Nevalainen T

References (11)

Title : In Vivo Characterization of the Ultrapotent Monoacylglycerol Lipase Inhibitor {4-[bis-(benzo[d][1,3]dioxol-5-yl)methyl]-piperidin-1-yl}(1H-1,2,4-triazol-1-yl)m ethanone (JJKK-048) - Aaltonen_2016_J.Pharmacol.Exp.Ther_359_62
Author(s) : Aaltonen N , Kedzierska E , Orzelska-Gorka J , Lehtonen M , Navia-Paldanius D , Jakupovic H , Savinainen JR , Nevalainen T , Laitinen JT , Parkkari T , Gynther M
Ref : Journal of Pharmacology & Experimental Therapeutics , 359 :62 , 2016
Abstract : Monoacylglycerol lipase (MAGL) is a serine hydrolase that acts as a principal degradative enzyme for the endocannabinoid 2-arachidonoylglycerol (2-AG). In addition to terminating the signaling function of 2-AG, MAGL liberates arachidonic acid to be used as a primary source for neuroinflammatory prostaglandin synthesis in the brain. MAGL activity also contributes to cancer pathogenicity by producing precursors for tumor-promoting bioactive lipids. Pharmacological inhibitors of MAGL provide valuable tools for characterization of MAGL and 2-AG signaling pathways. They also hold great therapeutic potential to treat several pathophysiological conditions, such as pain, neurodegenerative disorders, and cancer. We have previously reported piperidine triazole urea, {4-[bis-(benzo[d][1,3]dioxol-5-yl)methyl]-piperidin-1-yl}(1H-1,2,4-triazol-1-yl)m ethanone (JJKK-048), to be an ultrapotent and highly selective inhibitor of MAGL in vitro. Here, we characterize in vivo effects of JJKK-048. Acute in vivo administration of JJKK-048 induced a massive increase in mouse brain 2-AG levels without affecting brain anandamide levels. JJKK-048 appeared to be extremely potent in vivo. Activity-based protein profiling revealed that JJKK-048 maintains good selectivity toward MAGL over other serine hydrolases. Our results are also the first to show that JJKK-048 promoted significant analgesia in a writhing test with a low dose that did not cause cannabimimetic side effects. At a high dose, JJKK-048 induced analgesia both in the writhing test and in the tail-immersion test, as well as hypomotility and hyperthermia, but not catalepsy.
ESTHER : Aaltonen_2016_J.Pharmacol.Exp.Ther_359_62
PubMedSearch : Aaltonen_2016_J.Pharmacol.Exp.Ther_359_62
PubMedID: 27451409

Title : Discovery of Triterpenoids as Reversible Inhibitors of alpha\/beta-hydrolase Domain Containing 12 (ABHD12) - Parkkari_2014_PLoS.One_9_e98286
Author(s) : Parkkari T , Haavikko R , Laitinen T , Navia-Paldanius D , Rytilahti R , Vaara M , Lehtonen M , Alakurtti S , Yli-Kauhaluoma J , Nevalainen T , Savinainen JR , Laitinen JT
Ref : PLoS ONE , 9 :e98286 , 2014
Abstract : BACKGROUND: alpha/beta-hydrolase domain containing (ABHD)12 is a recently discovered serine hydrolase that acts in vivo as a lysophospholipase for lysophosphatidylserine. Dysfunctional ABHD12 has been linked to the rare neurodegenerative disorder called PHARC (polyneuropathy, hearing loss, ataxia, retinosis pigmentosa, cataract). In vitro, ABHD12 has been implicated in the metabolism of the endocannabinoid 2-arachidonoylglycerol (2-AG). Further studies on ABHD12 function are hampered as no selective inhibitor have been identified to date. In contrast to the situation with the other endocannabinoid hydrolases, ABHD12 has remained a challenging target for inhibitor development as no crystal structures are available to facilitate drug design. METHODOLOGY/PRINCIPAL FINDINGS: Here we report the unexpected discovery that certain triterpene-based structures inhibit human ABHD12 hydrolase activity in a reversible manner, the best compounds showing submicromolar potency. Based on structure activity relationship (SAR) data collected for 68 natural and synthetic triterpenoid structures, a pharmacophore model has been constructed. A pentacyclic triterpene backbone with carboxyl group at position 17, small hydrophobic substituent at the position 4, hydrogen bond donor or acceptor at position 3 accompanied with four axial methyl substituents was found crucial for ABHD12 inhibitor activity. Although the triterpenoids typically may have multiple protein targets, we witnessed unprecedented selectivity for ABHD12 among the metabolic serine hydrolases, as activity-based protein profiling of mouse brain membrane proteome indicated that the representative ABHD12 inhibitors did not inhibit other serine hydrolases, nor did they target cannabinoid receptors. CONCLUSIONS/SIGNIFICANCE: We have identified reversibly-acting triterpene-based inhibitors that show remarkable selectivity for ABHD12 over other metabolic serine hydrolases. Based on SAR data, we have constructed the first pharmacophore model of ABHD12 inhibitors. This model should pave the way for further discovery of novel lead structures for ABHD12 selective inhibitors.
ESTHER : Parkkari_2014_PLoS.One_9_e98286
PubMedSearch : Parkkari_2014_PLoS.One_9_e98286
PubMedID: 24879289
Gene_locus related to this paper: human-ABHD12

Title : Biochemical and Pharmacological Characterization of the Human Lymphocyte Antigen B-Associated Transcript 5 (BAT5\/ABHD16A) - Savinainen_2014_PLoS.One_9_e109869
Author(s) : Savinainen JR , Patel JZ , Parkkari T , Navia-Paldanius D , Marjamaa JJ , Laitinen T , Nevalainen T , Laitinen JT
Ref : PLoS ONE , 9 :e109869 , 2014
Abstract : BACKGROUND: Human lymphocyte antigen B-associated transcript 5 (BAT5, also known as ABHD16A) is a poorly characterized 63 kDa protein belonging to the alpha/beta-hydrolase domain (ABHD) containing family of metabolic serine hydrolases. Its natural substrates and biochemical properties are unknown. METHODOLOGY/PRINCIPAL FINDINGS: Amino acid sequence comparison between seven mammalian BAT5 orthologs revealed that the overall primary structure was highly (>/=95%) conserved. Activity-based protein profiling (ABPP) confirmed successful generation of catalytically active human (h) and mouse (m) BAT5 in HEK293 cells, enabling further biochemical characterization. A sensitive fluorescent glycerol assay reported hBAT5-mediated hydrolysis of medium-chain saturated (C14ratio0), long-chain unsaturated (C18ratio1, C18ratio2, C20ratio4) monoacylglycerols (MAGs) and 15-deoxy-Delta12,14-prostaglandin J2-2-glycerol ester (15d-PGJ2-G). In contrast, hBAT5 possessed only marginal diacylglycerol (DAG), triacylglycerol (TAG), or lysophospholipase activity. The best MAG substrates were 1-linoleylglycerol (1-LG) and 15d-PGJ2-G, both exhibiting low-micromolar Km values. BAT5 had a neutral pH optimum and showed preference for the 1(3)- vs. 2-isomers of MAGs C18ratio1, C18ratio2 and C20ratio4. Inhibitor profiling revealed that beta-lactone-based lipase inhibitors were nanomolar inhibitors of hBAT5 activity (palmostatin B > tetrahydrolipstatin > ebelactone A). Moreover, the hormone-sensitive lipase inhibitor C7600 (5-methoxy-3-(4-phenoxyphenyl)-3H-[1], [3], [4]oxadiazol-2-one) was identified as a highly potent inhibitor (IC50 8.3 nM). Phenyl and benzyl substituted analogs of C7600 with increased BAT5 selectivity were synthesized and a preliminary SAR analysis was conducted to obtain initial insights into the active site dimensions. CONCLUSIONS/SIGNIFICANCE: This study provides an initial characterization of BAT5 activity, unveiling the biochemical and pharmacological properties with in vitro substrate preferences and inhibitor profiles. Utilization of glycerolipid substrates and sensitivity to lipase inhibitors suggest that BAT5 is a genuine lipase with preference for long-chain unsaturated MAGs and could in this capacity regulate glycerolipid metabolism in vivo as well. This preliminary SAR data should pave the way towards increasingly potent and BAT5-selective inhibitors.
ESTHER : Savinainen_2014_PLoS.One_9_e109869
PubMedSearch : Savinainen_2014_PLoS.One_9_e109869
PubMedID: 25290914
Gene_locus related to this paper: human-ABHD16A , mouse-Abhd16a

Title : Robust Hydrolysis of Prostaglandin Glycerol Esters by Human Monoacylglycerol Lipase (MAGL) - Savinainen_2014_Mol.Pharmacol_86_522
Author(s) : Savinainen JR , Kansanen E , Pantsar T , Navia-Paldanius D , Parkkari T , Lehtonen M , Laitinen T , Nevalainen T , Poso A , Levonen AL , Laitinen JT
Ref : Molecular Pharmacology , 86 :522 , 2014
Abstract : The primary route of inactivation of the endocannabinoid 2-arachidonoylglycerol in the central nervous system is through enzymatic hydrolysis, mainly carried out by monoacylglycerol lipase (MAGL), along with a small contribution by the alpha/beta-hydrolase domain (ABHD) proteins ABHD6 and ABHD12. Recent methodological progress allowing kinetic monitoring of glycerol liberation has facilitated substrate profiling of the human endocannabinoid hydrolases, and these studies have revealed that the three enzymes have distinct monoacylglycerol substrate and isomer preferences. Here, we have extended this substrate profiling to cover four prostaglandin glycerol esters, namely, 15-deoxy-Delta(12,14)-prostaglandin J2-2-glycerol (15d-PGJ2-G), PGD2-G, PGE2-G, and PGF2 alpha-G. We found that the three enzymes hydrolyzed the tested substrates, albeit with distinct rates and preferences. Although human ABHD12 (hABHD12) showed only marginal activity toward PGE2-G, hABHD6 preferentially hydrolyzed PGD2-G, and human MAGL (hMAGL) robustly hydrolyzed all four. This was particularly intriguing for MAGL activity toward 15d-PGJ2-G whose hydrolysis rate rivaled that of the best monoacylglycerol substrates. Molecular modeling studies combined with kinetic analysis supported favorable interaction with the hMAGL active site. Long and short MAGL isoforms shared a similar substrate profile, and hMAGL hydrolyzed 15d-PGJ2-G also in living cells. The ability of 15d-PGJ2-G to activate the canonical nuclear factor erythroid 2-related factor (Nrf2) signaling pathway used by 15d-PGJ2 was assessed, and these studies revealed for the first time that 15d-PGJ2 and 15d-PGJ2-G similarly activated Nrf2 signaling as well as transcription of target genes of this pathway. Our study challenges previous claims regarding the ability of MAGL to catalyze PG-G hydrolysis and extend the MAGL substrate profile beyond the classic monoacylglycerols.
ESTHER : Savinainen_2014_Mol.Pharmacol_86_522
PubMedSearch : Savinainen_2014_Mol.Pharmacol_86_522
PubMedID: 25140003
Gene_locus related to this paper: human-ABHD6 , human-ABHD12 , human-MGLL

Title : Piperazine and piperidine triazole ureas as ultrapotent and highly selective inhibitors of monoacylglycerol lipase - Aaltonen_2013_Chem.Biol_20_379
Author(s) : Aaltonen N , Savinainen JR , Ribas CR , Ronkko J , Kuusisto A , Korhonen J , Navia-Paldanius D , Hayrinen J , Takabe P , Kasnanen H , Pantsar T , Laitinen T , Lehtonen M , Pasonen-Seppanen S , Poso A , Nevalainen T , Laitinen JT
Ref : Chemical Biology , 20 :379 , 2013
Abstract : Monoacylglycerol lipase (MAGL) terminates the signaling function of the endocannabinoid, 2-arachidonoylglycerol (2-AG). During 2-AG hydrolysis, MAGL liberates arachidonic acid, feeding the principal substrate for the neuroinflammatory prostaglandins. In cancer cells, MAGL redirects lipid stores toward protumorigenic signaling lipids. Thus MAGL inhibitors may have great therapeutic potential. Although potent and increasingly selective MAGL inhibitors have been described, their number is still limited. Here, we have characterized piperazine and piperidine triazole ureas that combine the high potency attributable to the triazole leaving group together with the bulky aromatic benzodioxolyl moiety required for selectivity, culminating in compound JJKK-048 that potently (IC50 < 0.4 nM) inhibited human and rodent MAGL. JJKK-048 displayed low cross-reactivity with other endocannabinoid targets. Activity-based protein profiling of mouse brain and human melanoma cell proteomes suggested high specificity also among the metabolic serine hydrolases.
ESTHER : Aaltonen_2013_Chem.Biol_20_379
PubMedSearch : Aaltonen_2013_Chem.Biol_20_379
PubMedID: 23521796
Gene_locus related to this paper: human-MGLL

Title : 1,3,4-Oxadiazol-2-ones as fatty-acid amide hydrolase and monoacylglycerol lipase inhibitors: Synthesis, in vitro evaluation and insight into potency and selectivity determinants by molecular modelling - Kasnanen_2013_Eur.J.Pharm.Sci_49_423
Author(s) : Kasnanen H , Minkkila A , Taupila S , Patel JZ , Parkkari T , Lahtela-Kakkonen M , Saario SM , Nevalainen T , Poso A
Ref : Eur J Pharm Sci , 49 :423 , 2013
Abstract : Inhibition of the key hydrolytic enzymes of the endocannabinoid system, fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL), has been proposed as potential mode of action for various therapeutic applications. Continuing our previous work, we take the first steps of structure-activity relationship exploration and show that 1,3,4-oxadiazol-2-ones can serve as scaffold for both selective FAAH and MAGL inhibitors, and also function as a dual FAAH/MAGL inhibitor at sub-micromolar IC50 values. Moreover, 10-fold selectivity against MAGL over FAAH was achieved with compound 3d (FAAH and MAGL IC50; 2.0 and 0.22muM). Lastly, enzyme and ligand features contributing to the potency and selectivity differences are analysed by molecular docking.
ESTHER : Kasnanen_2013_Eur.J.Pharm.Sci_49_423
PubMedSearch : Kasnanen_2013_Eur.J.Pharm.Sci_49_423
PubMedID: 23557840
Gene_locus related to this paper: human-MGLL

Title : Discovery and development of endocannabinoid-hydrolyzing enzyme inhibitors - Minkkila_2010_Curr.Top.Med.Chem_10_828
Author(s) : Minkkila A , Saario SM , Nevalainen T
Ref : Curr Top Med Chem , 10 :828 , 2010
Abstract : Fatty acid amide hydrolase (FAAH) and monoglyceride lipase (MGL) are hydrolytic enzymes which degrade the endogenous cannabinoids (endocannabinoids) N-arachidonoylethanolamine (anandamide, AEA) and 2-arachidonoylglycerol (2-AG), respectively. Endocannabinoids are an important class of lipid messenger molecules that are produced on demand in response to elevated intracellular calcium levels. They recognize and activate the cannabinoid CB(1) and CB(2) receptors, the molecular targets for Delta(9)-tetrahydrocannabinol (Delta(9)-THC) in marijuana evoking several beneficial therapeutic effects. However, in vivo the cannabimimetic effects of AEA and 2-AG remain weak owing to their rapid inactivation by FAAH and MGL, respectively. The inactivation of FAAH and MGL by specific enzyme inhibitors increases the levels of AEA and 2-AG, respectively, producing therapeutic effects such as pain relief and depression of anxiety.
ESTHER : Minkkila_2010_Curr.Top.Med.Chem_10_828
PubMedSearch : Minkkila_2010_Curr.Top.Med.Chem_10_828
PubMedID: 20370710

Title : Characterization of binding properties of monoglyceride lipase inhibitors by a versatile fluorescence-based technique - Savinainen_2010_Anal.Biochem_399_132
Author(s) : Savinainen JR , Yoshino M , Minkkila A , Nevalainen T , Laitinen JT
Ref : Analytical Biochemistry , 399 :132 , 2010
Abstract : Monoglyceride lipase (MGL) is a serine hydrolase that terminates the signaling of the primary endocannabinoid, 2-arachidonoyl glycerol (2-AG). Versatile high-throughput screening methods allowing the testing of MGL inhibitors are rare, thereby limiting the development and analysis of novel inhibitors. Here we describe an improved fluorescence-based technique that is capable of determining time- and dose-dependent inhibition of MGL with one or multiple binding sites and, at the same time, is capable of revealing the reversibility of inhibitor binding in a simple kinetic assay format. Known reference compounds as well as novel inhibitors, such as JZL184 and CAY10499, were evaluated for their MGL-binding properties and potency.
ESTHER : Savinainen_2010_Anal.Biochem_399_132
PubMedSearch : Savinainen_2010_Anal.Biochem_399_132
PubMedID: 20005861
Gene_locus related to this paper: human-MGLL

Title : Screening of various hormone-sensitive lipase inhibitors as endocannabinoid-hydrolyzing enzyme inhibitors -
Author(s) : Minkkila A , Savinainen JR , Kasnanen H , Xhaard H , Nevalainen T , Laitinen JT , Poso A , Leppanen J , Saario SM
Ref : ChemMedChem , 4 :1253 , 2009
PubMedID: 19472270

Title : Design, synthesis, and in vitro evaluation of carbamate derivatives of 2-benzoxazolyl- and 2-benzothiazolyl-(3-hydroxyphenyl)-methanones as novel fatty acid amide hydrolase inhibitors - Myllymaki_2007_J.Med.Chem_50_4236
Author(s) : Myllymaki MJ , Saario SM , Kataja AO , Castillo-Melendez JA , Nevalainen T , Juvonen RO , Jarvinen T , Koskinen AM
Ref : Journal of Medicinal Chemistry , 50 :4236 , 2007
Abstract : Fatty acid amide hydrolase (FAAH) is an intracellular serine hydrolase, which catalyzes the hydrolysis of the endocannabinoid N-arachidonoylethanolamide to arachidonic acid and ethanolamine. FAAH also hydrolyzes another endocannabinoid, 2-arachidonoylglycerol (2-AG). However, 2-AG has been assumed to be hydrolyzed mainly by monoacylglycerol lipase (MAGL) or a MAGL-like enzyme. Inhibition of FAAH or MAGL activity might lead to beneficial effects in many physiological disorders such as pain, inflammation, and anxiety due to increased endocannabinoid-induced activation of cannabinoid receptors CB1 and CB2. In the present study, a total of 34 novel compounds were designed, synthesized, characterized, and tested against FAAH and MAGL-like enzyme activity. Altogether, 16 compounds were found to inhibit FAAH with half-maximal inhibition concentrations (IC50) between 28 and 380 nM. All the active compounds belong to the structural family of carbamates. Compounds 14 and 18 were found to be the most potent FAAH inhibitors, which may serve as lead structures for novel FAAH inhibitors.
ESTHER : Myllymaki_2007_J.Med.Chem_50_4236
PubMedSearch : Myllymaki_2007_J.Med.Chem_50_4236
PubMedID: 17665899

Title : Characterization of the sulfhydryl-sensitive site in the enzyme responsible for hydrolysis of 2-arachidonoyl-glycerol in rat cerebellar membranes - Saario_2005_Chem.Biol_12_649
Author(s) : Saario SM , Salo OM , Nevalainen T , Poso A , Laitinen JT , Jarvinen T , Niemi R
Ref : Chemical Biology , 12 :649 , 2005
Abstract : We have previously reported that the endocannabinoid, 2-arachidonoyl-glycerol (2-AG), is hydrolyzed in rat cerebellar membranes by monoglyceride lipase (MGL)-like enzymatic activity. The present study shows that, like MGL, 2-AG-degrading enzymatic activity is sensitive to inhibition by sulfhydryl-specific reagents. Inhibition studies of this enzymatic activity by N-ethylmaleimide analogs revealed that analogs with bulky hydrophobic N-substitution were more potent inhibitors than hydrophilic or less bulky agents. Interestingly, the substrate analog N-arachidonylmaleimide was found to be the most potent inhibitor. A comparison model of MGL was constructed to get a view on the cysteine residues located near the binding site. These findings support our previous conclusion that the 2-AG-degrading enzymatic activity in rat cerebellar membranes corresponds to MGL or MGL-like enzyme and should facilitate further efforts to develop potent and more selective MGL inhibitors.
ESTHER : Saario_2005_Chem.Biol_12_649
PubMedSearch : Saario_2005_Chem.Biol_12_649
PubMedID: 15975510