimidyl (NHS) carbamates that react selectively with serine hydrolases across entire mouse tissue proteomes in vivo. We exploit the proteome-wide specificity of HFIP carbamates to create in situ imaging probes for the endocannabinoid hydrolases monoacylglycerol lipase (MAGL) and alpha-beta hydrolase-6 (ABHD6). These findings, taken together, designate the carbamate as a privileged reactive group for serine hydrolases that can accommodate diverse structural modifications to produce inhibitors that display exceptional potency and selectivity across the mammalian proteome.
ESTHER : Chang_2013_ACS.Chem.Biol_8_1590
PubMedSearch : Chang_2013_ACS.Chem.Biol_8_1590
PubMedID: 23701408

Title : Evaluation of NHS carbamates as a potent and selective class of endocannabinoid hydrolase inhibitors - Niphakis_2013_ACS.Chem.Neurosci_4_1322
Author(s) : Niphakis MJ , Cognetta AB, 3rd , Chang JW , Buczynski MW , Parsons LH , Byrne F , Burston JJ , Chapman V , Cravatt BF
Ref : ACS Chem Neurosci , 4 :1322 , 2013
Abstract : Monoacylglycerol lipase (MAGL) is a principal metabolic enzyme responsible for hydrolyzing the endogenous cannabinoid (endocannabinoid) 2-arachidonoylglycerol (2-AG). Selective inhibitors of MAGL offer valuable probes to further understand the enzyme's function in biological systems and may lead to drugs for treating a variety of diseases, including psychiatric disorders, neuroinflammation, and pain. N-Hydroxysuccinimidyl (NHS) carbamates have recently been identified as a promising class of serine hydrolase inhibitors that shows minimal cross-reactivity with other proteins in the proteome. Here, we explore NHS carbamates more broadly and demonstrate their potential as inhibitors of endocannabinoid hydrolases and additional enzymes from the serine hydrolase class. We extensively characterize an NHS carbamate 1a (MJN110) as a potent, selective, and in-vivo-active MAGL inhibitor. Finally, we demonstrate that MJN110 alleviates mechanical allodynia in a rat model of diabetic neuropathy, marking NHS carbamates as a promising class of MAGL inhibitors.
ESTHER : Niphakis_2013_ACS.Chem.Neurosci_4_1322
PubMedSearch : Niphakis_2013_ACS.Chem.Neurosci_4_1322
PubMedID: 23731016
Gene_locus related to this paper: human-MGLL

Title : Dual inhibition of endocannabinoid catabolic enzymes produces enhanced antiwithdrawal effects in morphine-dependent mice - Ramesh_2013_Neuropsychopharmacology_38_1039
Author(s) : Ramesh D , Gamage TF , Vanuytsel T , Owens RA , Abdullah RA , Niphakis MJ , Shea-Donohue T , Cravatt BF , Lichtman AH
Ref : Neuropsychopharmacology , 38 :1039 , 2013
Abstract : Inhibition of the endocannabinoid catabolic enzymes, monoacylglycerol lipase (MAGL) or fatty acid amide hydrolase (FAAH) attenuates naloxone-precipitated opioid withdrawal signs in mice via activation of CB1 receptors. Complete FAAH inhibition blocks only a subset of withdrawal signs, whereas complete MAGL inhibition elicits enhanced antiwithdrawal efficacy, but is accompanied with some cannabimimetic side effects. Thus, the primary objective of the present study was to determine whether combined, full FAAH inhibition and partial MAGL represents an optimal strategy to reduce opioid withdrawal. To test this hypothesis, we examined whether combined administration of high-dose of the FAAH inhibitor PF-3845 and low-dose of the MAGL inhibitor JZL184, as well as the novel dual FAAH-MAGL inhibitor SA-57, which is 100-fold more potent in inhibiting FAAH than MAGL, would prevent spontaneous withdrawal in morphine-dependent mice, a model with greater face validity than precipitating withdrawal with mu-opioid receptor antagonists. Strikingly, a combination of low-dose JZL184 and high-dose PF-3845 as well as the dual inhibitor SA-57 reduced all abrupt withdrawal signs (ie, platform jumping, paw flutters, head shakes, diarrhea, and total body weight loss), but did not elicit any cannabimimetic side effects. In addition, JZL184 or PF-3845 blocked naloxone-precipitated hypersecretion in morphine-dependent small intestinal tissue. Collectively, these results are the first to show that endocannabinoid catabolic enzyme inhibitors reduce abrupt withdrawal in morpine-dependent mice and are effective in a novel in vitro model of opioid withdrawal. More generally, these findings support the idea that joint MAGL and FAAH inhibition represents a promising approach for the treatment of opioid dependence.
ESTHER : Ramesh_2013_Neuropsychopharmacology_38_1039
PubMedSearch : Ramesh_2013_Neuropsychopharmacology_38_1039
PubMedID: 23303065
Gene_locus related to this paper: human-MGLL

Title : O-hydroxyacetamide carbamates as a highly potent and selective class of endocannabinoid hydrolase inhibitors - Niphakis_2012_ACS.Chem.Neurosci_3_418
Author(s) : Niphakis MJ , Johnson DS , Ballard TE , Stiff C , Cravatt BF
Ref : ACS Chem Neurosci , 3 :418 , 2012
Abstract : The two major endocannabinoid transmitters, anandamide (AEA) and 2-arachidonoylglycerol (2-AG), are degraded by distinct enzymes in the nervous system, fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL), respectively. FAAH and MAGL inhibitors cause elevations in brain AEA and 2-AG levels, respectively, and reduce pain, anxiety, and depression in rodents without causing the full spectrum of psychotropic behavioral effects observed with direct cannabinoid receptor-1 (CB1) agonists. These findings have inspired the development of several classes of endocannabinoid hydrolase inhibitors, most of which have been optimized to show specificity for either FAAH or MAGL or, in certain cases, equipotent activity for both enzymes. Here, we investigate an unusual class of O-hydroxyacetamide carbamate inhibitors and find that individual compounds from this class can serve as selective FAAH or dual FAAH/MAGL inhibitors in vivo across a dose range (0.125-12.5 mg kg(-1)) suitable for behavioral studies. Competitive and click chemistry activity-based protein profiling confirmed that the O-hydroxyacetamide carbamate SA-57 is remarkably selective for FAAH and MAGL in vivo, targeting only one other enzyme in brain, the additional 2-AG hydrolase ABHD6. These data designate O-hydroxyacetamide carbamates as a versatile chemotype for creating endocannabinoid hydrolase inhibitors that display excellent in vivo activity and tunable selectivity for FAAH-anandamide versus MAGL (and ABHD6)-2-AG pathways.
ESTHER : Niphakis_2012_ACS.Chem.Neurosci_3_418
PubMedSearch : Niphakis_2012_ACS.Chem.Neurosci_3_418
PubMedID: 22860211
Gene_locus related to this paper: human-MGLL

Title : Highly selective inhibitors of monoacylglycerol lipase bearing a reactive group that is bioisosteric with endocannabinoid substrates - Chang_2012_Chem.Biol_19_579
Author(s) : Chang JW , Niphakis MJ , Lum KM , Cognetta AB, 3rd , Wang C , Matthews ML , Niessen S , Buczynski MW , Parsons LH , Cravatt BF
Ref : Chemical Biology , 19 :579 , 2012
Abstract : The endocannabinoids 2-arachidonoyl glycerol (2-AG) and N-arachidonoyl ethanolamine (anandamide) are principally degraded by monoacylglycerol lipase (MAGL) and fatty acid amide hydrolase (FAAH), respectively. The recent discovery of O-aryl carbamates such as JZL184 as selective MAGL inhibitors has enabled functional investigation of 2-AG signaling pathways in vivo. Nonetheless, JZL184 and other reported MAGL inhibitors still display low-level cross-reactivity with FAAH and peripheral carboxylesterases, which can complicate their use in certain biological studies. Here, we report a distinct class of O-hexafluoroisopropyl (HFIP) carbamates that inhibits MAGL in vitro and in vivo with excellent potency and greatly improved selectivity, including showing no detectable cross-reactivity with FAAH. These findings designate HFIP carbamates as a versatile chemotype for inhibiting MAGL and should encourage the pursuit of other serine hydrolase inhibitors that bear reactive groups resembling the structures of natural substrates.
ESTHER : Chang_2012_Chem.Biol_19_579
PubMedSearch : Chang_2012_Chem.Biol_19_579
PubMedID: 22542104
Gene_locus related to this paper: human-MGLL