Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. diitaino h ihs C oeo 5m/g C oedpnetices nte2aahdnygyeo (2-AG) 2-arachidonoylglycerol the in after increase minutes dose-dependent 30 value BCP baseline A the mg/kg. of 75 85% ex 9.9 to of as with up dose well observed spine) observed was as BCP and plasma levels was in-vitro highest (paw evaluated in hyperalgesia the was tissue evaluated of activity of improvement in and also (MAGL) administration dose-dependent determined 50 lipase were A were Monoacylglycerol (2-arachidonoylglycerol) 25, levels approach. Results. 10, Endocannabinoids evaluated BCP based vehicle, vivo. were HPLC-MS approach. with taken. an (PWR) based using were treated response pain tissues HPLC-MS samples were withdrawal post-surgical and an tissue paw Animals acute dependent using and an model. Time plasma plasma in pain and BCP and (i.p.). of filaments postsurgical intra-peritoneally Frey efficacy acute injected von tested an was we using in that pain, tested acute BCP was in mg/kg BCP BCP 75 for available Methods. are data model. limited Since models. pain Introduction. Abstract 2021 4, July 2 1 Christians Jackson Klawitter Jost of analgesia? mechanism endocannabinoid-mediated new a levels: 2-AG increases β- oeay n os[] h eerhIsiuefrFarneMtras(IM vlae C aey and safety, BCP evaluated sativa, (RIFM) Cannabis Materials of Fragrance oils for essential Institute oil, clove Research especially The plants, [3]. food and hops spice and different rosemary, many of specific oils on essential based the designed was use clinical β- new migraine)[2]. in now with for drugs one of (triptans medications only classes action 10 and four antidepressants of available, than following drugs, mechanisms become fewer the anti-inflammatory have years ketamine; action little non-steroidal 50 and of very opioids, past mechanisms ziconitide, the been treatment: (gabapentin), In has pain antiepileptics prescription [2]. there of (amitriptyline), pain the surprisingly mainstay chronic years However, the of recent been Over field [1]. have the skyrocketed year. in each has innovation Americans or opioids of progress including millions medications affects pain that problem of pervasive a is Pain Introduction activation receptor cannabinoid the 2-AG-mediated Since the rise. propose to we levels 2-AG analgesia. agonist, causing of receptor mechanism in-vivo CB2 BCP’s and and to in-vitro contribute CB1 activity may 7.4 MAGL a of inhibits is IC50 BCP 2-AG an that revealed endocannabinoid showed substrate the data as These 2-AG using Conclusion. assessment activity enzyme MAGL vitro nvriyo ooaoDne colo Medicine School of Medical School Hannover Denver Colorado of University aypyln BP trans- (BCP, Caryophyllene aypyln niismnaygyeo iaeatvt and activity lipase monoacylglycerol inhibits Caryophyllene 1 rsiaSempio Cristina , 1 β- n lbdnM Todorovic M. Slobodan and , aypyln BP a ensont ea ffcieat-namtr gn ncrncadinflammatory and chronic in agent anti-inflammatory effective an be to shown been has (BCP) Caryophyllene 1 ebeWeissenborn Weibke , ± . gm nte7 gk oegopa oprdt eil otoswt 3.0 with controls vehicle to compared as group dose mg/kg 75 the in ng/mL 6.4 β- 1 aypyln)i ao ln oaiecmon on nlreaonsin amounts large in found compound volatile plant major a is Caryophyllene) oj Joksimovic Sonja , 1 aKni Walz MacKenzie , 1 1 1 orjShokati Touraj , 1 eeaKlawitter Jelena , 1 noJust Ingo , μ fBPfrMG inhibition. MAGL for BCP of M 2 1 Uwe , Matthew , ± . gm.In ng/mL. 2.5 Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. emr rn ocrncpi ttswe oprdt ae nbt lncladpelnclsuis[15]; studies preclinical to and considered clinical are both females in (1) males to reasons: main compared two when were for Campus states and study Medical pain Anschutz 1996), our chronic Colorado in Resources, to of rats Animal Guide prone University light-dark female the the more Laboratory h used of with be of We Committee 12 accordance (Institute USA). Use in a Animals CO, and were on months, Care (Aurora, Laboratory protocols maintained Animal (3-5 Experimental of the and rats Use libitum. by cage approved ad Sprague-Dawley and water per Care female and two the young food housed for with to were access performed Animals with were USA). cycle BioCheMed experiments IN, from vivo Indianapolis, preparation in Envigo, standard All calibrator for Animals. used plasma K stripped USA). human charcoal VA, blank (Winchester, and samples USA) USA). QC NY, MI, bury, of Arbor, enrichment (Ann Company the Chemical For Cayman ethanolamide-d3 from stearoyl received and (2-AG- also (PEA-d4) Company 2-arachidonoylglycerol-d5 were ethanolamide-d4 (LEA- palmitoyl (SEA-d4) including Chemical ethanolamide-d4 (OEA-d4), standards linoleoyl Cayman Ethanolamide-d4 (NADA-d8), internal oleoyl from dopamine-d8 labeled d4), N-arachidonoyl received isotope (AEA-d4), were palmitoyl All anandamide-d4 (OEA), USA). d5), (SEA) MI, ethanolamine ethanolamide oleoyl Arbor, stearoyl (LEA), (Ann and ethanolamide (PEA) linoleoyl includ- (OLA), ethanolamide materials N-oleoyl oleamide (O-AEA), reference (ODA), O-arachidonoylethanolamide (2-AGE), dopamine compound ether purchased study 2-arachidonoylglycerol solvent was (NADA), dihomo- All The (PBS0 dopamine (1- (DEA), USA). 1-arachionoylglycerol saline ethanolamide (2-AG), pur- purification. buffered MO, docosatetraenoyl 2-arachionoylglycerol were phosphate AEA), further AG), Louis, study ethanolamide, and without (arachidonoyl this solutions anandamide used (St. in stock ing and Chemicals extraction of USA) sample Sigma-Aldrich preparation NJ, and the from Lawn, phases for (Fair mobile used Scientific for (ethanol) Fisher used from acid) formic chased constituent phase bile reagents and Chemicals Methods of properties anti-inflammatory receptor and CB1 anti-hyperalgesic consequent the and In and to CB2 2-AG contribute receptor. endocannabinoid the could the the CB1) of BCP. of to (and levels levels CB2 bind in endogenous of directly increase the activation the to The to pharmacologically alternative (2-AG). at increases context need 2-arachidonoylglycerol (MAGL) an in consequently the activity agonist show similar lipase without and We monoacylglycerol be activity inhibits concentrations to BCP CB2 [14]. considered relevant that pain the model demonstrate we rodent alters post-operative study a human this BCP is which of which development by 13], the mechanism [12, plantar rats and using to hyperalgesia in mechanisms induced binding paw surgically underlying BCP hind on BCP the direct of of properties no analgesic classification incision belong the or resulting investigated to we weak study, and described this observed mechanism often In controversially as this was which BCP However, BCP studies described [9-11]. CB2, classification. other investigators receptors to postulated this Numerous by cannabinoid BCP been on disputed of receptor). has based recently binding cannabinoid CB2 was agonist possible a receptor CB2 the to cannabinoid putative extensively (binding on well-established the a been family Based its remains with has cannabinoid [8]. action and BCP BCP the al of to of inflammation pain. mechanism been interaction et. and of its has Gertsch Direct however pain treatment BCP by properties, the in [7]. anti-inflammatory for BCP 5]. chronic unknown and candidate cancer, of [4, largely antinociceptive drug to additives efficiency its excellent reference for food The an particular Safetystudied and with BCP Food disorders, make 6]. cosmetic European several toxicity in [4, treating the low used inflammation for by agent be and and promising Administration can pain a Drug which be and agent, to Food flavoring reported the a by as approved Authority been has molecule the . ovnsadraet HL rd ctntie ehnl ae,temo- the water, methanol, acetonitrile, grade (HPLC reagents and Solvents 2 2 DApam a rmBoelmto V (West- IVT Bioreclamation from was plasma EDTA γ- ioeyehnlmd D--E) N-arachidonoyl (DH-g-LEA), linoleoylethanolamide Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. eecnrfgdadspraat eepae noHL il o nlss h ihpromneliquid Supplementary high-performance the The in listed analysis. are section. for parameters Methods vials (HPLC-MS/MS) and HPLC spectrometry Materials into mass tandem placed the – were of chromatography volume supernatants total and The centrifuged minutes. were 20 for temperature room μ 180 a incubated was were mixture samples reaction the kit, the by supplied n 10 and hc a esrduiga PCM prahisedo ooiercraot hspicpewas principle This readout. colorimetric a activity. of MAGL 4- In-vitro instead substrate approach artificial experiments HPLC-MS the Initial an Thus, using applied used. measured assay. was was the MI) which of Arbor, 236 readout of Ann steps colorimetric centration Company, The the Chemical 236 with screen- at Cayman [21]. inhibitor BCP nitrophenylacetate MAGL 705192, used section. of available methods was commercially interference supplementary No. a assay showed the of (Item in published modification detail kit previously A in ing a listed assay. are activity of paw tissue lipase modification and plasma, Monoacylglycerol plasma in A of performed extraction was the tissue. congeners for cord and performed spinal collected. endocannabinoids were 14 and blood site and of tissue incision isoflurane Analysis using the of anesthetized surrounding dose analysis. were tissue second Endocannabinoid animals Any paw a the and received dose analysis. animals cord this further testing spinal after minutes sensitivity drawn, in 30 mechanical was used the solution. was vehicle of response. or completion PWRs a BCP After as threshold considered Each . of not (PWR). collection value was Sample response average animal withdrawal the the paw of for force was and movement the threshold force reaches times voluntary and stimulus a constant other three noticeable, punctate representing and is the tested apparatus withdrawal stand, of paw the was probe pressure the brisk on paw a exerted of immediate displayed the habituation, endure, floor is as After can grams mesh soon animal Animals min. in As the the 15 that through paw. a g. force for the 50 paw utilizes maximum of to habituate the the which 0 area to of mid-plantar from Italy), stand the surface range Varese, mesh to a plantar applied in wire Basile, the paw a (Ugo to the on of apparatus applied enclosures surface Frey was plantar plastic Von the in electronic to placed previously pressure the were exerts described used that was filament we sensitivity animals rigid mechanical single 16 Briefly: assessing and for 6 group 17]. method and dose [16, standard controls) each Our vehicle to Sensitivity. assigned observed 4 Mechanical were randomly and group. differences 50 were group control based (n=8) vehicle mg/kg, mg/kg administration animals the ml/kg (50 volume 25 Eight 2 in No mL/kg in of were mg/kg, group. controls). 4 volume 1/1/18 10 vehicle vehicle the controls), 4 at the saline were vehicle and within i.p. 8 0.9% doses group administered and mg/kg was tested Cremophor/ group (75 solution The ethanol/ vehicle mL/kg mg/kg or 25 of drug 20]. dissolved and solution The (10 [19, post-incision. a mg/kg. protocols h 75 in 2 and published Animals mg/kg dissolved as previous needle. early was as FS-2 with also BCP initiated a accordance and were with elevated experiments suture administration. was all nylon right and muscle Drug 5-0 cages, the plantaris with in of underlying sutured recover surface the was skin to plantar allowed No.11; skin a post- the were were the blade of used and animals whereafter a maintenance) induction we follows: incised, with for the as surgery, longitudinally longitudinally 16-18] and for after [13, incised induction model previously BCP was for standard described paw of (2.5% was Our incision isoflurane effect muscle pain. with antihyperalgesic and post-surgical that anesthetized skin the of believe the study induction we using pain the Thus, to surgical for order [15]. 16]. incision In males [13, muscle using perception and pain model. performed female are pain on animals Incision be in also studies should pain focus current research the of majority the (2) /Lo h nenlsadr rcioi cdd Cya hmclCmay n ro,M) Samples MI). Arbor, Ann Company, Chemical (Cayman acid-d8 arachidonic standard internal the of g/mL μ in-vitro f2A 08 M o150 to mM) (0.85 2-AG of L and μ .Cneunl,teasyraotwsaahdncai A)isedo 4-nitrophenol, of instead (AA) acid arachidonic was readout assay the Consequently, M. in-vivo fe diino 10 of addition After μ μ a usiue ihtentrlsbtae2aahdnygyeo ttecon- the at 2-arachidonoylglycerol substrate natural the with substituted was M .Terato a emntdb diino 800 of addition by terminated was reaction The L. . μ fteMG sa ue 1 MTi-C,p .,1m EDTA) mM 1 7.2, pH Tris-HCl, mM (10 buffer assay MAGL the of L μ AL 10 MAGL, L 3 μ fehnlslto rBPslto nethanol, in solution BCP or solution ethanol of L μ ehnlicuig1.25 including methanol L Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. srto.I nml htudretsi niin efcsdo ehnclhpraagsceet san as effects admin- vehicle hyper-analgesic the mechanical to on compared focused were we i.p. incision, surgery mg/kg paw skin 75 after underwent and postsur- rats 50 that the in 25, on animals effects 10, hyper-analgesic focused In at mechanical we on doses action, istration. BCP escalating of of mechanism in effects potential administered The its when and model. BCP pain of incision gical properties analgesic the assess To Results test. post-hoc compared Tukey’s were with included Groups combination (SEM). statistics mean in Distribution (ANOVA) the of variance errors log-transformation. of standard previous analysis and deviations using without standard analyzed means, of were calculation Data the used. was USA) analysis. data and Statistical 80-120% analyses. of statistical criteria subsequent acceptance all precision generally for following and purpose accuracy for fit with considered curves< guidelines as Calibration validated validation were endocannabinoids. assays biomarker and Both applicable lipids buffer. of surrogate determination in level prepared plasma were internal and the processing data and initial BCP for acquire to Analysis. set Data was spectrometer mass linalool. and -10eV The standard parent to internal -12eV. set was (m/z, was to detector potential linalool potential spectrometry electron buffer standard exit mass The aqueous chromatographic tandem cell mode. the as The (MRM) collision achieve water monitoring to the compounds. reaction in applied endogenous multiple was acid positive from buffer formic in organic linalool operated 0.1% as and using v/v) BCP (1/1, gradient of acid a separation formic and 0.1% PA) with (Sciex, ionization Ford, methanol pressure system Chadds and atmospheric MS/MS positive Inc. API5000 in Analytical operated ABSciex MOD Source an V Ten with Turbo IonDrive mode. interfaced an (APCI) HPLC via series USA) CA, 1200 City, Foster Agilent centrifuged an and using USA) analyzed MA, hundred Waltham, One Scientific, 4 (Fisher USA). and vortexer xg multitube 25,000 a at 160 on and thoroughly briefly mixed vortexed were were Samples added. was solution linalool) μ USA) MA, 5 Waltham, this were Scientific, powder Thirty (Fisher 4 USA). tissue vortexer and MA, of Waltham, xg Multitube 25,000 mg thoroughly at 50 vortexed centrifuged were Approximately and 500 Samples containing pestle. tube noted. and microcentrifuge was mortar pre-weighted 10 a a 0.0025 to using 4, between added nitrogen 1, range liquid 0.4, over the 0.1, homogenized in 0.05, curves 0.02, calibration 0.01, generate to listed used are HPLC parameters into spectrometry section. β- mass placed Methods – were and chromatography Materials supernatants liquid Supplementary and high-performance the centrifuged Five in The were recorded. analysis. Samples 37 was for MI). at tissue vials minutes Arbor, 85 of 60 weight Ann to for Company, added added incubated Chemical the was and and 400 slurry powder scale the of The a to addition added on pestle. were tube and mM) a (0.85 mortar in precooled buffer using assay MAGL nitrogen liquid over homogenized activity. MAGL In-vivo fsml a lcdit . Lmcocnrfg ue hn6 Then tube. centrifuge micro mL 1.5 a into placed was sample of L 0-V epciey eutn eaoiecnetain eenraie otetsu egtadused and weight tissue the to normalized were concentrations metabolite Resulting respectively. 20%-CV, aypyln BP nlss tc ouin fBPwr rprdi tao t1m/L hswas This mg/mL. 1 at ethanol in prepared were BCP of solutions Stock analysis. (BCP) Caryophyllene μ fteitra tnad(1 standard internal the of L μ ehnlicuig1.25 including methanol L netedt eeaqie,Mliun,O-Qvrin17(ce)sfwr a used was software (Sciex) 1.7 version OS-MQ MultiQuant, acquired, were data the Once ° o 0mn(R2icnrfg ihafielt oo,Tem cetfi,Wlhm MA, Waltham, Scientific, Thermo rotor, fiberlite a with centrifuge 23i (MR min 10 for C μ fetatwr netdot aoC nltclclm (2.7 column analytical C8 Halo a onto injected were extract of L μ ftespraatwstaserdit nHL ilwt net ape were Samples insert. with vial HPLC an into transferred was supernatant the of L o nvv ALatvt seset2 o4 go pnlcr isewere tissue cord spinal of mg 41 to 25 assessment activity MAGL in-vivo For μ o ttsia nlssIMSS otaevrin2 IM rok NY, Armonk, (IBM, 27 version software SPSS IBM analysis statistical For ftespraatwstaserdit PCvaswt oia net.To inserts. conical with vials HPLC into transferred was supernatant the of L > agtrin:205 ion): daughter ° μ o 0mn(R2icnrfg ihafielt oo,Tem Scientific, Thermo rotor, fiberlite a with centrifuge 23i (MR min 10 for C μ /L.Frtsu nlsspwtsu n pn isewr manually were tissue spine and tissue paw analysis tissue For g/mL). /Llnlo)slto a de n reyvree.Frpam,40 plasma, For vortexed. briefly and added was solution linalool) g/mL μ /Lo h nenlsadr rcioi cdd (Cayman acid-d5 arachidonic standard internal the of g/mL > 4 4,205 149, μ faeoirl n h cultsu weight tissue actual the and acetonitrile of L > μ 5ad205 and 95 faeoirl a de oec sample, each to added was acetonitrile of L μ /Lt 10 to g/mL μ fteitra tnad(1 standard internal the of L ° .Terato a emntdby terminated was reaction The C. > 5frBP 137 BCP; for 55 μ /L(,002,0.005, 0.0025, (0, g/mL μ ,3010m MAC- 3.0x100mm, m, μ f2-AG-d8 of L > 1frthe for 81 μ g/mL μ of L Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. gk ramn rus respectively. showed groups, controls treatment Vehicle BCP. mg/kg with Formation treated S1). previously Figure 3.76 animals pg supplementary of activity of and rate and tissues MAGL extracted, formation section cord with was AA-d8 spinal methods tissue slurry an in see the a reduced details was period, into (for AA-d8 incubation BCP placed analyzed of the after were was After minutes content 2-AG-d8. samples 30 AA-d8 labeled tissue, harvested the with in was -70 incubated Tissue acid and at arachidonic buffer mg/kg. stored assay at and 75 and BCP and 2-AG with nitrogen 50 of treated 25, liquid abundance were 10, in Animals as frozen well tissues. cord as administration, controls), spinal (vehicle in mg/kg assessed 0 also was activity 0.9445. lipase = the Monoacylglycerol conditions r these was Under fit day. per curve assessments the 2 (see natu- for with was parameters days the assay (AA), 4 which of consecutive acid on in optimization IC arachidonic applied After developed was product, S1). was assay the Figure the assay supplementary and methods) the (see used with of HPLC-MS/MS was BCP using modification (2-AG) of quantified 2-arachidonoylglycerol a interference monoa- showed Thus, substrate on assay MAGL (Cayman 3A). BCP the kit ral of (Figure application, assay effect this readout For screening an colorimetric 3A). by inhibitor the (Figure caused MAGL tested available was initially concentrations commercially was 2-AG a Chemicals) activity, of observed (MAGL) increase was the lipase 0.518 tissues. that cylglycerol = paw hypothesis r in (Pear- our coefficient observed assessed test correlation was To was a correlation concentrations with such However, BCP 0.0024) No groups. and = S2). study 2-AG (p figure between correlation tissue (supplementary changes significant cord significant a the no spinal Correlation), in with between son resulted levels association tissue plasma potential 9.9 cord 2-AG a with spinal increased when and group showed tissue mg/kg groups paw 3.0 dose treatment 75 in were other BCP 2-AG the group all mg/kg control for group 75 vehicle value mg/kg the the highest 10 for in levels the plasma endocannabinoids plasma of 14 in 2-AG exception the levels concentrati- Mean Amongst 2-AG 2-AG 3-fold. 2). increased plasma about (Figure significantly that group administration statistically noticed BCP endo- observed We increasing in we samples. with changes analyzed tissue parallel evaluated 14 and in including we activity, increased analysis plasma ons endocannabinoid analgesic in comprehensive BCP congeners a of and using mechanism concentrations endocannabinoids the tissue paws and into affected plasma insight for cannabinoid gain analgesic further selective relatively to a Next, is injection BCP i.p. incision. that post indicates minutes skin 15 This surgical at control. following On group vehicle mg/kg side). the 75 right 42.0 to the 1 of compared in average PWR (Figure in an animals increase showed study transient (38.5 paws (51.7 and of values left moderate hind-paws the pre-surgery a assessments left for the PWR non-incised Except than the the lower of in day 15% impro- evaluated the only PWR also were showing were (n=8) PWR group group The this treatment for injection. BCP responses post mg/kg 30-minutes withdrawal 75 and the 31.4 15- for with at pronounced 10.5 vements responses most of withdrawal were paw average changes diminished an showed These 38.5 at animals was remained treated analgesic incision 9.7 BCP (n=16) selective the to all group a noticed to dropped vehicle We value indicating prior the this paws paws panels). While post-surgery operated right right hours in 1 the 2 PWRs (Figure for At paws the PWR deviation). (left) of baseline analgesic non-operated increase The (hyper and dose-dependent incised effect. biochemical side) the a and left in setting caused pharmacological determined 1 clinical BCP to were (Figure a that sensitive (PWR) paw in is responses hind observed it withdrawal right commonly previously, paw very published effect) The as is 17]. and, it [16, 23] reasons: 22, modulation main [16, two pain for incisional pain of incisional of feature important · 50 mg o C a eemndt e7.4 be to determined was BCP for ± -1 · min . rm) hr een infiatcagsi h etpwPRaogtaltetetgroup treatment all amongst PWR paw left the in changes significant no were there grams), 6.6 -1 3.16 , ± ± . n 32.9 and 4.1 .7pg 0.57 · mg ± -1 ± .6pg 0.96 · min . rm o 0ad6 iue otijcin epciey h paw The respectively. injection, post minutes 60 and 30 for grams 3.5 -1 μ 9%cndneitra . o1.) h orlto coefficient correlation The 14.0). to 3.9 interval confidence (95% M 2.99 , · mg -1 ± · min ± .2ad2.58 and 1.12 . gm (p ng/mL 6.4 -1 5 h ausfrtetetetgop ee3.56 were groups treatment the for values The . º rblwutlaayi.Det h natural the to Due analysis. until below or C ± ± . rm niaighpraagsceffects. analgesic hyper indicating grams 4.5 < ± .3( .3)frte1,2,5 n 75 and 50 25, 10, the for 0.036) = (p 0.73 .0) bouetsu ocnrtosof concentrations tissue Absolute 0.001). . rm uigtetetetperiod, treatment the during grams 2.0 ± . rm o h eil group. vehicle the for grams 4.0 ± . rm (mean grams 4.5 ± . gm.Wt the With ng/mL. 2.6 ± ± . grams). 4.5 standard ± 1.01 Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. t1,3,6 n 0mntsatrijcincmae ovhcecnrl 10) diitaino 75 of Administration (32.9 (100%). injection controls post vehicle minutes to (145-167%) 30 compared PWRs at the injection higher responses significantly to after withdrawal showing contributing minutes paw mg/kg) effect 90 in (10 anti-inflammatory and post-operative resulted tested additional of 60 mg/kg dose model an 30, dependent BCP relevant exclude 15, dose lowest clinically in cannot at BCP The this BCP we a using of observed However, effect. an paws effect have in analgesic operated We antinociceptive BCP 1). for in the of studied. mg/kg/day (Figure preferentially tested effects previously 2.6 pain anti-hyperalgesic PWR models been [19], the the these not knowledge mice of of has our in increase To Most model mg/kg tests. pain [41]. 1-5 immersion post-surgical mg/kg at tail acute 25 and antinociceptive This BCP. and acid, be of acetic [42] [29]. to effect formalin, mice showed compound the in BCP to active weeks context components major 2 oil this a essential In is displays other mg/kg BCP BCP designs. of 400 pure which contribution - surprisingly, and in 100 and bioavailability oils variable and oils, Pure to essential [37], often essential related rich mg/kg is several be BCP 75 oils as might essential - 36-41]. activities 50 in [19, content [40], analgesic described BCP mg/kg effect similar well However, 62.5 analgesic been - The (p.o.). has 25 39] 31-35]. formulations at [36, [7, other effects cells antinociceptive and glial demonstrated oils and have neurons essential neuro-inflammation, cerebellar in lesions, and BCP brain hippocampal, of ischemia, to cortical, cerebral potential as in the such problems has disorders and and BCP diseases 28-30]. neurological activities several [7, pharmacological treat neuroprotective important anti- and displays antioxidant, immune-modulatory, also nephroprotective, antimicrobial, gastroprotective, It inflammatory, hepatoprotective; [28]. cardioprotective, agent anticancer, flavoring as and such both enhancer, to taste agonistically additive, acts 2-AG food as receptors CB2- inhibition BCP. β- to – of BCP mechanism of proposed agonism selectivity anti-inflammatory the CB2-receptors. the However, CB2 and and abolishing action. anti-hyperalgesic putative CB1- in of results the the mechanism – this as MAGL to via such of contribute explained BCP, 2-arachidonoylglycerol. be may of agonist could we lipase (CB2) properties, properties study 2 monoacylglycerol characteristic this receptor In many of [9-11]. Cannabinoid However,Importantly, weak inhibition the rather agonist. of the is CB2 whether levels interaction controversial, Hence, the the putative remains if increased it a or 24-27], consequently agonist considered [8, and an others been as and acts al. has that and et showed CB2 BCP Gertsch to from [8] directly evidence binds of al. BCP lot et a is Gertsch there of although findings the Following Discussion 5A-5C). (Figure respectively groups, treatment mg/kg 75 and 50 25, opam n o4fl ihri oprsnt a isedpnigo h ramn ru.Sia cord Spinal group. treatment the on depending tissue paw comparison to in comparison higher 18.61 in 9-fold reached higher to concentrations 4-fold 8 BCP to even 3 were and concentrations plasma cord to spinal the Interestingly, respectively. groups, 0ad7 gk ramn rus epciey a iselvl eehge hnpam eeswt 6.16 with levels plasma than higher were com- 2.34 levels easy were tissue for levels Paw respectively. plasma allow groups, BCP to the treatment 5A-5C). ± to tissue mg/kg Figure tissue normalized of 75 (see and were and nmol/g nmol/mL plasma levels 50 as in All Tissue presented presented assay. nmol. are are the which 490 ± and level, of to acetonitrile plasma range 0.49 with to linear from parison extracted the linear tissue within was of were assay samples amount animals The plasma A treated in S3. ad- BCP levels. of Figure BCP second of samples quantify in-vivo Supplementary chromatograms a to to in ion developed after extracted shown data was Representative assay minutes is selectivity. in-vitro multi- (MS/MS) and 30 correlate spectrometry specificity (APCI) mass high tissue and ionization tandem with chemical paw levels (MRM) pressure tissue monitoring and reaction atmospheric and cord ple (HPLC) plasma spinal chromatography estimate liquid plasma, to high-performance in BCP determined of were ministration levels drug BCP aypyln sapoe yteUie ttsFo n rgAmnsrto n uoenaece sa as agencies European and Administration Drug and Food States United the by approved is Caryophyllene .4 4.60 1.34, .2 9.73 2.22, β- aypyln a entse tvroscnetain nsnl n utpeamnsrto dose administration multiple and single in concentrations various at tested been has caryophyllene β- ± ± aypyln niismnaygyeo iaea hraooial eeatconcentrations relevant pharmacologically at lipase monoacylglycerol inhibits caryophyllene .3 5.48 2.33, .6 14.68 3.56, ± ± .8ad15.90 and 1.88 .8ad40.37 and 6.38 ± .4 39.13 6.94, ± ± .4no/L(mean nmol/mL 4.84 ± 06 mlgfrte1,2,5 n 5m/gtreatment mg/kg 75 and 50 25, 10, the for nmol/g 10.62 37,50.38 13.78, 6 ± 50,133.85 25.09, ± tnaddvain o h 0 25, 10, the for deviation) standard ± ± 29 mlgfrte10, the for nmol/g 22.92 . rm)ta were that grams) 3.5 Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. eedn .- or fe diitaindpnigo h eil ouin sd ntepeetstudy present the analysis In pharmacokinetic used. C performed solutions plasma determined vehicle rat Others and the in 53] observed on dose. mg/kg [52, nmol/mL depending 10 BCP rats 2.3 administration of in second to after administration BCP compared the hours mg/kg dose [51] 1.5-3 after 50 single nmol/mL depending minutes of after 1.2 administration 30 minutes with ca. oral alignment after 30 of after explained in determined study levels generally be serum this are can al. mice observed which in et BCP in plasma, Varga of to i.p. and concentrations compared increased 52]. BCP The 50 were as BCP. levels [51, 25, cord BCP of literature spinal 10, nature well. ( in the the as lipophilic accumulated nmol/mL levels for very further tissue 15.90 the linear even cord 5.48, by spinal not and 4.60, and tissue is paw 2.34, dependent paw increase in of concentration in reflected the values a was However, mean show increase other level with did interpret to 5). non-linear groups study administration accurately (Figure this treatment second to from groups mg/kg a levels gained study 75 after data tissue the minutes of plasma comparison 30 which amongst determine a BCP, levels increase to for of BCP allowing essential formulations Plasma as is well and studies. it as administration experiments terpene, of in-vivo volatile routes and in-vitro and different the chromatography lipophilic liquid to a performance Due high is by spectrometry. determined been mass have concentrations tandem BCP tissue cord spinal indicates IC and also BCP’s vivo if ex tissue in-vitro determine tissue of To the of incubations than that 2-AG higher to labeled much Also identical are be spine) group. inhibition. increased treatment not and MAGL the (paw might BCP-induced effects on tissue assay the anti-inflammatory depending in inhibitor value, that describe found MAGL IC50 concentrations well posit in-vitro that BCP and the we established the antinociceptive in However, Therefore, well observed determined MAGL. is [50]. IC50 the [46-49]. plasma The agonist It to BCP. effects contribute in CB2 or anti-inflammatory could manner. a levels and 2-AG and dependent 2-AG antinociceptive of CB1 dose an increased levels a BCP have a BCP is that levels in why 2-AG demonstrate 2-AG S2) Importantly, explains increased data Figure indicating and likely These (supplemental animals inhibition most 4). tissue treated MAGL and (Figure cord mg/kg in-vivo group 75 spinal treatment and from and significant this tissue in-vitro statistically in samples cord MAGL A 2-AG-d8 (2-AG-d8, slurry spinal inhibits HPLC-MS. of tissue 2-AG in using hydrolysis the observed deuterated determined of decreased was this was incubation a formation For AA-d8 After AA-d8 product of substrate. groups. MAGL MAGL decrease treatment the the and BCP as extracted the used were was from S1) animals Figure of Supplementary 2-oleoylglycerol homogenates utilizing tissue mostly sub- 45] in natural [44, the others IC (AA) by using the The acid applied arachidonic applied with been product was substrate. BCP catalytic has the strategy of the as methodology modified analyzing interference (2-OG) similar and a to A assessment 3A) due activity HPLC-MS. (Figure However, MAGL by was activity. the kit via assay for MAGL assay levels screening 2-AG, 2-AG inhibit the inhibitor strate, increases MAGL to of BCP available ability readout that commercially BCP’s hypothesize colorimetric a the evaluate hypothesis to this to us test used led To treat- results can MAGL. BCP specifically These of of endocannabinoids, agonists) 2). effects circulating inhibition receptor in The (Figure hydrolase changes CB2 [43]. observed and amide levels evaluated were or was neurophysiological acid 2-AG CB1 system these fatty endocannabinoid (e.g. of with the three ligands on (e.g. all ment endocannabino- cannabinoid cannabinoids at the exogenous transmission regions endogenous targeting nociceptive supraspinal or result, of reduce multiple a inhibitors) levels in As MAGL the and [43]. of or cord modulation (FAAH) enhancement spinal and the via perception of pain system horn with peri- id dorsal the associated the the in brain pathways: in of the nociceptive neurons, characteristics of throughout afferent the expressed primary are on on system based effective phery endocannabinoid likely the the is of to components tests while difference Key immersion The BCP BCP. tail of of and effect dose models. acid analgesic highest pain formalin/acetic (38.5 different the postoperative mice incision with markedly the the affected in a slightly to doses show only prior data were values paws These baseline BCP. non-operated all of for effect PWR anti-hyperalgesic mean the of 85% 50 o C a eemndt e7.4 be to determined was BCP for 50 f7.4 of μ a erahdi-ioatrBPamnsrto lsa a tissue paw plasma, administration BCP after in-vivo reached be can M μ .Asmlrapoc a ple ots h ALactivity MAGL the test to applied was approach similar A M. 7 ± . rm)idctn potent a indicating grams) 4.5 max ees25–1 nmol/mL 10 – 2.5 levels μ ) epciey This respectively. M), Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. npam A,pwtsu B n pnlcr ise()aeson aaaesona en+standard + mean as shown are Data shown. are (C) tissue cord concentrations spinal BCP and administration. (B) BCP second tissue after paw minutes (A), 30 plasma tissue and in plasma in concentrations BCP 5 Figure mean as shown test. are concentrations HSD Data Tukey’s the AA-d8 standard. AA-d8. using internal (p and significantly as performed statistically glycerol AA-d5 was indicates using to evaluation MAGL HPLC-MS/MS activity by using MAGL hydrolyzed determined tissue. were is which cord 2-AG-d8, spinal substrate in deuterated activity MAGL of Determination hydrolyzed is which HPLC-MS/MS. 2-AG, using 4 determined substrate Figure were natural concentrations mean the AA as (AA). on shown acid based are arachidonic Data BCP and glycerol by 18,000 hydrolyze to of inhibition MAGL to coefficient MAGL by MAGL extinction (B) of specific ability readout. a the metric has on which based 4-nitrophenol, is and M evaluation acetate activity form The available to commercially kit. 4-nitrophenylacetate the assay using evaluation screening activity inhibitor inhibitor MAGL MAGL (A) inhibition. MAGL of Determination en- based HPLC-MS/MS test. HSD a 3 Tukey Figure of post-hoc part on (p as based significantly determined group statistically control were indicates vehicle levels *** assay. 2-AG multiplex levels. docannabinoid plasma 2-AG of Determination higher significantly left statistically showed non-incised 2 groups the Figure treatment for all PWR administration, corresponding BCP the mean after side, as (p minutes shown with Right are 90 groups Data mg/kg. and study incised 75 groups. the 60 the study and for for same 50 and the PWR of 25, (black) side, paws group 10, Left study at testing. control groups filament vehicle treatment Frey the von for using paw observed right (PWR) response withdrawal Paw 1 Figure However, targeted Legends and effects. selective Figure more side receptors a increased all be in might of results inhibition activation might MAGL in density, by results receptor concentrations weak agonists 2-AG tissue rather approach. local CB2 on observed is of or was depending effects interaction modulation anti-inflammatory CB1 it and the the and of recently body, if antinociceptive However, Administration of the the or levels to in agonist increased 24-27]. administration. contribute consequently, an least [8, and, BCP as inhibition at agonist with MAGL analgesic, acts receptor that known and suggest CB2 a CB2 strongly 2-AG, putative with data to a injections our directly Alternatively, is i.p. binds [9-11]. twice BCP BCP of that whether kinetics proposed the disputed of been comparison model has a this It Thus, In excellent possible. place concentrations. concentration. not to takes plasma state is due redistribution in steady administration Afterwards concentrations increase a oral an at CNS brain/spine. and not high the concentrations are very of brain/spine we in lipophilicity of lowering result high in of to resulting combination injection ( in nmol/mL supply i.p. by 5.5 blood administered of drugs levels lipophilic plasma average observed we -1 < cm .1 enPR oprdt h eil oto group. control vehicle the to compared PWRs mean 0.01) -1 t45n.Tedse raidctsitreec fBPa ihcnetain ihtecolori- the with concentrations high at BCP of interference indicates area dashed The nm. 405 at ± tnaderro h en(SEM). mean the of error standard < .5 hne oprdt h eil oto ru ae npost-hoc on based group control vehicle the to compared changed 0.05) 8 μ )i h 0m/ggop ti nw rmvery from known is It group. mg/kg 50 the in M) ± tnaddvain o h nie a t1,30, 15, at paw incised the For deviation. standard < .0)cagdcmae othe to compared changed 0.001) β- aypyln (BCP) caryophyllene ± E.* SEM. Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. iue3 Figure 2 Figure for 1 6), Figure = (n group mg/kg 50 the for Figures levels difficulties. plasma technical the of of because exception available the not were with samples 8, plasma = 2 n which are data All deviation. 9 Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. iue4 Figure 10 Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. iue5 Figure 11 Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. 12 Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. oietr n leitspssria pain. postsurgical alleviates al., and et nociceptors S.L., Joksimovic, pain.18. post-surgical of model rodent 1735-1753. a in p. analgesia al., effective Channels. et exerts Calcium T-Type properties S.L., of Joksimovic, Role the 17. for Evidence Pain: Post-Surgical al., of Model et Rodent Q.L., Tat, 16. 5 oi,JS n ..Chanda, M.L. and pain. J.S. Mogil, Schug, 15. S.A. and Segelcke, 2017. D. Rep, Pain E.M., Pogatzki-Zahn, incision. paw 14. surgical of model 48-61. rat p. a (1): in epipregnanolone by analgesia al., mediated et S.L., Joksimovic, 13. Gebhart, G.F. 1996. and Vandermeulen, Pain, E.P. T.J., Brennan, 12. Channels. TRPV1 and TRPA1 on 2020. Endocannabinoids al., or Phytocannabinoids et of M., Receptors. Heblinski, CB2 and 11. CB1 Human at 2019. Delta(9)-THC Res, of noid Activity Functional al., the et Modulate M., Santiago, 10. Receptors. noid al., et D.B., Finlay, 9. 9099-104. al., p. et (26): J., Gertsch, 8. 2018. al., Res, et Phytother K.D.C., Machado, 7. 2019. al., Sciences, et F., Francomano, Carroll, 6. B. and Levy, Rats. R. in Toxicity D., Schmitt, 5. Literature. Published of Analysis titative M.E., Maffei, 4. al., et C., Ghelardini, 3. 7. p. D., Borsook, 2. format) specific 2 no J., Trecki, 60, 1. than more no (ideally References 1:p e187104. p. (1): an 2005. Pain, 5 4:p 305-317. p. (4): 64 esetv eadn h urn tt fteOii Epidemic. Opioid the of State Current the Regarding Perspective A 3:p 493-502. p. (3): 9 2 uuewtotcrncpi:nuocec n lnclresearch. clinical and neuroscience pain: chronic without future A 4 rn hrao,2020. Pharmacol, Front ln aua ore fteEdcnaiod()bt-aypyln:ASseai Quan- Systematic A (E)-beta-Caryophyllene: Endocannabinoid the of Sources Natural Plant 2) .5420. p. (24): 117 n oio,2016. Toxicol, J Int 2:p e588. p. (2): 3:p 165-176. p. (3): 12:p 1-5. p. (1-2): 32 epnisFo anbsD o eit nEtuaeEetb ciga Cannabi- at Acting by Effect Entourage an Mediate Not Do Cannabis From Terpenoids eacrohleei itr cannabinoid. dietary a is Beta-caryophyllene repieAagscEeto nrtea plctoso eratv trisi a in Steroids Neuroactive of Applications Intrathecal of Effect Analgesic Preemptive oa netei ciiyo beta-caryophyllene. of activity anaesthetic Local 1) .2376-2388. p. (12): bec fEtuae epnisCmol on nCnai aiaD Not Do sativa Cannabis in Found Commonly Terpenoids Entourage: of Absence -ροηλεε εχιεπν ιηὃνλσ ιλγςλΠροπερτιες. Βιολογιςαλ ὃυντλεσς ωιτη Σεσχυιτερπενε Α β-ἃρψοπηψλλενε: epnisCmol on nCnai aiaD o ouaeteActions the Modulate Not Do sativa Cannabis in Found Commonly Terpenoids niiino utpevlaegtdclimcanl a otiuet spinally to contribute may channels calcium voltage-gated multiple of Inhibition eetv niiino a32canl eesshprxiaiiyo peripheral of hyperexcitability reverses channels CaV3.2 of inhibition Selective ytmtcrve ntenuortcieprpcie fbeta-caryophyllene. of perspectives neuroprotective the on review systematic A oe eratv tri ihhpoi n -yeclimcanlblocking channel calcium T-type and hypnotic with steroid neuroactive Novel h aefrteicuino eaesbet nbscsinesuisof studies science basic in subjects female of inclusion the for case The 35 11 n o c,2020. Sci, Mol J Int 5:p 558-67. p. (5): oiooia vlaino eaCrohleeOl Subchronic Oil: beta-Caryophyllene of Evaluation Toxicological .359. p. : c inl 2018. Signal, Sci 13 otprtv anfo ehnsst treatment. to mechanisms pain-from Postoperative hrceiaino a oe ficsoa pain. incisional of model rat a of Characterization 11 21 (545). (18). rcNt cdSiUSA 2008. A, S U Sci Acad Natl Proc amc,2001. Farmaco, rJPamcl 2020. Pharmacol, J Br hnes(utn,2019. (Austin), Channels anbsCnaiodRes, Cannabinoid Cannabis AANt pn 2019. Open, Netw JAMA eerm 2012. Cerebrum, anbsCannabi- Cannabis 56 el,2020. Cells, 57:p 387-9. p. (5-7): 177 Applied 9 2012 (12). 105 (8): 13 : Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. il 2010. al., Biol, et D., Mishra, 37. al., leaves. et erythropappus O.V., Sousa, 36. fascicularis. Macaca al., of et J.L., Lanciego, 35. 2009. R.G., Pharmacol, Pertwee, 34. al., et injury. chemic/reperfusion M., Zhang, 33. neuroprotection. to concomitant 211-9. activation al., macrophage/microglial et brain alternative J.G., Zarruk, 32. injury. ischemic/reperfusion cerebral al., during et M., Zhang, 31. al., Cells. Glioblastoma et in N., Receptors Irrera, 26. Imaging. al., Cell et neurodegeneration. Live towards Based L., tool Mensching, strategic 2021. a 25. Appl, as Biol agonist Mater CB2 C cannabinoid Eng Sci of Maraschin, delivery M. and drug Coelho, Controlled D.S. T.B., Alberti, 24. T.J., Brennan, 23. Brennan, pain. T.J. and P.K. chromatography- Zahn, liquid 22. ionization chemical pressure spectrometry. atmospheric mass high-performance switching al., column et using C., Sempio, 21. Ischemia. Permanent of al., Model et Mouse C.G., Yokubaitis, 20. pain. neuropathic and inflammatory of models 24 mouse in effects analgesic al., et A.L., Klauke, 19. itr htcnaiodo hraetclPromise. Pharmaceutical of Phytocannabinoid al., et Dietary C., Sharma, 30. al., properties. et analgesic K., Fidyt, 29. al., et A., system. endocannabinoid Chicca, Receptors. PPAR-gamma 28. and CB2 al., between Cross-Talk et a N., Through Irrera, 27. 4:p 608-20. p. (4): nshsooy 1999. Anesthesiology, 48 1) .1297-301. p. (11): 156 ahpyilg fpsoeaiepain. postoperative of Pathophysiology nlBonlCe,2021. Chem, Bioanal Anal mrigsrtge o xliigcnaiodrcpo gnssa medicines. as agonists receptor cannabinoid exploiting for strategies Emerging acrMd 2016. Med, Cancer eacrohleeadbt-aypyln xd-aua opud fatcne and anticancer of compounds oxide-natural beta-caryophyllene and beta-caryophyllene hr hrao,2008. Pharmacol, Pharm J hmclcmoiinadaagscatvt fSncorfievsesniloil. essential rufinervis Senecio of activity analgesic and composition Chemical ouaino h aac ewe anbni B1 n B2 eetractivation receptor CB(2) and CB(1) cannabinoid between balance the of Modulation scohrao,2011. Psychopharmacol, J 3:p 397-411. p. (3): eaCrohleeMtgtsClae nioyIdcdAtrts(AA nMice in (CAIA) Arthritis Induced Antibody Collagen Mitigates beta-Caryophyllene eaCrohleeIhbt elPoieaintruhaDrc ouaino CB2 of Modulation Direct a through Proliferation Cell Inhibits beta-Caryophyllene oyhraooia rprisadTeaetcPtnilo eaCrohlee A beta-Caryophyllene: of Potential Therapeutic and Properties Polypharmacological B eetratvto teutsmcoicltr yfnto uigcrba is- cerebral during dysfunction microcirculatory attenuates activation receptor CB2 nlsso 4edcnaiod n noanbni ognr nhmnplasma human in congeners endocannabinoid and endocannabinoids 14 of Analysis h anbni B2 eetrslciepyoanbni eacrohleeexerts beta-caryophyllene phytocannabinoid receptor-selective CB(2) cannabinoid The nioietv n niiflmaoyeet fteesnilolfo Eremanthus from oil essential the of effects anti-inflammatory and Antinociceptive ucinlzto fbt-aypyln eeae oe oyhraooyi the in polypharmacology novel generates beta-caryophyllene of Functionalization C hmBo,2014. Biol, Chem ACS xrsino h RAcdn h anbni eetr2i h aldlcomplex pallidal the in 2 receptor cannabinoid the coding mRNA the of Expression anbni ye2rcpo ciaindwrgltssrk-nue lsi and classic stroke-induced downregulates activation receptor 2 type Cannabinoid n o c,2020. Sci, Mol J Int 90 oioigCnaiodC2-eetrMdae APDnmc yFRET- by Dynamics cAMP Mediated -Receptor CB2 Cannabinoid Monitoring irvs e,2009. Res, Microvasc ffcso anbdo n eaCrohleeAoeo nCmiaini a in Combination in or Alone Beta-Caryophyllene and Cannabidiol of Effects 3:p 863-72. p. (3): acr Bsl,2020. (Basel), Cancers rmr n eodr yeagsai a oe o ua postoperative human for model rat a in hyperalgesia secondary and Primary 121 .111824. p. : n.J o.Si,2021. Sci., Mol. J. Int. 5 1) .3007-3017. p. (10): ersine 2008. Neuroscience, 413 21 9 78 25 eaCrohleennprilsdsg n development: and design nanoparticles beta-Caryophyllene 60 (21). 1) .3381-3392. p. (12): 7:p 1499-507. p. (7): 14 1:p 86-94. p. (1): 1:p 97-104. p. (1): 6:p 771-7. p. (6): urPamDs 2016. Des, Pharm Curr 12 an 2011. Pain, (4). 22 152 26) .1-11. p. (2866): imlcls 2019. Biomolecules, 3:p 753-60. p. (3): 152 u erpyhpamcl 2014. Neuropsychopharmacol, Eur 3Spl:p S33-S40. p. Suppl): (3 22 2) .3237-64. p. (21): toe 2012. Stroke, 9 (8). 43 Pharm 1:p. (1): Mater rJ Br Posted on Authorea 4 Jul 2021 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.162539258.80061327/v1 — This a preprint and has not been peer reviewed. Data may be preliminary. ieDfiisi aswt aclrDmni hog h anbni eetrTp Mdae Pathway. -Mediated 2 Type Receptor Cannabinoid the through Dementia 2017. Vascular Pharmacol, with Front Rats in al., Deficits al., et tive J., et Lou, 53. H., Liu, complex. inclusion caryophyllene/beta-cyclodextrin 52. mice. in dysregulation metabolic al., and et mation Z.V., Stelt, Varga, der van 51. brain. M. the and in Maccarrone, actions M. manifold M.P., Baggelaar, 50. 2019. al., Pharmacother, et J., Sun, 49. vivo. in al., crosstalk et cannabinoid J.Z., Long, 48. mice. al., in et effects cannabimimetic B., Ignatowska-Jankowska, 47. effects. gastroprotective and 2013. antinociceptive 1-mediated Ther, type receptor al., cannabinoid et retains S.G., Kinsey, 46. 157-68. Piomelli, p. D. : and K.M. Jung, 45. slices. brain intact in al., degradation et A.R., King, 44. 2021. al., Pain, et D.P., Finn, 43. al., Acid. docosahexaenoic et and P., caryophyllene Fiorenzani, 42. mice. al., in et G.C., Segat, 41. oil. al., essential et castus E., Khalilzadeh, 40. al., et mice. I.A., Menezes, Oil. 39. Essential Cav. lucida al., Tagetes of et Effects A., Hernandez-Leon, 38. ioeai,2007. Fitoterapia, erpamclg,2017. Neuropharmacology, 345 3:p 492-501. p. (3): h noanbni ytm oe agt o raigcne nue oepain. bone induced cancer treating for targets Novel system: endocannabinoid The eaCrohleeHdoyrplbt-yldxrnIcuinCmlxIpoe Cogni- Improves Complex Inclusion beta-Caryophyllene/Hydroxypropyl-beta-Cyclodextrin vcnaJPyoe,2015. Phytomed, J Avicenna 120 ulbokd fFA n ALietfisbhvoa rcse euae yendo- by regulated processes behavioral identifies MAGL and FAAH of blockade Dual anbnis h noanbni ytm n an eiwo rciia studies. preclinical of review a pain: and system, endocannabinoid the Cannabinoids, R62ihbt oocllcrllps n eetvl lcs2-arachidonoylglycerol blocks selectively and lipase monoacylglycerol inhibits URB602 niloyi ffc fbt-aypyln npciae-nue eihrlneuropathy peripheral paclitaxel-induced on beta-caryophyllene of effect Antiallodynic eaCrohleepoet gis looi taoeaii yatnaiginflam- attenuating by steatohepatitis alcoholic against protects beta-Caryophyllene 8 78 eetdlwds diitaino h oocllcrllps niio JZL184 inhibitor lipase monoacylglycerol the of administration low-dose Repeated .109504. p. : nioietv ffc n ct oiiyo h seta i fHpi rtcs in fruticosa Hyptis of oil essential the of toxicity acute and effect Antinociceptive .2. p. : hscceia hrceiainadpamckntc vlaino beta- of evaluation pharmacokinetics and characterization Physicochemical nioietv ffcs ct oiiyadceia opsto fVtxagnus- Vitex of composition chemical and toxicity acute effects, Antinociceptive nvtoadi iocaatrzto ftenwaagsccmiainBeta- combination analgesic new the of characterization vivo in and vitro In 3:p 192-5. p. (3): rcNt cdSiUSA 2009. A, S U Sci Acad Natl Proc rgLpdRs 2018. Res, Lipid Prog hrao x hr 2015. Ther, Exp Pharmacol J oeo eaCrohleei h nioietv n Anti-Inflammatory and Antinociceptive the in beta-Caryophyllene of Role sa fMnaygyeo iaeActivity. Lipase Monoacylglycerol of Assay hmBo,2007. Biol, Chem 125 eetv oocllcrllps niios nioietv versus antinociceptive inhibitors: lipase monoacylglycerol Selective .207-219. p. : vdBsdCmlmn lentMd 2014. Med, Alternat Complement Based Evid rJPamcl 2018. Pharmacol, J Br oeue,2020. Molecules, 5 3:p 218-30. p. (3): n hr,2013. Pharm, J Int 15 14 71 1) .1357-65. p. (12): .1-17. p. : -rcioollcrl inln ii with lipid signaling A 2-Arachidonoylglycerol: 25 353 106 (3). 2:p 424-32. p. (2): 4) .20270-5. p. (48): 175 450 2:p 320-334. p. (2): 12:p 304-10. p. (1-2): ehd o il 2016. Biol, Mol Methods 2014 hrao Exp Pharmacol J .596312. p. : Biomed 1412