Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. or Hipp Joerg Novel a GABA Basmisanil, Selective of Pharmacology Clinical and Preclinical ehr Trube Gerhard oof Gasser Rodolfo Hipp F Joerg authors all of names Full GABA selective highly a Basmisanil, title GABA running Selective Short Novel a Basmisanil, of Modulator losteric Pharmacology Clinical and Preclinical Title disorders. GABA brain selective multiple and in potent testing highly clinical a for allowing is There tolerability Basmisanil power. and Implications safety spectral and receptor EEG good to Conclusion of with changes exposure events. NAM characteristic plasma adverse in the serious reflected and established anxiogenic no function and no brain were engagement had modulated target basmisanil Basmisanil showed concentrations, plasma relationship. PET efficacious volunteers, occupancy these healthy At In NHPs. impairment effects. in learning proconvulsant function spatial executive diazepam-induced improved attenuated and Basmisanil rats rats. in in engagement target dose-dependent demonstrated GABA GABA human at recombinant currents to bound GABA-induced modulation Basmisanil and inhibited Results engagement Key target EEG. and volunteers, and retrieval) healthy PET object In using α (NHP; assessed rats. GABA primates were in target activity non-human of tested network and measured and were neuronal maze) (NAM) studies proconvulsant) of preclinical water and occupancy (Morris modulator the (anxiety vivo rats effects describe allosteric In in side We selectivity. tested negative potential were functional selective cognition disorders. and on other binding highly Effects assessed as and engagement. assays well potent vitro as In a dysfunction, Approach basmisanil, Experimental cognitive for of target profile drug clinical promising a represent and GABA Purpose and Background Abstract 2020 16, July 1 Noeldeke Jana Knust Henner eetr ih5n ffiiyadmr hn9-odslciiyversus selectivity 90-fold than more and affinity nM 5 with receptors 5 omn-aRceA eerhadDvlpetDivision Development and Research AG Roche Hoffmann-La F 1 1 rdrcKnoflach Frederic , Fr´ed´eric, Knoflach 2 1 rcPrinssen Eric , 1 inLennon-Chrimes Sian , 1 hitp Wandel Christoph , oof Gasser Rodolfo , A A - α - α uui-otiigrcposhv ensont lyakymdltr oei cognition in role modulatory key a play to shown been have receptors subunit-containing 5 eetrNgtv lotrcModulator Allosteric Negative Receptor 5 1 1 ay .Wallace L. Tanya , oetComley Robert , 1 A oetComley Robert , A 1 - - α rcP Prinssen P. Eric , α NAM 5 e a iteo oeeta h te eetrsbye.I io basmisanil vivo, In subtypes. receptor other the at effect no or little had yet 5 1 nrwThomas Andrew , 1 ihe Derks Michael , 1 hrs .Ballard M. Theresa , 5 1 nra Rothfuss Andreas , 1 hrs Ballard Theresa , 1 α ay Wallace Tanya , 1, 1 α 1 n ai-lmni Hernandez Maria-Clemencia and , ,and 2, iaSquassante Lisa , α uui-otiigrcpos Basmisanil receptors. subunit-containing 3 1 ihe Honer Michael , 2 enrKnust Henner , 1 A 1 ihe Honer Michael , nra Rothfuss Andreas , - α eetrNgtv Al- Negative Receptor 5 1 tpaeNave Stephane , 1 ehr Trube Gerhard , 3 inLennon- Sian , A A α receptors. 5 α 1 receptor 5 , 1 , 1 A 1 1 , - , Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. l uhr eeepoeso .omn-aRceA wteln rRceBocec S rRoche or studies. USA clinical Bioscience and preclinical Roche the or of Switzerland time AG the Roche at F.Hoffmann-La UK Ltd. of Products employees were authors All Ltd.). E. statement Imanova Graham study, interest Brown, (PET P. of Rabiner Andrew Conflicts A. pharmacology), Eugenii (clinical product), Gunn, Bentley (drug N. Wyttenbach Darren Roger Nicole and Searle, (chemistry), Diack Waldmeier Cheikh Theo Pius Jamois, Schaer, Husar, Roger Heidi Candice and Kurt, Elisabeth Anke Kahn development), Koblet, Francoise Andreas (neuroscience Haman, Bj Seneca Marie Greiter-Wilke, (molecular Andrea Dao, Nicholas Dinklo, Karg Hung pharmacology), Maria Chiu, ( (behavioural Gabriel and Graf Wyler Bandinelli, Veronique Harle-Yge Severine and Marie-Laurence Algeyer, Pflimlin Guizani, Brigitte Pascal Cecile assays), Friz, binding Gregoire biology, thank authors The Acknowledgements 250 Abstract: 1498 Conclusions: & Discussion fi- and references methods, legends): abstract, gure (excluding 3996 count: Word USA. Illinois, Chicago, North Inc, AbbVie RC: Switzerland. Zurich, AG, Capital Helvetica UK. AWT: Devon, USA Dartmouth, 94103, Ltd, California (UK) Francisco, Galwyn San TMB: Street, Brannan 780 Therapeutics, BlackThorn TLW: Affiliation Current UK. 1TW, 5 AL7 City, Garden Welwyn wyn, Grenzacherstrasse 4 Basel, Center Switzerland. Innovation Basel, Roche 4070 Development, 124, Early and Research Pharmaceutical Roche 3 2 out) carried was work (where 1 affiliations institutional authors All Thomas W Chrimes hraetclSine,RcePam eerhadEryDvlpet oh noainCne Wel- Center Innovation Roche Development, Early and Research Pharma Roche Sciences, Pharmaceutical N eerh oh isine aoAt,USA Alto, Palo Bioscience, Roche Research, CNS Research Molecule Small Area Translational Sciences and Pharmaceutical Discovery (NRD) Diseases Rare & Neuroscience • • eut:1838 Results: 660 Introduction: 4 ihe Derks Michael , 3 ai-lmni Hernandez Maria-Clemencia , 4 iaSquassante Lisa , r aosn oiu cmt n rc oz(hraetclsciences), (pharmaceutical Wolz Erich and Schmitt Monique Jacobsen, ¨ orn 1 4 tpaeNave Stephane , 2 1 aaN Jana , Xenopus ¨ oldeke oyei-ir electrophysiology), in-vitro oocyte 1 hitp Wandel Christoph , 2 Andrew , Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. GABA Purpose and Background Abstract rciia n lnclpol fbsiai,aptn n ihyslciengtv lotrcmodulator allosteric the negative describe selective We highly disorders. and other potent as a well basmisanil, as of GABA dysfunction, of profile cognitive (NAM) for clinical target and drug preclinical promising a represent GABA versus GABA lectivity human healthy recombinant In to bound and Basmisanil rats. PET using in assessed tested were Results target were activity Key measured network proconvulsant) (NHP; neuronal studies and primates of non-human (anxiety occupancy modulation and effects EEG. vivo maze) and water side engagement In (Morris target potential rats volunteers, selectivity. in and tested functional retrieval) were and object cognition on binding Effects assessed engagement. assays vitro In Approach Experimental lnclsignificance Clinical GABA selective and potent highly a events. is adverse serious Basmisanil no function were and brain There engagement Implications modulated power. target and Basmisanil spectral showed basmisanil Conclusion EEG PET relationship. concentrations, of volunteers, occupancy plasma changes healthy receptor characteristic efficacious in to In these reflected exposure At effects. plasma proconvulsant the NHPs. and established in anxiogenic impairment function learning no executive spatial improved had diazepam-induced and attenuated Basmisanil rats rats. in in engagement target dependent htti td adds study GABA this of What importance the cognition. demonstrated on have receptors studies pharmacological and Genetic known already is What summary point Bullet disorders. brain multiple in testing clinical for allowing • • • • • amsnlhsa da rfiet netgt oeta lnclbnfiso GABA of benefits clinical potential investigate to modulation. profile GABA ideal maximum an at has humans Basmisanil in tolerability and safety good shows Basmisanil effects pharmacodynamic and (EEG). humans) clinically (rats, and engagement (cognition) preclinically target dose-dependent showed Basmisanil far. so amsnli h otptn n ihyslcieGABA selective highly and potent most the is Basmisanil A A - - α α uui-otiigrcposhv ensont lyakymdltr oei onto and cognition in role modulatory key a play to shown been have receptors subunit-containing 5 e a iteo oeeta h te eetrsbye.I io amsnldmntae dose- demonstrated basmisanil vivo, In subtypes. receptor other the at effect no or little had yet 5 α 1, A - α α ,and 2, receptors. 5 α uui-otiigrcpos amsnlihbtdGB-nue urnsat currents GABA-induced inhibited Basmisanil receptors. subunit-containing 3 A - α eetr ih5n ffiiyadmr hn9-odse- 90-fold than more and affinity nM 5 with receptors 5 A 3 - α eetrNMwt odsft n tolerability and safety good with NAM receptor 5 A - α eaiealsei ouao described modulator allosteric negative 5 A A - α - A α eetroccupancy. receptor 5 - subunit-containing 5 α eetrnegative receptor 5 Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. ltc,sdtvs yntc rateietc idt h lotrcbnoizpn BD idn iewhich site binding the (BZD) of benzodiazepine one allosteric the the by to of formed bind interface is antiepileptics or the hypnotics at sedatives, sites olytics, binding two has GABA Basmisanil, Keywords soitdwt ontv maret oee,i a o enpsil ociial sesteeet of effects the assess clinically to possible been not has GABA it at 2017). However, activity al., reduced impairment. et Zanos cognitive 2015; with al., GABA associated Recently, et (Fischell 2012). GABA mice mice al., in Therefore, in et effects studies antidepressant-like Schmidt rapid 2015; addition, GABA exert al., GABA In to of et increased shown Lake reduction Stamenic 2014). is 2010; that Timic there al., etal., stroke and 2012; et (Clarkson ischemic area al., (Zurek after et anesthesia peri-infarct that after Redrobe the shown impairment 2014; have improvements cognitive etal., rats by and (Povroznik and shown 2015) dysfunction as NMDA schizophrenia al., with of et GABA associated models that impairment preclinical suggest cognitive in studies 2013), preclinical al., further et then, Martinez-Cue Since 2003). GABA α (Maubach, selective al., impairment et that cognitive (Kawaharada mild proposed ONO-8290580 was 2009), it al., Initially, et (Ballard non-selective RO4938581 with 2008), as, such al., associated 2018). 1984), et effects etal., been (Savic side Little have PWZ-029 proconvulsant 1983; compounds 2009), al., or Several et anxiogenic (Dorow 2011). expres- the NAMs Knoflach, preferential without & their the enhancement (Rudolph with for cognitive line regions selectivity in cortical possess processes, and to memory described hippocampus and the learning in in GABA role sion that modulatory demonstrated important have an studies play GABA pharmacological of and activity genetic today. the Both developed decrease of which successfully activity ligands been the site not increase BZD-binding have but are receptors, alone, (NAMs) effect modulators no allosteric have negative they since (PAMs) modulators GABA allosteric positive as known two of stoichiometry GABA common for most encoding the genes with 19 are There GABA. transmitter, GABA Introduction 1 uonuto rprisi aslaigt eue lsaepsr ncrncdsn Bngade al., et (Bundgaard dosing chronic on GABA CYP1A2 exposure strong for of plasma search because species reduced continued abandoned our preclinical to be but leading in to had toxicity 2013), rats pharmacokinetics, RO4938581, renal variable in as 2009). of properties well al., as autoinduction because humans et elderly Atack stopped healthy 2010; in was (Atack, tolerability development respectively poor and their 2005) al., but et trials, (Merschman clinical in tested xoueadrcpo cuac nrt i ioatrdorpy n uas(E) 2.cluaeesti- calculate (2). (PET); plasma humans between and relationship autoradiography) the establish vivo (in (1). rats to: basmisanil in of occupancy effects the receptor assess and to was exposure study this of aim The hits. [ initial on GABA the cloned of based activity on screen experiments functional electrophysiological high-throughput by followed library binding compound molecular-weight small ( iue1 Figure Asmyatnaecgiieipimn soitdwt onsnrm Bada ta. 2011; al., et (Braudeau syndrome Down with associated impairment cognitive attenuate may NAMs 5 A A eetr ntepeec fGB Sehr ai,21;Sgl&Senan 02.I contrast, In 2012). Steinmann, & Sigel 2018; Savic, & (Sieghart GABA of presence the in receptors eetr r aiyo iadgtdincanl hc epn otemjrihbtr neuro- inhibitory major the to respond which channels ion ligand-gated of family a are receptors .Bsiai a icvrdi eiia hmsr ffr trigfo h eut fa56,265 a of results the from starting effort chemistry medicinal a in discovered was Basmisanil ). A- A α - α Ashl rms sptniltetet o utpeidctos particularly indications, multiple for treatments potential as promise hold NAMs 5 ,Cgiin E,EEG PET, Cognition, 5, α uuis( subunits A - α eetr ihaslcieadsf compound. safe and selective a with receptors 5 α 1, α otiigrcposadi rciia tde aedemonstrated have studies preclinical in and receptors containing 5 α 2, A - α α, α α or 3 eetv Aswtotsft iblte e obasmisanil to led liabilities safety without NAMs selective 5 A two and - α Asmyb eeca o lhie’ ies and Disease Alzheimer’s for beneficial be may NAMs 5 α A β, )aduulythe usually and 5) β - 4 α n one and ciiyehne ucinlrcvr fe stroke after recovery functional enhances activity 5 uuis aycmonsi lncluea anxi- as use clinical in compounds Many subunits. α I Dwo ta. 06,MK06(tc tal., et (Atack MRK-016 2006), al., et (Dawson 5IA A eetrsbnt htasml spentamers, as assemble that subunits receptor γ uui Osn&Sehr,20) GABA 2009). Sieghart, & (Olsen subunit A γ eetrsbye odtriethe determine to subtypes receptor uui.Teecmonsare compounds These subunit. 2 A A - - α α eitdtncihbto in inhibition tonic mediated 5 uui-otiigreceptors subunit-containing 5 3 A ]flmznlcompetition- H]-flumazenil α - α I n R-1 were MRK-016 and 5IA Ashv lobeen also have NAMs 5 A A - Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. n=7 and 7) = (n temo h eunet etasrbd(e uprigTbeS) apdadpl()tie cRNA tailed poly(A) mMessage and the Capped using by protein S1). desired Table the Supporting up- encoding site (see plasmids promoter corresponding transcribed linearized polymerase the from be the of synthesized to keeping end were 3’ enzyme sequence transcripts the restriction the at appropriate of linearized the were stream with oocytes: human constructs cDNA laevis The subunit techniques. Xenopus cloned receptor standard into by microinjection USA) expressing vitrogene, for website, RNA GABA oocytes human of the different preparation laevis and on Cloning described Xenopus methods of the GABA Voltage-clamp following 2.1.3 France) l’Evescault, (Celle SA www.neurofinsdiscoveryservices.com. (10 Cerep basmisanil Eurofins of Binding screening Selectivity 2.1.2 10 of presence the [ in of measured section. nM was analysis binding 1 statistical Nonspecific with IC incubated basmisanil. were 4513. of Membranes concentrations ten experiments. and analysis. competition-binding GABA statistical 15-4513 for the basmisanil Ro in of described as affinity and The UK) Surrey, Guildford, (IDBS; [ ActivityBase of using inhibition percentage The GABA [ diazepam. human nM expressing 1 membranes of in inhibition measured The was 2009). misanil al., et (Ballard previously al., et (Malherbe described previously as Switzerland) GABA Rotkreuz, 2008). 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Data may be preliminary. utpsadtu ol eue o l uui obntost e fteoctswr omlyresponding. normally were oocytes the if see to combinations for subunit of all concentrations for effective used Maximally be could thus and subtypes nml.Tepretg frcpo cuac y a acltdfrom: calculated L-655,708- was basmisanil-treated of and (y) vehicle- hippocampus occupancy of the receptor hippocampus in of of the radioactivity percentage determination in earlier the radioactivity The the described (S), the for as binding animals. autoradiography, from used specific for subtracted was the was used plasma obtain was animals divided To Blood brain treated were the 2009). ice. 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(Purina ad kg; food 7-10 of fascicularis; regimen male), daily (Macaca test: g, avoidance macaques approach 450-500 cynomolgus (social test: rats Male avoidance Fischer approach F-344 male), Sprague-Dawley social animals g, female), male). The for 220-250 and water. g, rat maze: male and 170-180 water stimulus g, (Morris food female; rats 200-220 to g, Hooded access experiments: (20–23 Lister libitum 180 (PTZ temperature ad rats occupancy: allowed controlled Wistar (receptor were a were They rats experiments at cycle. the light/dark rooms in h Care holding 12 used Animal a separate Institutional with in Alto and guidelines. Palo housed 65%) NIH Roche the group the with Veterinary were by accordance Cantonal Rats approved Cantonal in the was were and by NHPs and approval Federal in Committee prior Swiss task Use received the retrieval and and with object regulations complied The AAALAC and (Basel) Office. research Roche animal Hoffmann-La on F. laws at performed Experiments Animals 2.2.1 Studies Vivo In 2.2 plate). per value (mean 53% and 42% between α 1 β 2 γ ,30n for nM 300 2, A - α lce -5,0 1 gkg mg (10 L-655,708 blocker 5 α -1 2 β A ru)ad4 i ae eevditaeosy01mik f[ of mCi/kg 0.1 intravenously received later min 45 and group) μ 3 - g/kg). γ α eetrocpnyi as [ rats, in occupancy receptor 5 or 2 α 3 β β- 3 γ C eeue o ahsbntcmiain .. 0 nM 100 i.e., combination, subunit each for used were CCM -1 n 1 and 2 ort n . Lkg mL 2.5 and rats to 3 ]R 541 n ru faias( )rcie the received 4) = (n animals of group one 15-4513 H]-Ro μ for M -1 7 ..3 i ro ordoiadijcin(e (Atack (see injection radioligand to prior min 30 i.p. ) α 5 β 3 3 ]R 541 a mlyda radioligand a as employed was 15-4513 H]-Ro γ .Teihbtr ffcsof effects inhibitory The 2. -1 oNP i rlgvg.Pre-treatment gavage. oral via NHPs to -1 ..( e oegroup, dose per 4 = (n p.o. ° ) uiiy(55- humidity C), 3 β- ]R 15-4513 H]-Ro C ranged CCM β- CCM Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. 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[ the Quanti- mL Radiochemistry with scan. ng 2.3.3 spectrometry, PET 1.0 mass dose at chromatography-tandem post quantification liquid the by of before performed limit min was lower 30 basmisanil starting plasma timepoints of different fication four at performed was sampling eeelm Oswr eetdt rvd ra oeaeo h ri,adecmasarneo levels of range a encompass and brain, the of ventral coverage and broad pons provide medulla, to midbrain, GABA selected putamen, of were hippocampus, cortex, accumbens, frontal ROIs amygdala, caudate, medial lobe, cortex, pallidus, cerebellum. temporal frontal MR globus dorsolateral lobe, subject’s cingulate, cortex, parietal the anterior orbitofrontal lobe, cortex, to brain occipital insular warped co- gyrus, template considered: nonlinearly parahippocampal (ROIs). and MNI152 were was interest ROIs The 2011) segmentation of following regions al., The 2006). matter of et al., definition (Tziortzi grey automated et atlas enable (Grabner extraction, (CIC to (MNI152, atlas image brain space associated underwent reference and standard image image a and MRI to extraction structural registration brain (version for subject’s FSL functions of Each 2004) use al., segmentation makes et image Smith and for registration. http://www.fil.ion.ucl.ac.uk/spm) 2005; USA), Neuroimaging, implemented and al., MA, for is et Centre Natick, MIAKAT Trust (Jenkinson (Wellcome Inc., SPM5 3.3.1). 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All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. 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All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. 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No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. 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No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. fPZ h w ihrdsspoue dnia ffcs(al )adhdsmlrpam concentrations plasma 2). (Table similar expected had as and convulsions, dose 2) tonic sub-threshold (Table showing a effects rats of identical of administration kg number produced following mg doses convulsions 100 higher tonic to two exhibited rats rats The female eight PTZ. In of of above. out (Table described kg five test experiments mg 5) for binding PTZ 100 Figure vivo 60-70% the of in approximately in the dose are in activity a concentrations observed proconvulsant at plasma maximum exert these the are at not which estimated did ceptors, occupancies 5) receptor Figure The concentration; 2). plasma kg free mg 30 nM test and kg 2452 PTZ concentration) mg the plasma 100 occupancy. in to free receptor concentrations up nM relevant estimated basmisanil therapeutically 50% of at concentrations and doses basmisanil 30 plasma Single of about free activity to Active proconvulsant corresponded of which S6). Lack Table 5), 3.6 (Figure (Supporting nM dose 345 with and increased 156 object were exposure kg the mg plasma 30 kg of and total trials 1 mg The the difficult 10 with adult paradigm during and this reaches in 3 in response first the correct task at of task retrieval percentage retrieval the object improved significantly the Basmisanil in correct macaques percent 65%. cynomolgus improved and GABA 45 Basmisanil of between occupancy be 3.5 to 100% mL estimated to ng was corresponds experiment 13 maze actually +- water curve kg the 903 the mg during 10 of of time-dependent occupancy at level maximum and basmisanil dose- asymptotic maximal of were administration the a the groups reached after dose min parallel and 30 in S5) determined Table quadrant as (Supporting platform basmisanil previous 4A). of the kg (Figure concentrations in quadrants mg Plasma spent opposite 10 time and the percent at right during in Basmisanil the left, increase quadrant in to an each groups quadrant. compared by in diazepam-treated revealed platform and time as of the vehicle deficit of learning between in diazepam-induced quadrant amount difference disrupted spent equivalent platform significant significantly time a an the was Diazepam spent percent in there 4A). spent Moreover, subjects (Figure since time trial. the quadrants probe percent position of opposite in platform learning and new spatial increase right demonstrated significant the (Supporting group left, the vehicle speed by to The swim indicated compared trial. on as seventh the effect position, before platform no removed had was a platform and reduce induced The significantly pathlength and not did and position Basmisanil latency platform S2). S2). in the Figure Figure (Supporting find increase speed to diazepam-induced swim pathlength the in increase and significant in latency but test the small maze increased water significantly Morris Diazepam the in mL impairment ng learning 125 +- spatial 544 rats diazepam-induced of attenuated level (EC Basmisanil plasma occupancy and 3.4 receptor 5% half-maximal +- equation plasma producing (i.e., 77 the equation for of against Hill concentration) occupancy the occupancy by maximal plasma receptor data a specific the in Fitting plotting 3B). resulted by (Figure 1) analysed individual further each of were basmisanil data of kg Binding concentration mg 6%. (100 +- dose 70 highest by The kg 3A). (Figure mg manner (3-100 dependent basmisanil with Pre-treatment -1 Spotn alsS n 8.Tepstv oto,F74,sgicnl nrae the increased significantly FG7142, control, positive The S8). and S7 Tables (Supporting -1 ttlpam ocnrto f110n mL ng 12100 of concentration plasma (total -1 oe Fgr B.Bsiai xiie nivre -hpddose U-shaped inverted an exhibited Basmisanil 4B). (Figure doses -1 nml as(oa lsacnetaino 84n mL ng 3864 of concentration plasma (total rats male in -1 -1 nfml as ttlpam ocnrto f54 gmL ng 5840 of concentration plasma (total rats, female in .. erae h idn f[ of binding the decreased p.o.) -1 oe rdcn omre mrvmn nperformance. on improvement marked no producing doses 15 -1 Fgr ) sn iue3 n suigthat assuming and 3B Figure Using 5). (Figure -1 eue pcfi idn ntehippocampus the in binding specific reduced ) -1 09n repam concentration; plasma free nM 5079 ; -1 ..sgicnl teutdthe attenuated significantly p.o. 50 -1 ). 39+ Mfe plasma) free nM 5 +- (379 3 ]R 541 nadose- a in 15-4513 H]-Ro -1 28+ 2n free nM 52 +- (228 A - α receptors, 5 -1 1622 ; α re- 5 -1 ; Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. umdPTiae n isetm ciiycre a eue eaiet aeiesas indicating scans, baseline to both relative in reduced heterogeneity was basmisanil, curves of activity administration time [ Following [ tissue 6A). of and cortical Figure blocking and images and amygdala 2017); accumbens, PET PET nucleus al., summed hippocampus, using et the volunteers in (Myers signal healthy regions highest in the with determined receptors, containing was basmisanil by occupancy receptor [ of degree The ολυντεερς ηεαλτηψ οφ GABA βραιν non-selective the and ΓΑΒΑ compartment Δοσε-δεπενδεντ 2006), social 3.8 the Prinssen, 3). in (Table estimated & time area with the (Nicolas hidden S9) decreased the expected significantly Table in 19-4603, As (Supporting time Ro found the control, positive were increased 77%. a 5) to as Figure used 50 concentrations; NAM of plasma in spent occupancies free time kg nM receptor the mg increase 100 3059 or to compartment to up social dose (174 the any in at spent area time hidden the the reduce avoidance significantly approach not did social Basmisanil the in effect rats anxiogenic in an have test not did Basmisanil 3.7 GABA [ published previously with utemr,tesailtpgah ftebt ogmabn ffcssgicnl iee o basmisanil for differed significantly was effects effect band band gamma gamma to effect). to beta low-frequency the beta for of the difference significant topography of (no spatial frequency midazolam the peak to Furthermore, the compared However, basmisanil for 5E). higher Figure significantly Supporting band; gray 7A, Hz; 8.7 – (6.2 range frequency alpha low to 19.1 theta- 7.3 the by in 7A) 6B). power Figure Figure relative in increased depicted significantly are Basmisanil assessments EEG during ranges occupancy twice mg and 4030 (240 h basmisanil with 4 GABA Treatment mida- of of and basmisanil. occupancy dose with high acute treatment to an sub-chronic led after of daily) days baseline, 14 at after assessments and EEG zolam, state volunteers resting underwent healthy volunteers healthy in Twelve basmisanil receptor of in signature delay Electrophysiological significant 6B). 3.9 (Figure no concentration i.e., plasma EC relationship free be an nM direct 67 can with a to concentrations concentration, suggesting corresponding plasma plasma concentration model, and to occupancy Emax compared receptor simple occupancy between a relationship by The described S11). Table (Supporting yadces frltv oe tlwfeunisi h ihteat o lh ad (8.2 bands alpha low to theta high the in characterized frequencies midazolam, low of at that power to relative opposite of qualitatively -14.4 was decrease basmisanil a and by open by induced eyes signature 5C). pattern as spectral Figure well spectral the (Supporting as that The conditions assessments suggested four separately dose multiple-comparison all conditions post four across survived h for the similar 4 pattern of peak of and is spectral each low-frequency frequency h Analysing similar 1 the conditions. a peak combined closed only confirmed analyses eyes Hz; we above while First, the (13.5–38.0 5D), Second, effects. range Figure correction. these (Supporting frequency of nature power gamma the absolute low investigate further to to beta- analyses the in 34.9 power relative decreased 11 11 ]R 541 idn nalrgoswsosre fe diitaino amsnl h calculated The basmisanil. of administration after observed was regions all in binding 15-4513 C]-Ro consistent signal structured spatially a displayed baseline at brain the of images PET 15-4513. C]-Ro ± ± ± . z hc a eo h enapapa rqec f93H.Frhroe amsnlsignificantly basmisanil Furthermore, Hz. 9.3 of frequency peak alpha mean the below was which Hz, 0.4 A .3H)b 18.9 by Hz) 3.13 .3)adicessa ihrfeunisi h eabn (18.4 band beta the in frequencies higher at increases and 3.73%) - α eetrocpnya h ihs oeo 00m amsnlrahd9%i h hippocampus the in 94% reached basmisanil mg 1000 of dose highest the at occupancy receptor 5 11 ± ]R 541 GABA 15-4513 C]-Ro 4. gmL ng 842.3 ± ± .%cmae opetetetbsln.Tepa rqec fteeetwas effect the of frequency peak The baseline. pre-treatment to compared 3.3% .4 oprdt h r-ramn aeie epromdasre fadditional of series a performed We baseline. pre-treatment the to compared 4.24% 11 ]R 541 E aaadtekondsrbto fGABA of distribution known the and data PET 15-4513 C]-Ro -1 eetrocpnyatr1h92 h 1 after occupancy receptor ; A A Α - - α α εετροςπνψβ αμσνλι ηυμαν ιν βασμισανιλ βψ οςςυπανςψ ρεςεπτορ -α5 α eetrbnigb amsnl oedpnetdces in decrease dose-dependent A basmisanil. by binding receptor 5 eetr of receptors 5 -1 Tbe3.Pam ocnrtosbten44t 28n mL ng 7288 to 414 between concentrations Plasma 3). (Table 16 > 0 pam xouea 3284 h 1 at exposure (plasma 90% 50 fapoiaey51n mL ng 541 approximately of ± %adatr4h94 h 4 after and 3% ± .6H,32.2 Hz, 6.06 ± ± ± -1 .% exposure 1.5%; .4)(Figure 7.04%) 8. gmL ng 887.9 A oa plasma total - α ± subunit- 5 .2Hz, 4.12 -1 -1 A Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. Blade l,20) nNP amsnlipoe efrac nteojc erea aka estimated at task retrieval object the kg GABA in of performance mg activation improved 10 reduced basmisanil via NHPs at mediated In basmisanil be GABA Reversal 2009). to of al., impairment. proposed administration et diazepam-induced been (Ballard acute has the impairment attenuated test, diazepam significantly memory of 45-65%) and between occupancy learning receptor spatial dose-dependent the demonstrated In basmisanil 40- rats. Overall, in about 77%. vivo was is in which compound engagement RO4938581, this target RO4938581 with of achieved of selectivity occupancy occupancy binding receptor versus receptor lower selective higher the 129-fold apparent to and The due previous 2009) be versus our al., may fold with et basmisanil experiments (Ballard to similar [ compound compared In the Information: 3). by Supporting (Figure occupied (see curve 6 binding equation in-vivo and α basmisanil values the these with From agreement [ 1999). of al., 17-33% et 4513), Sur and 1991; radiolabel the al., of et selectivity limited the hippocampus, by explained be can the or of This of abundance 23% basmisanil. relative Thus, by the 77%. occupied at be maximum not a could approaching basmisanil using for curve experiments binding binding in-vivo GABA Our at far. efficacy so and described affinity development clinical in preference selective α GABA-induced highly inhibited it the GABA since bining selectivity, expressing functional cells showed in also current Basmisanil GABA assays. human recombinant binding of competition site versus binding selectivity 90-fold BZD than the to bound Basmisanil Basmisanil of Profile Preclinical Conclusions and Discussion any in events, There adverse studies. to EEG due and discontinuations PET the temporary in or evaluated studies discontinuations subjects events, cohort. EEG healthy adverse the severe and in no PET tolerated were well the and safe in was Volunteers Basmisanil Human Healthy in (within/between 5F). Safety Figure receptors 3.10 Supporting targeted 7B, of Figure pattern 0.128/0.003; expression = in r difference correlation the cross-subject drug reflecting likely midazolam and h Hswr elh,uipie dlswihpromda oe cuaydet akdffiut.In difficulty. task contrast to In due accuracy shown). lower GABA not at decreasing (data performed strongly which performance condition, adults cognitive healthy unimpaired GABA of had healthy, this rodents enhancement so were and non-impaired shown explanation model NHPs In not not pharmacologically-impaired possible the was a have A deficit. doses in RO4938581, higher cognitive assessed dose-dependently. of was existing and basmisanil increased effect an study of concentrations reverse rodent lack plasma the to The since in study. that rodent exposure, be the of may in lack observed to not was due which U-curve inverted an mareti asadi neeuiefnto aki NHPs. in task function executive an learning in spatial and pharmacologically-induced rats in in properties impairment enhancing cognition exhibited basmisanil summary, -eetv A,R4351 pt 0 ftebnigstsof sites binding the of 90% to up RO4938581, NAM, 5-selective GABA potent and selective most the represents basmisanil receptors, subunit-containing 3 α 2/ α A ag ewe 1ad1 Csl ta. 01 aiga ta. 93 ur ta. 96.I the In 1996). al., et Quirk 1993; al., et Hadingham 2001; al., et (Casula 15 and 11 between range 5 - α eetrocpnybten3-0,wihi iia oterdn td.Hwvr hr was there However, study. rodent the to similar is which 30-50%, between occupancy receptor 5 α -and 1- α -otiigrcposwr on oacutfr1-6 falGABA all of 16-26% for account to found were receptors 5-containing 3 ]R 541 sepce obn to bind to expected is 15-4513 H]-Ro α -otiigrcpos(alr ta. 09.I h rsn td,bsiai a 68- was basmisanil study, present the In 2009). al., et (Ballard receptors 2-containing α -and 1- α α -and 2- 1, α A ,and 2, α - α -otiigrcpos o o1-53pbihdant ratios affinity published 15-4513 Ro For receptors. 2-containing e a iteo oeeta h te eetrsbye.B com- By subtypes. receptor other the at effect no or little had yet 5 α -otiigrcposrsetvl,wihsget h h maximal the why suggests which respectively, receptors 1-containing 3 α -o1-53a ae o GABA for label a as 15-4513 H-Ro uui-otiigrcpos sdmntae yorflumazenil our by demonstrated as receptors, subunit-containing 3 A 17 - α ciiymyntb eeca o onto.In cognition. for beneficial be not may activity 5 α -and 1- α 3 -o1-53i h ipcmu were hippocampus the in 15-4513 H-Ro -otiigrcpos hc si good in is which receptors, 2-containing A A A - - - α α α As nldn basmisanil including NAMs, 5 essGABA versus 5 ih5n ffiiyadmore and affinity nM 5 with 5 A 3 - -o1-53bnigsites binding 15-4513 H-Ro α eetr eutdi a in resulted receptors 5 A eetr (McKernan receptors A A A -1 - - α α α receptors 5 3 1, (estimated A in NAM 5 ]R 15- H]-Ro α ,and 2, α 1/ α 5 Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. tde hudivsiaehwmdlto ftedffrn Z-estv eetrsbye,b subtype by subtypes, receptor BZD-sensitive frequency different GABA of on the terms work in of aligned prior modulation well with are how change basmisanil investigate of the effects should of EEG directionality 2000), studies the al., Thus, and et detail. affected Whittington 1999; in bands al., understood et syndrome, not (Traub et tissue are (Dup15q cortical but CNVs (Visser in involving loops GABA rodents conditions feedback excitatory-inhibitory in genetic in recurrent rare SNPs activity GABA for and beta-band reported. and activity been 2018) reduce GABA al., beta-band including to GABA et syndrome) between Non-selective Smit shown Angelman links 2002; 2003). been al., al., genetic have et et Furthermore, (Porjesz Visser 19-4603) activity 1996; of Ro al., 2003). decrease et DMCM, a al., Malizia and 1992; marker, (e.g. al., pharmacodynamic et NAMs studies. established (Friedman clinical an frequencies future is lower for which at activity readout in beta-band pharmacodynamic decrease in a basmisanil a increase as that and EEG demonstrate range) suggest results GABA and frequency These Non-selective function alpha range). brain to frequency on (theta gamma effect power to an EEG (beta has frequency power lower GABA EEG of in target frequency modulation increase specific higher negative an a that cover to revealed adequately volunteers, leads healthy to tors in as basmisanil way of a assessment such EEG in selected be allow to to will trials compared relationship clinical concentration-occupancy occupancy. occupancy drug future the receptor for Understanding in regimens concentration. dosing delay plasma significant free nM simple no 67 a to i.e., by EC relationship, described an with be direct concentration, can a plasma concentration brain suggesting plasma the model basmisanil enters and Emax basmisanil occupancy that dogs receptor indicates between in volunteers, GABA relationship human the studies of healthy toxicology binding in GLP the vivo blocks of in in and symptoms signals obtained convulsion data, reveal and in imaging mL not signs PET shown ng clinical selectivity did 4590 of to receptor readings needed lack up high the EEG those exposures the with exceeded six-fold. at mirroring line study approximately in abnormalities, DDI by and epileptiform vitro and mean trough as MAD such a at the effects, in occupancy pro-convulsive in resulting receptor observed 14500 daily full values to twice concentration for raised substrate plasma administered 8520 CYP3A maximum were and sensitive The (26.5%) mg a 6040 as In 1000 (31.3%), basmisanil 4110 and Information). of variation), of Clinical 370 mL coefficient Supporting 160, ng (27% (see (32.2%) 2120 80, basmisanil of of concentration of plasma exposures doses included maximum was plasma study, monitoring high EEG MAD covering exposures, the studies therapeutic at I humans Phase in risk in pro-convulsive of absence Volunteers confirm the To Healthy of in risk pro-convulsive Basmisanil either the of GABA over-estimate Profile antagonize and Clinical that conclusions drugs suggested 2009). positive of (Loscher, been false activity has question to proconvulsant It in the lead molecule system. toxicology assess may GABAergic GLP to indirectly the the test or and in PTZ test directly PTZ and PTZ the through rats the of activity in between application synergistic weeks discrepancy that to 26 The related to S4). up be Figure may for Supporting studies exposures (see supra- plasma dogs at following supra-therapeutic in alterations only weeks at EEG 39 basmisanil no activity and other of proconvulsant convulsions at administration no exhibited shown binding oral have basmisanil daily to studies due pharmacology test, although safety be PTZ and S10); may toxicology Table the which GLP and exposure, In S3 plasma was Figure high tested. gating (Supporting therapeutic 40-50% not sensorimotor of Moreover, of were importance occupancy 5). doses receptor critical of (Figure higher at efficacy the occupancy basmisanil functional by underlining receptor and affected potential, affinity estimated not binding proconvulsant with in and cognition selectivity anxiogenic improve subtype lacked to basmisanil shown concentrations 30-65%, plasma at Importantly, -1 epciey(i elh outesprds) na taoaoeDIsuy lsalevels plasma study, DDI itraconazole an In dose). per volunteers healthy (six respectively , A A As ..BD,aekont nueacaatrsi E intr ihan with signature EEG characteristic a induce to known are BZDs, i.e. PAMs, eetrrltdcagsi eabn ciiyaetogtt eetmdlto of modulation reflect to thought are activity beta-band in changes receptor-related -1 . 50 A fapoiaey51n mL ng 541 approximately of - α rfriglgn [ ligand preferring 5 A eetrgns(rhihe l,21a rhihe l,21b have 2019b) al., et Frohlich 2019a; al., et (Frohlich genes receptor 18 11 ]R 541 nads eedn anr The manner. dependent dose a in 15-4513 C]-Ro ± -1 80n mL ng 1810 oa lsacnetain corresponding concentration, plasma total α utps ncnrs,preclinical contrast, In subtypes. A -1 eetrmdltr.Future modulators. receptor n=1 elh volunteers). healthy 12 = (n A eetrgenes receptor A - α recep- 5 A Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. hnZ,CagC,Li A leeR le W(05.GABA (2005). RW Olsen & R, Olcese TA, GABA Leil CS, Chang ZW, Chen GABA K the Maubach at K, ligand the Martin site for 304 PB, responsible Science Wingrove residues networks. GV, acid cortical Pillai amino FA, in GABA Bromidge oscillations MA, Neuronal a Casula (2004). RO4938581, A Draguhn 85 & (2013). Pharmacol G, JP Biochem Buzsaki rats. Redrobe in & properties autoinduction L, CYP1A2 F Badolo 25 Chaumont England) C, (Oxford, de Bundgaard psychopharmacology L, of Dauphinot Journal PL, mice. receptor syndrome Pereira GABA-A PsychopharmacologyDown A, the mGlu5 Duchon comparison. of the B, a targeting of Delatour effect cognition: The J, and (2005). Braudeau anxiety W of Spooren tests & R, rodent 179 Porter in (Berl) J, MPEP Huwyler antagonist E, J Prinssen receptor Basile ML, Woolley JA, TM, Vivian Ballard E, GABA Borroni at E, acting Prinssen enhancer cognitive F, Knoflach TM, Ballard DC 331 Evans Ther AD, Exp 3-tert-butyl-7-(5-methylisoxazol-3-yl)-2-(1-methyl-1H-1,2,4-triazol-5-ylmethoxy)-macol Rodrigues GABA of a D, (MRK-016), properties O’Connor pyrazolo[1,5-d]-[1,2,4]triazine vivo en- KA, for L-655,708 Wafford in selective KA, and (2006). dose Maubach GR a JR, Dawson at Atack & proconvulsant RM, not is McKernan but KA, 51 Wafford rats Neuropharmacology GR, in cognition Seabrook hances PJ, Bayley JR, Atack GABA the of pharmacology clinical and agonist Preclinical (2010). JR Atack References first GABA our for biomarker and pharmacodynamic are PET useful studies using a Further be engagement humans. would in target this basmisanil activity. of direct whether engagement determine demonstrated target to functional studies required for clinical evidence GABA provided efficacy Our estimated into study at EEG translates profile basmisanil 30-65%. safety of adequate around profile an of pharmacological with vitro together in paradigms cognition the in that showed studies preclinical Our when power Conclusions and absolute and acute the Summary for identify to findings, significant generalize these also basmisanil. not confirm findings dose with was to the treatment high need range if long-term difficult a studies understand frequency Future it Only and beta makes frequencies. relationship 2013). across pharmacokinetic/pharmacodynamic the which testing Siegel, in used, multiple & effect was for (Hipp correcting the electrodes activity Furthermore, 19 band with there gamma tested. (n=12), EEG study low clinical was to a subjects artifacts ( of only high-frequency number and The clean condition, considered. to activity. be placebo should oscillatory no which large-scale limitations, was the has in study changes EEG similar The qualitatively to lead can ligands, selective > 0 eetrocpny nsbauetetetrgm floig1 aso amsnldsn)was dosing) basmisanil of days 14 (following regime treatment sub-acute in occupancy) receptor 90% A α I.Pamclg hrpuis125 therapeutics & Pharmacology 5IA. eetrcl-ufc ubri eou avsocts oeua hraooy68 pharmacology Molecular oocytes. laevis Xenopus in number cell-surface receptor : 218-229. : 470-484. : A 1023-1029. eetr erce 77 Neurochem J receptor. A α utp yaslcieivreaoitrsoe ontv ect in deficits cognitive restores agonist inverse selective a by subtype 5 α α uui-otiigrcpos scohraooy202 Psychopharmacology receptors. subunit-containing 5 uui idn eetvt fL6578 ezdaeiebinding benzodiazepine a L-655,708, of selectivity binding subunit 5 : 11-26. A 19 : receptor 445-451. α utp-eetv nes gns.JPhar- J agonist. inverse subtype-selective 5 : 1363-1369. A A tal. et , eetrascae rti regulates protein receptor-associated receptor α A -otiigGABA 5-containing tal. et , α : ouao,dsly strong displays modulator, 5 1030-1042. 20) O988,anovel a RO4938581, (2009). α utp-eetv inverse subtype-selective 5 tal. et , tal. et , A 20) dnicto of Identification (2001). - α eetroccupancy receptor 5 : 1926-1929. 20) nvitro In (2009). 21) Specific (2011). : A 152-159. A - α : receptor 5 receptors. 207-223. Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. ect npelnclmdl.JPamclEpTe 366 Ther Exp Pharmacol J models. surfaces. A preclinical Lewis scalp in M, and deficits Higashino skull K, GABA brain, Hazama novel of N, estimation a Nakanishi muscle MR-based M, ocular Nakanishi BET2: and S, S Kawaharada cranial Smith & from M, activity Pechaud gamma-band M, neuronal 7 Jenkinson Neurosci Dissociating Hum Front (2013). EEG. M in Psy- Siegel activity & tomography. JF, emission Hipp positron and autoradiography 108 Preparation, (Berl) chopharmacology binding. receptor benzodiazepine recombinant cDNA of of Cloning acid pharmacology (1993). PJ benzodiazepine Whiting & the CI, Ragan human ligand-receptor KJ, 6 Palmer encoding of Neuroimage B, sequences Bourdelles imaging model. Le Parametric P, region Wingrove KL, reference (1997). Hadingham simplified VJ a Cunningham and using & atlasing PET SP, Symmetric in Hume Comput binding AA, Comput (2006). Image Lammertsma DL Med RN, Collins adults. Gunn older & in J, hippocampus Pruessner the 9 to D, Interv application Assist Smith an MM, segmentation: Budge based C 10 model AL, Autism Hyde Janke Mol S, G, syndrome. Huberty Grabner Dup15q C, in DiStefano biomarker EEG V, MC the Saravanapandian Hoener 85 underlying LT, Psychiatry H, Reiter Biol Purtell J, Genotypes. Frohlich P, Between Garces Differs GABA Syndrome LM, Angelman Bird (1997). in MT, U Phenotype Miller Rudolph J, & Frohlich H, Clin formation Mohler receptor DK, time. for and Johnson prerequisite concentration, essential D, plasma Benke dose, JM, Fritschy of JS effect Harmatz MD, diazepam: 52 Charlton oral Ther CM, of Pharmacol Metzler pharmacodynamics of GR, and Ac- Modulators Peters Antidepressant cokinetics Negative DJ, Rapid by Greenblatt Stress H, (2015). Friedman Chronic SM After Thompson Strength & Synaptic TA, GABA Excitatory LeGates Containing of MD, Restoration Kvarta Hoffmann-La and AM, F. tion inventors, Dyke Van Patent (2013). J, International P Fischell 25. Waldmeier acids. April isoxazolyl-methoxy-nicotinic & 2013 of A, preparation A1. Thomas the 2013/05712 a for H, WO 7142, Process Iding FG assignee. S, by AG, Hildbrand induced Roche SP, anxiety Hanlon Severe P, (1983). 2 Dott M Lancet Amin receptors. & benzodiazepine G, for Paschelke ligand R, Horowski R, AM Dorow MacLeod BJ, Everitt M, 316 Cobain reviewers. for N, peer selective Collinson and agonist KA, authors for Maubach guidance GR, simplified Dawson MA and GABA- Giembycz updated excessive 175 CH, II: Pharmacol Reducing George reporting J JR, their Br Docherty and (2010). G, analysis ST and Cirino Carmichael 468 design S, Nature & Alexander stroke. MJ, I, after Curtis recovery Mody functional SE, promotes Macisaac inhibition tonic BS, mediated Huang AN, Clarkson ali ,FreL itnJ,Hl ,&SdalG(92.[ (1992). G Sedvall & H, Hall JE, Litton L, Farde C, Halldin : A 1335-1345. eetr.Mlclrpamclg 43 pharmacology Molecular receptors. : A 58-66. α eaiealsei ouao nacsln-emptnito n mrvscognitive improves and potentiation long-term enhances modulator allosteric negative 5 A : 139-150. : eetr.Nuoscohraooy40 Neuropsychopharmacology Receptors. 987-993. α uui-otiigGABA subunit-containing 5 α : and 2 16-22. α am-mnbtrcaiArcpo uuisadcaatrzto of characterization and subunits receptor acidA gamma-aminobutyric 3 : 338. nvivo in : 970-975. : 98-99. A α ersine81 Neuroscience . eetr nacscgiin hrao x Ther Exp Pharmacol J cognition. enhances receptors 1-, 20 α 2-, : 58-65. α - and 3-, : 11 2499-2509. ]o1-53 iadfrvsaiainof visualization for ligand a 15-4513, C]Ro tal. et , : : α 29. -otiighuman 5-containing 1043-1053. : 279-287. tal. et , tal. et , 21a.Electrophysiological (2019a). tal. et , : tal. et , A 305-309. -receptor tal. et , : 752-759. 21) ONO-8590580, (2018). 21b.Mechanisms (2019b). 21) Experimental (2018). 20) ninverse An (2006). 19) Pharma- (1992). α- γ- uui san is subunit aminobutyric β- carboline α 5- Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. ihl E omsA 20) oprmti emtto et o ucinlnuomgn:aprimer a neuroimaging: functional for 15 tests Mapp permutation Brain Nonparametric Hum (2002). 37 examples. AP Metab with [ Holmes Flow & of Blood TE, Quantification Cereb Nichols J (2017). humans. RN in Journal Gunn selectivity rat. regional & the and RA, in Comley learning spatial JF, studying Myers for procedure 11 water-maze methods a neuroscience of of Developments (1984). 61 Biometrika R potential Morris review. for jackknife-a implications The its and (1974). urine RG human Miller in metabolite 60 hydroxylated AC Pharmazie soluble Huber toxicity. BH, poorly renal Schaefer a EJ, of Woolf solubility navi- GE, the Pearce place MJ, impairs Rose benzodiazepine, SA, 24 Merschman anxiolytic research brain an Behavioural Chlordiazepoxide, rats. (1987). in RG gation Morris & N, McNaughton CI 7 with Ragan NP, Neuron brain Gillard PA, rat Cox from R, immunopurified Prince K, A Quirk RM, Bertoldo McKernan G, [ 152 Rizzo Neuroimage of M, subunits. measures Veronese alpha5 binding L, containing quantitative Rosso of DA, Barros reproducibility Riano CJ, McGinnity 2 down Disord of GABA model (2003). mouse K V Maubach a Vidal in 33 S, deficits Neurosci Garcia neuromorphological J R, and syndrome. functional Vidal rescues N, drug 35 inhibition Rueda Neuropharmacology receptor-mediated P, of VJ vivo. Martinez measurement Cunningham in C, direct brain PM, Martinez-Cue human Bloomfield the man: on SH, in effects Waters and quantification SJ, occupancy pharmacokinetic/pharmacodynamic Wilson site RN, zodiazepine Gunn neurokinin tachykinin AL, 73 human Malizia the pharmacology in Molecular pockets domains. binding transmembrane identical receptor JG not Wettstein 3 but MT, overlapping Zenner within C, interact Kratzeisen A, osanetant Marcuz C, Bissantz P, Malherbe J Hojo S, Obayashi predicting 200-208. T, for Okauchi relevance K, GABA its Kawabe of T, and Suhara activity 610 J, ligand Pharmacol drug Maeda receptor J proconvulsant Eur benzodiazepine of humans. the assessment in of Preclinical events 83 effects adverse Pharmacol chronic (2009). J and W Br Acute Loscher kindling. and (1984). properties SC proconvulsant Taylor & 7142: FG DJ, GABA Nutt native HJ, of Little Evaluation (2001). 413 HC Pharmacol Rosenberg J & Neuroim- Eur A, studies. Szabo receptor M, PET for Li model 35 tissue Metab reference Flow Simplified 4 Blood (1996). age SP Cereb Hume J & AA, function. Lammertsma sensorimotor and M lesion Brown ischemic M, Ganguly on 1601-1609. R, inhibition Janik tonic L, of Thomason reduction J, Chaudhuri EM, Lake : 153-158. : : A 667-676. 233-239. bnoizpn eetrb [ by receptor /benzodiazepine : 63-72. : 3953-3966. A : : eetrsbyeslciecgiinehnes urDu agt N Neurol CNS Targets Drug Curr enhancers. cognition selective subtype receptor 47-60. 359-363. : α 1-25. uui-pcfi nioishv nqepamclgclproperties. pharmacological unique have antibodies subunit-specific 11 ]R1-53uigpsto msintmgah.Snpe47 Synapse tomography. emission positron using Ro15-4513 C] : : 270-282. 39-46. : 1-11. 21 11 ]o541,aPTlgn o GABA for ligand PET a C]Ro15-4513, : 1-15. tal. et , : 11 : tal. et , 1736-1750. ]o541 GABA C]Ro15-4513 2137-2148. : tal. et , A tal. et , 951-958. eetr otiigan containing receptors 20) iulzto of Visualization (2003). tal. et , 19) GABA (1991). : tal. et , 1483-1491. 21) euigGABA Reducing (2013). 21) h ffcso delayed of effects The (2015). tal. et , 20) hrceiainof Characterization (2005). 20) etleatand Me-talnetant (2008). tal. et , A 21) Test-retest (2017). A α pcfi binding specific 5 eetrsubtypes receptor 19) Ben- (1996). A α α subunit. 5 subunit 5 receptors A α 5 : : Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. rmImdaeRlaeFruain faPol oul opud amsnl hog Absorption Through Basmisanil, Compound, 19 J Soluble Release AAPS Drug Poorly Testing. Characterising Dissolution a (2017). A and of Szepes Modelling & Formulations D, Bentley Immediate-Release P, Chalus from M, H Lindenberg Suppl NJ, 23 Johansen-Berg Parrott Neuroimage C, FSL. TE, Stillhart as Behrens implementation and CF, analysis image Beckmann MR 1 MW, structural and Woolrich 39 functional Mapp M, in Advances Brain Jenkinson Hum SM, activity. brain Smith SM oscillatory Malone to YYW, genes Ho liability psychiatric NG, 4183-4195. multiple Martin links JL, analysis Meyers ciation MJ, GABA Wright of DJA, modulation Smit and function, Structure, (2012). Rev ME GABA Pharmacol 287 Steinmann CVI: & Humans. Pharmacology. E, to Clinical Translation Sigel and in Basic Issues of Key Union Ligands: 70 International Function-selective excitatory-inhibitory and (2018). of Subtype- MM dependence Receptor Savic Age & (2012). W, OW Sieghart 33 Witte Aging & Neurobiol C, Redecker stroke. C, following 1208 Frahm balance Res C, Brain Bruehl rats. S, in Schmidt learning avoidance GABA active, at not selectivity but functional S passive, agonist Savic J, inverse Samardzic moderate I, Gavrilovic with R, and Furtmuller rolipram T, (Berl) Clayton inhibitors MM, Psychopharmacology PDE Savic Selective macaques. cynomolgus (2008). adult JA in Vivian & performance 196 retrieval A, Blokland object J, improve Prickaerts GABA sildenafil JL, of Basile potential K, 10 therapeutic novel Rutten Discov Drug benzodiazepines: Rev classical Nat Beyond subtypes. (2011). F receptor Knoflach & 221 U, (Berl) Rudolph Psychopharmacology rats. GP in Smith deficits IE, cognitive Jong (PCP)-induced de C, GABA Bundgaard of K, tion Frederiksen L, Elster JP, Redrobe D Mellilo F, GABA Tang of P, 35 site Leeson benzodiazepine S, the for Fletcher selective an P, Pharmacol of Behav Blurton Effects K, task. Quirk acquisition (2014). repeated MN incremental Reed an & in deficits DE, 25 Tosto learning MK-801-induced HC, on Hunsberger agonist CC, Rudy JM, GABA Povroznik a and EEG T human Foroud the 99 DB, of A Chorlian frequency K, 101 GABA Wang beta Neuroscience HJ, the (2000). Edenberg between brain. L, G rat Almasy adult Sperk B, the & Porjesz in W, subunits Sieghart 13 A, of distribution Wieselthaler chemical C, Schwarzer S, rats: Pirker of strain GABA high-anxiety 56 (2009). a Neuropharmacology W of cology. Sieghart behavior 184 & approach-avoidance RW, (Berl) Psychopharmacology Olsen Social ligands. receptor (2006). benzodiazepine EP of Prinssen effects & LB, Nicolas : : : : S208-219. : : 836-878. 1331-1335. 331-335. : 40224-40231. 643-648. 3729-3733. A α eetr yR4351atnae iceesbcrncaderypsntlphencyclidine postnatal early and sub-chronic discrete attenuates RO4938581 by receptors 5 : 141-148. A : eetr:Sbye rvd iest ffnto n pharma- and function of diversity provide Subtypes receptors: : A 685-697. 1356-1363. : eetr hc oti the contain which receptors 827-836. 22 A tal. et , A eetrgn ou.Po alAa c S U Sci Acad Natl Proc locus. gene receptor eetr containing receptors : 150-159. tal. et , tal. et , 19) [ (1996). tal. et , : 451-468. tal. et , : α uui.Neuropharmacology subunit. 5 65-74. 20) W-2,acompound a PWZ-029, (2008). 20) ikg disequilibrium Linkage (2002). : 815-850. 3 ]-5,0,anvlligand novel a H]L-655,708, 21) eoewd asso- Genome-wide (2018). A A 21) eaiemodula- Negative (2012). eetr:immunocyto- receptors: eetr.JBo Chem Biol J receptors. α uuis improves subunits, 5 α tal. et , 5GABA A (2004). inverse A A : Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. GABA ( fects. ( (symbols) mean shown. as not shown are are size graphs the in Data 2 Figure 1,1-Dioxo-1,6-thiomorpholin-4-yl)- pyridin-3-yl 445.469; weight 1. Figure Legends Figure A Penna EM, Bridgwater SC, 4. eNeuro Haffey Mice. DS, in Wang Ketamine J, α Antagonist Yu Receptor NMDA AA, the Zurek of rhythms: Effects Side Inhibition-based the TD without Gould Action (2000). P, Georgiou EH SR, for Krimmel 38 Buhl JN, Modulator Psychophysiol Highland & J ME, Int Nelson B, P, dynamics. Zanos Ermentrout network on N, observations Kopell mathematical effects and RD, EEG experimental Traub Dose-dependent MA, (2003). M Whittington Danhof & LA, GABA Peletier for PH, 1251-1257. evidence Graaf provide der zolpidem van of FL, Wolters SA, Visser [ M Jenkinson with JD, humans Beaver C, in Salinas receptors S, Tzimopoulou GE, Searle AC, Tziortzi (1999) MA Whittington MA. & Cambridge, JGR, (Oxford, Jefferys psychopharmacology RD, of Traub Journal hyperlocomotion. JM MK-801-elicited Cook 29 or B, England) amphetamine- Markovic not T, but Stankovic 801, P, Biawat of S, of modulation Joksimovic localization T, Autoradiographic Stamenic (1999). 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Spatial ) n=2). = (n A - α 5 Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. Figure 1 F O N O S O N O N O 25 1 Posted on Authorea 16 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159492919.93709869 — This a preprint and has not been peer reviewed. Data may be preliminary. C A Figure

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