Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. adn Yi Yaodong dorsal the ganglion in root uptake transporter-mediated the inhibiting selectively by neurotoxicity oxaliplatin-induced reduces L-tetrahydropalmatine X-nue hoi eihrlnuooiiymc n uorbaignd iewr used were utilized mice were nude lines tumour-bearing cell and tumour mice neurotoxicity and peripheral cells chronic DRG OXA-induced primary cells, and MDCK OXA Transporter-transfected of and transport transporters transcellular uptake Approaches the the Experimental in inhibit OCTN2 neurotoxicity. selectively This and peripheral would application. OXA-induced OCTN1, (L-THP) its alleviate OCT2, L-tetrahydropalmatine subsequently limits of which whether role neurotoxicity, explore peripheral the to severe confirm induces OXA to of (DRG) aims accumulation ganglia high study root the however, dorsal drug; the platinum anti-tumour in third-generation a is (OXA) Oxaliplatin Purpose a and is Background L-THP OXA. of efficacy neurotoxicity. antitumour Conclusion peripheral the OXA-induced cells). impairing attenuate SW620 without OXA-induced to attenuates and but L-THP drug OCTN1 (HT29 DRG. candidate and the cells potential in OCT2 tumour uptake impair inhibiting OXA in not to via did nor contribute neurotoxicity and mice OCT2, tumours peripheral nude especially the MDCK-hMRP2 OCTN1, in tumour-bearing and concentration from HT29 the OCT2 platinum Implications in efflux in the and played reduce OXA concentration its not of concentration-dependent OCT2 platinum did affect efficacy OXA; L-THP the a not antitumour reduced of manner. in the did dose-dependent and uptake OXA a but neurotoxicity the in of cells peripheral mice in accumulation in OXA-induced DRG involved DRG cellular attenuated primary were L-THP and rat OCTN2 Furthermore, cytotoxicity were and not the mice cells. MDCK-hOCTN1, but whether nude reduced MDCK-hOCT2, OCTN1 tumour-bearing L-THP explore in and and to manner OCT2 role. mice and Results neurotoxicity important OXA Key peripheral most chronic cell of the studies. OXA-induced tumour transport vivo OXA-induced and studies. study in transcellular cells alleviate vitro DRG This used the subsequently in primary in and utilized cells, application. were MDCK OCTN2 transporters its Transporter-transfected lines uptake and limits Approaches which the Experimental OCTN1, neurotoxicity, neurotoxicity. inhibit OCT2, peripheral peripheral selectively of severe would induces role (L-THP) (DRG) the accumulationL-tetrahydropalmatine ganglia high confirm the root however, to dorsal drug; aims platinum the anti-tumour in third-generation a OXA is of (OXA) Oxaliplatin Purpose and Background Abstract 2020 5, May 3 CN Hangzhou, 2 (86)-571-88208404 310058 CN CN Hangzhou, Zhejiang, Hangzhou, China 310058, 1 Ni Shixin hjagUniversity China P.R. Zhejiang Zhejiang Hangzhou, University, Zhejiang Hangzhou Road, Yuhangtang 866 Sciences Pharmaceutical of College University, Zhejiang 1 u Zhou Hui , 1 iigLi Liping , 2 uZeng Su , 1 efn Song Feifeng , 2 n ud Jiang Huidi and , 1 igagChen Mingyang , 1 3 1 ighnChen Yingchun , 1 igLi Ping , nvitro in nvivo in 1 , studies. studies. Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. ertxct scmlxaducer eue lttin,aclimmgeimmxue n oimion sodium peripheral a underlying its and mechanism limits mixture, the calcium-magnesium greatly Because a neurotoxicity patients. glutathione, peripheral of reduced unclear, life OXA-induced and of Therefore, ther- mg/m complex quality after is 1170 the years 2010). neurotoxicity affects even exceeds al., or & and OXA months et application (Argyriou several of clinical Zolota OXA for dose & lasts of cumulative which (Argyriou administration neurotoxicity, the apy peripheral first pre- chronic When neurotoxicity, the from peripheral after suffer of 2013). patients dysphasia, degrees al., and varying et paraesthesia, from Cavaletti suffer cramps, patients al., muscle 2016). 90% al., et as approximately et Trombatore sented that Najam & reported & carboplatin, is (Ruggiero neurotoxicity, (Bano and It application peripheral clinically cisplatin al., clinical especially ototoxicity as et neurotoxicity, its such no Grisold high-incidence limited drugs, the greatly & and almost platinum has severe for (Staff and of OXA-induced cancer used nephrotoxicity However, generation ovarian is mildest second 2013). and drugs, and the cancer antitumour first shows colorectal platinum the it as of to such Compared generation tumours, third 2017). solid the multiple to of belongs treatment which (OXA), Oxaliplatin dodecyl sodium SDS, medium; Introduction 1640 oxaliplatin; Institute OXA, resistance-associated Memorial error transporter; Park multidrug standard Roswell cation/carnitine SEM, MRP, RMPI, organic sulfate; saline; kidney; OCTN, buffer canine transporter; phosphate Madin–Darby cation PBS, organic HBSS, MDCK, serum; OCT, mul- extrusion; MATE, bovine protein; levo-tetrahydropalmatine; Foetal toxin L-THP, FBS, dehydrogenase; and medium; lactate Eagle LDH, tidrug solution; modified salt Dulbecco’s balanced DMEM, Hank’s ganglion; root dorsal DRG, L-THP. of use neurotox- clinical peripheral Abbreviations new OXA-induced a attenuate reveal to results candidate The efficient and 2. safe a find to us icity. help results The 1. OXA. of efficiency antitumor significance the Clinical impair OCT2. not inhibiting does by L-THP mainly 2. neurotoxicity peripheral OXA-induced attenuates L-THP 1. DRG. the in adds: OXA study of this accumulation What the in roles key play Transporters known: neurotoxicity. already is peripheral OXA- OXA-induced What attenuate attenuates to L-THP drug summary: DRG. candidate point potential the Bullet a in is uptake L-THP OXA OXA. of to efficacy contribute neurotoxicity OCT2, peripheral especially induced OCTN1, the and reduce OCT2 not cells). tumour- SW620 HT29 did in and Implications L-THP OXA (HT29 and of cells manner. efficacy Conclusion tumour antitumour dose-dependent in the a impair nor not in mice from did nude mice concentration-dependent and efflux bearing in tumours its a the DRG reduced affect in in and the not concentration neurotoxicity platinum OXA did in peripheral but concentration of OXA-induced cells platinum attenuated important accumulation DRG the most L-THP primary cellular Furthermore, the rat played and cells. OCT2 and MDCK-hMRP2 cytotoxicity OXA; MDCK-hOCTN1, of MDCK-hOCT2, the uptake in the reduced manner in involved L-THP were OCTN2 not role. but OCTN1 and OCT2 Results Key via niiigOT n CN u ihu marn h antitumour the impairing without but OCTN1 and OCT2 inhibiting 2 2 prxmtl 40-50% approximately , Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. Rswr olce ncl aksblne atslto HS) l prtoswr efre quickly performed the were and operations separated, All were (HBSS). canals operating solution spinal the salt The on balanced ethanol. placed Hank’s 75% were cold with g) in disinfected collected ˜250 were and (male, by DRGs anaesthesia rats described ether Sprague-Dawley methods after Healthy the table modification. to some according with isolated 2004), were cells cells DRG DRG maintained rat were cells primary All of penicillin/streptomycin. Preparation 1% grown and and air/CO FBS HT29 USA) humidified VA, 10% and Manassas, a with (ATCC; SW620 and supplemented in Collection laboratory medium penicillin/streptomycin. Culture our 1640 Type 1% in American GI and established the in FBS all (MDCK- from were 10% obtained cDNA cells) were with MRP2 (mock lines supplemented human vector cell medium containing blank and DMEM pcDNA3.1(+) 2017), in vector (MDCK- al., grown cDNA plasmid et OCTN1 Yang 2017), human & al., containing (Ma et pcDNA3.1(+) hOCTN2) (MDCK- Weng vector cDNA plasmid & OCT2 2017), (Li human al., hOCT2) containing et pcDNA3.1(+) Ma vector & (Bai plasmid hOCTN1) with L-THP transfected USA). stably China). MO, cells (Shanghai, MDCK Louis, Ltd. I Co., (St. (type Reagent Sigma-Aldrich collagen Aladdin culture from and from Cell obtained purchased C0130-1G) were were D109812) A10483-01) (No. (No. (No. from I No. OXA purchased Medium Collagenase were and tail, Neurobasal-A 8119040) USA). rat 2004470), medium (No. CA, from medium Eagle’s (Carlsbad, solution 1640) (No. modified (GI Corporation B-27 1640 Dulbecco’s Invitrogen 1816801), Institute 27250018), Gibco Memorial Park (No. (No. Roswell L-Glutamine and trypsin 1995169), THP5218), 10099141), No. No. (DMEM, (FBS, serum bovine Foetal reagents and Chemicals present the OCT2; effect. alleviate of subsequently antitumour Methods and activity its OCTNs the impairing and OCT2 inhibit without inhibit neurotoxicity strongly selectively peripheral would could L-THP OXA-induced not antitumour L-THP whether but its explore that OCTN1 to influencing carcinomas showed aims and without lost research colon study OCT2 neurotoxicity even previous 2015). of of or peripheral down-regulated Our al., inhibition OXA-induced tissues is and the et attenuate cells effect. malignant therefore, Li may tumour 2009), human 2014), most & MRP2 in G, al., (Myint and as OCT2 (Ciarimboli cells CTR1 of such et DRG modification expression in tumours, epigenetic from Blair the uptake and OXA to most & 2006), due of OXA in al., efflux (Larson mediate et expressed Varki the 2011) (CTR1) & in 2014; also (Holzer al., role 1 G, is et crucial Cavaletti transporter CTR1 Nakanishi a 2013; plays & Because ion MRP2 al., (Jong been copper while et (OCTN1) cells, has 2010), Ciarimboli 1 DRG It & transporter al., (SLC). (Sprowl cation et transporters (OCT2) carnitine/organic Zoumaro carrier 2 solute & transporter and cation Burger neurotoxicity. organic peripheral transporters OXA-induced ATP-binding that attenuate influx multiple reported to transporter-mediated strategy express the efficient al., of cells an et inhibitor & DRG Okabe probably selective (Johnson & is a OXA (Hall cells of of OXA DRG co-administration transport into of the transcellular OXA permeability Therefore, the of passive the in 2012). poor in roles al., accumulated the in crucial et be to platinum play Jun can Due of probably platinum concentration that 2000). transporters the indicated al., membrane which that et 2008), plasma, found McKeage the chemotherapy-induced was & in in It (Screnci that vulnerable than DRG symptoms 2015). particularly higher much sensory al., is was et primarily it DRG Canta the of barrier, & occurrence blood–brain (Carozzi al., the neurotoxicity the et by Postma peripheral explaining to & provided (Avan damage, critical protection (DRG) neurotoxic ganglion is the root to lacks it dorsal Entrena DRG spinal OXA, the Because & of of neurons 2015). Nieto use these sensory the however, clinical 2011; damages neurotoxicity; wide al., primarily neurotoxicity. OXA peripheral the peripheral et of OXA-induced Considering Pereira for alleviation & efficiency. strategies clinical new sufficient Descoeur the explore show 2010; for not evaluated al., recently did et interventions were A 2008) Galimberti al., & et (Scuteri blocker channel 2 nuao 5 /)a 37 at v/v) (5% incubator ° C. 3 ukye al. et Burkey Bre iggne al., et Hingtgen & (Burkey Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. vlaewehrLTPc-ramn 1 n 0m/g ol marteattmu ffiayo OXA of efficacy to antitumour groups the 4 impair into would allocated mg/kg) were 20 mice and nude (10 tumour-bearing co-treatment the L-THP activity, whether antitumour the evaluate the and sacrificed of co-treatment with were evaluation the after animals or For For recorded the without and analysis. administration, glucose 25). monitored for final 5% and were collected the weight of 22, were after body volumes DRGs 18, hours and equal 15, Twenty-four behaviour received mouse 11, administration. groups The 8, every L-THP-alone respectively. group, 4, or ( mg/kg), OXA-alone 1, vehicle (10 injected (days the group, L-THP intravenously weeks in vehicle was four Mice OXA female): of weremg/kg). randomly total half mice group. a were and the for L-THP-alone male feeding, groups, week and (half adaptive a each groups, one-week twice mice co-treatment mg/kg) with 16 L-THP mice with and L-THP, groups OXA of 6 effects into the neuroprotective into allocated cells the HT29 evaluate with To injected conditions. subcutaneously (SPF) were pathogen-free mice experiments. the 10 nude specific the Company x from the Technology under (1 week, Animal purchased maintained flanks one Laboratory were rear and for River Nude 8), Vital feeding 2012-0001) 2014-0001]. Beijing adaptive female, [Jing] (Zhe) from After 8; SCXK [SCXK obtained Sciences (male, were China; Medical male) weeks (Beijing, of old, 4 Academy weeks Zhejiang aged (5 All the mice suffering. g), of animal Center (22-25 Animal minimize mice Experimental to ICR made pathogen-free were Specific efforts CO all (22 by and temperature killed anaesthesia, controlled were hydrate mice a chloral at University the under Zhejiang cages with of performed in accordance Committee housed strict Use in and were Care ± out Mice Animal carried Institutional and Center. the double-blind for Medical were Guide the protocols of study rapidly. recommendations and 100 buffer procedures using incubation animal lysed the and All removing HBSS by ice-cold with terminated times treatment and three Animal MRP2) rinsed quickly transporter were efflux cells (for the trans- h Next, and 1 air/CO or mock humidified porters) a (for in ˜90% plates performed 40 pre- was to 12-well containing were experiment cultured in cells HBSS and DRG USA) of seeded 37 NY, primary mL were minor at Costar, and HBSS with cells (Corning mock, with cells) method The MDCK-hOCTN2, incubated DRG MDCK-hOCT2, reported (for MDCK-hOCTN1, previously plates 2017). 6-well a confluence. al., or to cells) et according transfected al., Yang evaluated porter et & was Li (Ma OXA & of (Tu modifications assay accumulation MTT cellular the The (LDH) by dehydrogenase evaluated lactate was study of viability uptake measurement 2016). cell Cellular the al., the for et and collected Song was kit, & medium assay Li cell the 2014; LDH a of the to 40 part using with adjusted a treated activity h, were medium, cells 48 fresh the another h, in for 24 resuspended for logarithmic cultured were were the cells cells DRG The in collected. cells 4.0 and of mock density digested and were 96-well phase MDCK-hMRP2 in growth MDCK-hOCT2, well MDCK-hOCTN2, per cells MDCK-hOCTN1, 7500-8000 or plates 37 study at tissue-culture Cytotoxicity maintained 6-well were in cells well The 50 per plates. containing cells tissue-culture medium 37 28,000–30,000 a min, in at (90 dispersed plated IV were collagenase cells mg/mL DRG 1.25 the with digestion After ice. on 0)wt 2hlgtdr yl n rvddwt reacs ofo n ae.Alsreiswere surgeries All water. and food to access free with provided and cycle light/dark 12-h a with 10%) × 10 7 4 m,adsee n9-elcl ltsa . L/el fe h bv el n primary and cells above the After /well. mL 0.2 at plates cell 96-well in seeded and /mL, el/ie.Nd iewt uor faeaevlm 06 mm 50-60 = volume average of tumours with mice Nude cells/side). iv 2 shxain nml eernoie o treatment. for randomised were Animals asphyxiation. netdwt %guoeslto otiigLTP(-0m/g n X (8 OXA and mg/kg) (5-20 L-THP containing solution glucose 5% with injected μ ° o i,wieMC-MP el eeicbtdfr3 i.Nx,0.5 Next, min. 30 for incubated were cells MDCK-hMRP2 while min, 5 for C X iho ihu -H a de oiiit ellrutk.The uptake. cellular initiate to added was L-THP without or with OXA M 2 nuao 5 /)a 37 at v/v) (5% incubator 4 ° ne %C2atmosphere. CO2 5% a under C μ -uo2-exuiie esr ern were neurons Sensory 5-fluro-2’-deoxyuridine. M μ X nteasneo rsneo L-THP of presence or absence the in OXA M ° )ad005 rpi 9 i,37 min, (90 trypsin 0.025% and C) ° o i frutk trans- uptake (for min 5 for C ± 0.1 ° )adhmdt (50 humidity and C) μ f00%SDS. 0.01% of L 3 eeslce for selected were iv, ° C), 8 Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. anr ihices ncl iblt n euto fLHlaaei h elmdu;priual,i the in particularly, medium; cell the in leakage and LDH MDCK-hOCTN1 of the reduction In and viability respectively. cell folds, in LDH 4.0 increase the and with 1.7, while manner, 1.1, respectively, MDCK-hOCT2 1-100 to cells, as increased cells, MDCK-hOCT2 were well MDCK-hOCT2 and inhibitory medium as MDCK-hOCTN1, the the cells, in mock, studied MDCK-hOCTN1 activities the we and in OCT2, 40 mock treatment) of above, (vehicle in inhibitor described toxicity we strong OXA-induced As a on cells. was OCT2- L-THP L-THP in of that OXA effect demonstrated of study previous accumulation Our the of reduced uptake cells and the OCTN1-transfected toxicity to and contribute OXA-induced ( OCTN2 cells attenuated role. not mock crucial but L-THP the most OCTN1 in the and that plays OCT2 folds OCT2 that 2.0 and imply OXA, and results 5.4 above were The cells respectively. MDCK-hOCTN1 and MDCK-hOCT2 the okcls(LD cells mock oiiyi rnpre-rnfce DKcls u eut eeldta h re fsniiiyt OXA OXA-induced to compared sensitivity we of OXA, order of the uptake that the revealed in results OCTN2 (LD Our and MDCK-hOCT2 cells. OCTN1, was MDCK OCT2, transporter-transfected of in cells toxicity roles MDCK-hOCT2 in the accumulation confirm and To toxicity highest the exhibited OXA (GraphPad 5.0 Prism Results GraphPad USA). using CA, performed Diego, were San analyses Inc., data Software All significant. statistically considered a sdt nls ieecsbtentogop,adoewyaayi fvrac AOA followed (ANOVA) variance of Student’s analysis two-tailed one-way Unpaired and each. groups, samples two Dunnett’s six between or or differences Bonferroni’s five by analyse with to (LD experiments used dose independent was lethal two median least viability The at means cell the plotting blinded. by as were fitting analysis curve data sigmoid and experiments All tissues. for analysis weight Statistical the was to solution relative the or after (0.22 cells 300X) filter for Nexion film PerkinElmer a (ICP-MS, ddH using 200 spectrometry with filtered Next, mass mL plasma 4.0 . to coupled 80 adjusted inductively at was solution h the 2 of for of incubated frequency was the mixture min, 26). 200 1 the added and for ICP-MS We of adapt by 23, temperature to concentration min. 19, The mice OXA 2 the for 16, of floor. allowing recorded Determination Biological 12, cold-plate After was Zhenghua the degC. mice 9, the 4.0-4.2 on (Anhui by 5, at placed cold-plate licking constant were 2, hind-paw ZH-6C kept mice (days was the The plate administration using cold Ltd.). after assessed Co., was h Equipment threshold 24 Instrument nociceptive performed thermal were The tests behavioural All body were sizes the animals tumour. tumour and the the The tests 13), of administration, Behavioural analysis. side and final short for = were 11, the stored size the 9, is Drugs tumour and after b 7, formula: weighted, and following hours collected, 5, groups. the Twenty-four were to 3, co-treatment tumours according 1, calculated L-THP the monitored. (days and and were weeks sacrificed, OXA size two were tumour and for and days group, two weight OXA-alone every group, intravenously vehicle administered mg/kg): (8 ± μ 50 fHNO of L tnaderro h en(E) o the For (SEM). mean the of error standard 100 ¿ μ )( M) 50 3 3.3 : n 400 and μ othoc post fLTPatnae X-nue oiiyi concentration-dependent a in toxicity OXA-induced attenuated L-THP of M iue1a Figure μ ± ) h X otn a eemndrltv otettlpoencontent protein total the to relative determined was content OXA The m). μ 0.7 fOArdcdtecl iblt o7% 0,ad1%o h control the of 12% and 50%, 76%, to viability cell the reduced OXA of M μ fH of L μ etwsue o utpegopcmaios A comparisons. multiple-group for used was test )¡MC-OT1(LD MDCK-hOCTN1 ¡ M) .W ute hwdta h ellracmlto fOAin OXA of accumulation cellular the that showed further We ). 2 O 2 versus notelst ftecletdcls Rs rtmus the tumours; or DRGs, cells, collected the of lysate the into 2 .Fnly h ocnrtoso X eedtrie by determined were OXA of concentrations the Finally, 0. μ f2%amnaslto a de,adtevolume the and added, was solution ammonia 25% of L 5 h o1 X ocnrtos aaaepresented are Data concentrations. OXA log10 the a * nvitro in b 2 /2 50 32.5 : hr steln ieo h tumour the of side long the is a where , aa ahpitwsotie from obtained was point each data, ± 1.1 μ 50 )¡MC-OT2[?] MDCK-hOCTN2 ¡ M) a bandthrough obtained was ) P -value iue1b Figure < .5was 0.05 t test ), Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. oepoewehrLTPmgtipi h niuorecec fOA h yooiiyadantitumour and cytotoxicity the OXA, of efficiency tumour-bearing antitumour in the impair nor might cells L-THP tumour whether in explore To OXA of rather efficacy cells antitumour L-THP mice DRG the nude Therefore, impair in not OXA did excluded. of L-THP uptake be the could inhibiting L-THP abirritation. by by of neurotoxicity than L- abirritation peripheral while ( the manner mg/kg), OXA-induced mg/kg) (8 that ameliorated (5-20 OXA indicating dose-dependent with a sensitivity, treated in stimulation DRG mice the in in ng/mg platinum 6c 2.35 of concentration reached the DRG reduced the THP was in dose-dependently iv), mg/kg) platinum ( mg/kg, (5˜20 sensitivity L- of L-THP (8 cold-stimulation OXA concentration with OXA-induced mice. with female in treated or or increase male mice the without vehicle-treated the attenuated the in mice in licking that hind-paw OXA-treated folds 3 of in approximately frequency in the DRG as shown in expressed As stimulation, accumulation neurotoxicity platinum co-treatment. OXA-induced detected THP attenuate and could test L-THP behavioural whether confirm DRG further To in accumulation thereby platinum can reduced mice and ( and OXA of neurotoxicity group of OXA-induced OXA-alone accumulation ameliorated the cellular cells. L-THP in the DRG 100 that primary decrease in can at of toxicity L-THP L-THP 32% OXA-induced that example, to ameliorate suggest for cells results DRG manner; our concentration-dependent summary, primary (1-100 a in L-THP in test, accumulation viability platinum cells cell DRG the primary of result in the with agreement In L-THP cells. with DRG co-treatment in OXA while and of control), toxicity accumulation the of OXA 52% reduce approximately μ could (to strongly L-THP cells L-THP 40 DRG that and (1-100 that primary respectively, revealed deduced of OXA, results we viability of Our efflux cell DRG. MRP2, and in not toxicity uptake OXA-induced the but attenuate in activity roles OCT2 vital DRG play inhibited primary MRP2 and in OCT2 toxicity Since OXA-induced attenuated and accumulation cells OXA MRP2. reduced MRP2. of L-THP inhibiting inhibition by by efflux efflux evaluated OXA OXA we decrease reduce 1-100 cells, not would and DRG L-THP cells, from whether mock OXA elucidate the of to efflux in MRP2 the shown on mediating As L-THP transporter of important effect MDCK-hMRP2 most the in the OXA OXA is OCTN1-mediated of MRP2 accumulation Since and cellular OCT2- inhibit inhibit not significantly did L-THP can toxicity. OXA-induced L-THP hOCT2 attenuate of that subsequently inhibition and implies stronger ( uptake considerably cells which a mock co- exhibiting hOCTN1, the uptake, whereas in than OXA respectively, OCTN1-mediated that cells, and folds mock OCT2- 2.8 in inhibits and that 1.1 folds to of 5.4 cells half and (40 to 2.0 100 OXA reduced with approximately of incubation was was accumulation cells LDH cellular MDCK-hOCT2 the the and and hand, 12%) other (vs the 64% On ( to group increased OXA was the viability in that cell the cells, MCDK-OCT2 fLTPicesdtecl iblt o7%o h oto,wihidctsta -H a euethe reduce may L-THP that indicates which control, the of 75% to viability cell the increased L-THP of M .TeLTPaoegopadtevhceaoegopddntso infiatdffrnei cold- in difference significant a show not did group vehicle-alone the and group L-THP-alone The ). μ )icesdtecl iblt nacnetaindpnetmne ( manner concentration-dependent a in viability cell the increased M) iue4 Figure μ -H eue h X cuuaini h DKhCN n MDCK-hOCT2 and MDCK-hOCTN1 the in accumulation OXA the reduced L-THP M h cuuaino X nMC-R2clswsapoiaeyhl fta in that of half approximately was cells MDCK-MRP2 in OXA of accumulation the , iue2a-2f Figure μ -H i o nraeteacmlto,wihidctsta -H does L-THP that indicates which accumulation, the increase not did L-THP M iue6 Figure ). 4hatrtefia diitain h estvt ocold to sensitivity the administration, final the after h 24 , iue3 Figure 6 μ .Terslsidct htLTPsignificantly L-THP that indicate results The ). ,3 i nuain nteMDCK-hOCTN1 the in incubation) min 30 M, μ iue6 n 6b and 6a Figure )rdcdteacmlto fOXA of accumulation the reduced M) μ fOAsgicnl eue the reduced significantly OXA of M iue5a) Figure nvivo in .Acrigy the Accordingly, ). iue5b) Figure epromda performed we , o ntne 10 instance, for ; μ eue the reduced M Figure In . Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. u rsn eut hwdta oicbto ihLTPddntdces h eltxct fOAin OXA of al., toxicity et cell Qin The the OXA). & decrease of Chen not indication 2019; clinical did al., main down-regulated expected, L-THP et As (the extremely with Yu OXA. lines is of & co-incubation cell effect it (Zhu antitumour that cancer the DRG, modification showed colorectal impair the epigenetic not results would in to present OCT2 uptake due of our OXA inhibition tumours selective efflux. in most Therefore, its role than in 2019). rather crucial lost DRG most even the the or in Since plays uptake DRG 2016). OXA OCT2 the study. al., inhibits in present Although et mainly accumulation the L-THP Song OXA in decreased that & considered significantly speculate (Li not L-THP we MATE1 was that ATP7A/7B , of result model, function the cell the ATP7A/7B-transfected on affect an Based have not OXA tissues L-THP of not did tumour that the one did L-THP and indicating that we as 100 DRG that cells, showed the and revealed mock reported result in and study cells, was 2011; Our accumulation cells previous mock MRP2 OXA al., MDCK-MRP2 in cells. and influence 2019), in et that not DRG OXA DRG, Koepsell al., than of does from the & et lower accumulation OXA in (Nies much the Nardella of MATE1 efflux influence was & obviously and efflux OXA cells Lasorsa 2019), the MDCK-MRP2 in 2018; al., in involved in al., et accumulation transporters are Biswas et important 2019) & Yin most al., is Myint & the et it 2015; Li Hirota therefore, al., 2010; & et CTR1; Fujita al., Li inhibit et & not (Myint in OXA. Liu did that MRP2 of & L-THP with efficiency (Ip that antitumour compared ATP7A/7B the when implies Additionally, impair altered, which in not obviously alone, tumour did expressed not OXA L-THP was the also with OXA that in is reasonable and treated platinum OXA-induced CTR1 L-THP of of mice inhibition that with concentration nude the the co-treated the Considering the that for mice that the DRG. target indicates nude Liu demonstrated good which in in the & a result cells, essential not CTR1 of (Ip study DRG is be histidine Our of it in not neurotoxicity. carcinomas, copper uptake role colon may peripheral OXA CTR1, the as uptake on of such study tissues, OXA influence inhibitor malignant significant not in the human a did al., CTR1 exhibit et we of not Burger role model, did by 2013), cell reported al., CTR1-overexpression as et However, a OXA. have cells of not MDCK-hOCTN1 uptake in did cells). that mock LD we than the the Because cells of because MDCK-hOCT2 that OXA in folds that of higher 2.0 not uptake much vs but the was (5.4 OCT2 OXA in estimated and of role first OCTN1 accumulation (3.4 important we of cellular cells more OXA, substrate and MDCK-hOCT2 a a of MDCK-hOCTN2, played in is uptake OCT2 OXA MDCK-hOCTN1, OXA Furthermore, the uptake mock, that in OCTN2. demonstrated the in transporters data of accumulation to Our individual contribute OXA of CTR1 and cells. role toxicity MDCK-hOCT2 and antitumour the cell OCTN1, the compare OXA-induced OCT2, impair further the that not to shown did OXA, have it of papers therefore, published DRG, tumours; several the in Although in OXA OXA that of L-THP of demonstrated accumulation that OXA. accumulation results the of demonstrated the the our efficiency reduce study in Finally, reduce further not roles MRP2, neurotoxicity. The did vital OXA- in not peripheral OXA. play L-THP OXA however, but OXA-induced of OCT2, of attenuate activities efflux cancer; especially subsequently use OCTN1 the colorectal OCTN1, and and the in involved and as OCT2 limits is OCT2 inhibit such greatly MRP2 that could and tumours and confirmed patients OXA, solid study of of present of uptake quality The treatment life the the clinic. the reduces for neurotoxicity drug peripheral effective induced an the is in no OXA as showed tumours well in as groups. OXA group, results co-administration of dose) the L-THP concentration Discussion or mice the with single OXA-alone Meanwhile, agreement nude mg/kg, the groups. In (8 tumour-bearing between mg/kg) OXA-alone difference HT29 cells. 20 significant the SW620 and in in (10 or as co-treated inhibited HT29 well L-THP markedly the as was in growth cells, OXA tumour of SW620 in cytotoxicity and shown the HT29 As reduce L-THP. in without evaluated or were with OXA of effect μ )wsmc oe hnta nMC-OT1(25.8 MDCK-hOCTN1 in that than lower much was M) iue 7a Figures and 7 7b oicbto ihLTP(1-100 L-THP with co-incubation , nvivo in via niiigMP.Mroe,our Moreover, MRP2. inhibiting eut ute eeldthat revealed further results μ nvitro in -H i not did L-THP M μ nvitro in μ ) n the and M), and )ddnot did M) nvivo in the , 50 of Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. opud ncide ihcne:txct n lnclmngmn.At-acrDus[] Anticancer Platinum [J]. (2013). Drugs R. Anti-Cancer Riccardi management. . . clinical . and M, toxicity 1007-1019. Scalzone P, cancer: 24: Ferrara Drugs, with R, children Arena in S, Triarico compounds A G, data. Trombatore C. Neuropathy: analyzed A, Peripheral C. 772-781. Z. Ruggiero Chemotherapy-Induced Y. 81(6): S. C., (2017). Neurol, Z., Y. AJ. Ann H. M. Windebank [J]. and L. W, Review C. P. Grisold Current C. S., A, Y. F. Grisold paper. C. F. NP, the Y. Staff L., P. wrote M. L. J. L., P. Y., D. L. H. D. Y., and Y. References C. D. research. Y. Y. the M. designed research. Y., J. the D. D. performed Y. H. N. and X. L. S. OXA-induced P. and L. attenuate Y., to D. candidate Y. potential a likely the Contributions is in Authorship L-THP OCTN2 neuro- studies, not peripheral clinical but Considering OXA-induced OXA. in prevent neurotoxicity. OCTN1 of could effect L-THP peripheral and antitumour L-THP of the OCT2 impairing that the safety without of demonstrated in the OCTN1, role and and role OCT2 DRG crucial inhibiting important the through the an toxicity in confirmed play OXA Apart study to of present uptake determined we the membrane. been permeability, summary, consideration. also mitochondrial passive deserves In has shown). the which high family 2017), not its across (Taylor, SLC25A (data to system transport DRGs the carrier due OXA family, rat mitochondrial membrane SLC22A of inhibited cell the mitochondria probably the from the L-THP across in analysis easily that expressed blotting localized passes OXA western speculated were be how our L-THP OCTN1 to identify and that and reported 2006), to & Considering been OCT2 Tein, meaningful Trecarichi has & that and OCTN1 (Lamhonwah 2019; showed interesting permeability. membrane also al., passive mitochondrial is et poor mammalian it its the Xiang 2018), of in & al., because (Cheng mitochondria et tumour the Shimoyama dysfunction the enters & and mitochondrial Toyama tumours induce 2019; the can Flatters, in efficiency OXA groups. concentration antitumour co-treatment that platinum the L-THP Considering impair the experiment was and the not comparing group group terminate did from OXA-alone to L-THP same results the had that we between the conclude the that size can on fast from mice we based Fortunately, so nude mice grew OXA, tumour-bearing weeks. group two of in female vehicle of OXA the and end of difference; in the effect male tumours gender antitumour at behavioural the of no the The however, evaluate weeks; showed DRGs sample. to 3 neurotoxicity one aimed for the detection as peripheral had the in we pooled OXA-induced Originally, of were concentration that requirements combined. group platinum the showed same meet the 5b) the To of and therefore, test. mice 5a behavioural two the (Figure from allocated in DRGs were test female) the mice half platinum, 16 neurotoxicity. and of difference, of peripheral male limit gender OXA-induced effect (half exhibits inhibitory of group neurotoxicity the inhibition one peripheral that in the proved OXA-induced in directly whether factor L-THP study The important by To the effect. mice sedative/hypnotic be the may its of to OCT2 DRG cold-stimulation irrelevant on the of was in L-THP reduction L-THP OXA the co-administered of that was reduction and implied that significant observation group group control above the vehicle The in the alone. As between sensitivity hypnotic change L-THP or alone. significant sedative administered L-THP exhibit no group with not showed Tothe evaluation did treated mice behaviour paws. was neurotoxicity the further hind that administration, peripheral and group lapping L-THP OXA-induced effects, a the of 2017). after included frequency hours al., we twenty-four the et L-THP, observed, in Du we of (Liu increase & effect years Du an sedative/hypnotic 40 2016; by the than al., exclude indicated more et as for Yue sensitivity, properties & sedative/hypnotic cold-stimulation Gong with antitumour increases 2019; analgesic the al., an et as impair Wang China and & in OXA used been of has accumulation L-THP tumour the reduce OXA. not of efficacy did L-THP of co-administration 8 Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. nu rnpre nnra n ainn ua ise J.JHsohmCtce,5:1041-1049. copper 54: human Cytochem, the Histochem of J Expression [J]. tissues (2006). human SB. malignant Howell P, and Naredi normal MA, in Gibson, 1 7. QT, transporter 1(10): Le influx organic One, NM, Mediated Varki PLoS by (MRP2) AK, [J]. 2 Vesicles Holzer mediated Protein Membrane Resistance-Associated transport in Multidrug 537-547. cells Platinum (2015). OXA 338(2): OXA-Derived M. 293 Ther, of McKeage Exp kidney J, Transport Paxton Pharmacol (2011). embryonic Y, J Li MJ. human [J]. K, Myint overexpressing neurons McKeage ganglion in I, root OCTN2 Tamai dorsal the rat in and JJ, and 1 OCTN1 Liu transporter transporters copper T, mammalian 324-330. cation/carnitine Nakanishi the 75(2): of NN, Pharmacol, role Mol The Jong [J]. (2009). (2010). drugs SB. platinum-based EA. Howell of Wiemer R, accumulation Safaei cellular BG, . . Blair . (hSLC22A2) CA, RH, hOCT2 Larson 898-908. transporter Mathijssen 159(4): cation EM, organic Pharmacol, human Berns J the Br J, by [J]. compounds Helleman platinum AW, of we Boersma transport what DA, Differential and Zoumaro 66-76. need H, 19(2): we Burger Syst, what USA, Nerv (CIPN): Sci Peripher neurotoxicity Acad J peripheral [J]. Natl OXA know Chemotherapy-induced Proc (2013). (2014). [J]. A. OCT2 G. Sparreboom Cavaletti transporter . . cation . organic G, Du the AA, 11199-11204. on Gibson 110: dependent H, is Giovinazzo of neurotoxicity CS, Pharmacology Lancaster induced in the 962-986. G, of 12: accumulation Ciarimboli Targets, Determinants JA, cellular as Drug Sprowl Transporters Cancer of Membrane Current role (2012). [J]. JM. The Drugs 495-535. Mark Anticancer (2008). and 48: Platinum Toxicol, MM. L Pharmacol Jun Gottesman Rev JL, Annu XJ, Johnson [J]. Liang chemotherapy DW, platinum-based to Shen Br. sensitivity M, [J]. determining Okabe drugs platinum MD, of betweenHall series Relationships a (2000). 966-972. of BC. 82: toxicity Baguley Cancer, nerve BD, J. Palmer peripheral TW, and Hambley accumulation P, reactivity, Galettis hydrophobicity, MJ, McKeage D, know 90-107. we Screnci 596: do Lett, What neuropathy: Neurosci Platinum-induced peripheral [J]. (2015). Chemotherapy-induced 411-432. mechanisms? (2015). GJ. 20(4): about A. Peters Oncologist, Chiorazzi E, [J]. A, Giovannetti Canta future VA, G, and Carozzi Cavaletti 520-531. present, the A, 137(3): past, inhibits Pain, Avan strategies: Tetrodotoxin preventive [J]. C, and (2008). mice Ceresa neurotoxicity in JM. TJ, paclitaxel Postma Baeyens by A, JM, induced Med, Avan Vela pain Mol neuropathic ED, EMBO of Pozo OXA-induced [J]. expression Cend´an CM, (2011). and nociceptors JM, E. development in Bourinet Entrena expression FR, channel . . Nieto . ion V, of Maffre remodelling B, 266-278. to protects Ling 3(5): due A, NGF Francois is 486(3): A, hypersensitivity (2010). Lett, Pizzoccaro cold Neurosci G. V, [J]. Tredici Pereira ERK1/2 J, G, and Cavaletti Descoeur JNK/Sapk E, modulating Donzelli by OXA S, from Pasini 141-145. neurons M, ganglion Ravasi associated root A, neuropathy 435–446. dorsal peripheral Galimberti 15: Toxic BUON, A, J (2010). Scuteri [J]. HP. agents Kalofonos chemotherapeutic 3570-3577. O, pe- used OXA-induced Kyriakopoulou commonly 119: with Cancer, of V, (2013). [J]. development H. Zolota study Kalofonos the AA, multicenter in Argyriou prospective . . role a . pivotal from J, results Bruna a C, neurotoxicity: play Briani ripheral polymorphisms AA, channel Genazzani 1637-1641. A, sodium Reactions Voltage-gated Hypersensitivity Antonacopoulou 17(4): Induced G, Prev, Cavaletti OXA Cancer of AA, J Features Argyriou Pac Clinical Asian (2016). [J]. A. Approaches Mateen Therapeutic F, Qazi and R, Najam N, Bano 9 Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. n yntceet flttayrplaiei os erpti anmdl[] Psychopharmacology [J]. analgesic model mediate miR-630 pain receptors and D2 neuropathic miR-489-3p and mouse 1008-1020. D1 of a 3169-3182. Dopamine 9(5): Upregulation in B, 236(11): (2019). l-tetrahydropalmatine (2019). Sin ZL. (Berl), of L. Huang Pharm Yu effects WM, Acta Qu hypnotic [J]. JC, . and . OCT2 Zhou . targeting TX, H, Wang by Wang YY, carcinoma K, Liu cell Zeng renal S, repression in Ye transcriptional uptake J, OCT2 791-803. OXA of Lei inhibits Regulation 14(8): Z, Epigenetics, (2019). Qin [J]. S. L, Zeng carcinoma Chen X, cell Zheng renal H, to in Hu contributing acetylation L, transporters Chen histone drug Z, by Qin of L, Identification 373-385. Yu 105-167. Q, (2019). 148(3): Zhu I. Neurochem, (201): Ieiri J Pharmacol, M, [J]. Exp vitro Baba neuropathy Handb in R, peripheral [J]. MATEs), Sakiyama OXA-induced therapy a (OCTs, T, drug as transporters Hirota in MRP2 cation S, of importance Fujita Organic Identification the (2011). for (2019). 2245 M. evidence M. 9(1): Schwab vivo McKeage Rep, K, in Sci Damme and . [J]. . H, . cancer Koepsell gastrointestinal J, AT, in Liu Nies response S, and Jamieson accumulation N, OXA Jong limiting 7. Y, factor 1(10): Li targetable One, R, Mediated Biswas PLoS (MRP2) K, [J]. 2 Vesicles Myint Protein Membrane Resistance-Associated Am in Multidrug J Platinum Mech- (2015). [J]. OXA-Derived M. (2019). of McKeage Drugs F. J, Transport Anticancer Paxton Arnesano Platinum Y, Li by . K, Trafficking . Myint . Copper R, 12109-12120. of Caliandro Inhibition 141(30): R, Caliandro for Soc, biomarkers V, Chem Basis Novel 183-191. Mirabelli ATP7B: Structural 70(3): A, and and Life, ATP7A Rosato anistic IUBMB MI, transporters [J]. Nardella efflux in therapy A, Copper CTR1 for Lasorsa targets and (2008). and ATP7B XP. resistance ATP7A, Li drug ZQ, of 53. platinum Liu expression for 13(6): JY, Differential Pain, Yin Mol YQ, (2010). [J]. Li tissue MJ. ganglion McKeage root JF, dorsal Mercer rat JJ, adult Liu Pharmacol Exp V, Clin Ip [J]. 1-mediated vivo transporter in copper and on 371-378. histidine vitro copper in 40(6): of neurotoxicity Physiol, effects oxaliplatin-induced of and 189-202. Evaluation platinum MATE1 dorsal-root 99: (2013). of and MJ. the accumulation Med, McKeage from OCT2 Mol JJ, neurons of Liu Methods sensory V, Role [J]. of Ip rats culture (2016). 2543-2554. adult and HD. or 173(16): Isolation Jiang embryonic Pharmacol, (2004). of J MR. . ganglia Vasko Br . . CM, [J]. Transporter1 HM, Hingtgen chloride Cation TH, Lei nitidine Organic Burkey K, of of Involvement Wang toxicity X, and (2014). Yang H. disposition 34-42. YY, Jiang renal Weng in 322: Toxicology, . FF, . [J]. . Song Toxicity LP, S, Induced Sun Li Retrorsine Z, in Chen dorsal-root 3A4 Z, 189-202. the P450 Ma from Cytochrome 99: H, neurons and Med, Lei sensory L, Mol Li of Methods Transporters M, ABC culture [J]. Tu and rats and SLC adult Isolation Multiple or 269-278. transporters (2004). (2017). embryonic 45(3): cation MR. H. of Vasko Dispos, Jiang organic ganglia CM, Metab Multiple . Drug Hingtgen . . [J]. T, S, Entecavir (2017). Burkey Zeng of H. C, Transfer 526-534. Zheng Placental Jiang M, 38(9): the H, Bai Dispos, to T, Zhou Contribute Drug Jiang mediate Biopharm H, X, transporters Yang [J]. Lei drug Z, sulpiride Ma Multiple M, of (2017). transport Bai H. renal W, Jiang the Wang to Y, . contribute 3873-3884. . . Weng 91(12): X, Toxicol, L, Yang Arch Li Y, [J]. Weng sulpiride C, of Zheng transport D, placental Sun the Z, Ma 34(1): M, Res, Bai Anticancer [J]. side-effects cisplatin of mediators as 547-550. transporters Membrane (2014). G. Ciarimboli 10 Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. nteasneo rsneo X o 8h o h ellracmlto td,teclswr incubated were cells the study, accumulation cellular cells, the mock for the h; with 48 Compared for accumulation. incubated OXA OXA were of cells (40 presence the OXA study, with or cytotoxicity absence the For the cells. in MDCK hOCTN2-transfected hOCT2-, hOCTN1-, 1. Figure 27(9): Biol, Cell Trends [J]. System Carrier Mitochondrial the of to Properties transporter, 633-644. Functional cation/carnitine 1315-1325. (2017). organic 345(4): EB. an Commun, Taylor OCTN1, Res Biophys of Biochem localization [J]. Novel mitochondria (2006). mammalian I. Tein AM, Lamhonwah ACS 1566-1571. [J]. Mitochondria- Mice 9(7): a in of Neuropathy Neurosci, Peripheral Effect Chem Chemotherapy-Induced Protective of Development (2018). the M. Shimoyama against PW, Peptide Targeted Schiller HH, Szeto 83-126. N, chemotherapy-induced Shimoyama 145: of S, Toyama pathogenesis Neurobiol, via the Rev 3140-3149. neurotoxicity in Int dysfunction 11(5): OXA-induced [J]. Mitochondrial neuropathy 2019, ameliorates peripheral (2019). Res. IIA Transl SJL. Tanshinone J Flatters A, inhibits Am (2019). Trecarichi [J]. Levo-tetrahydropalmatine K. promotion Xu (2017). autophagy S, and KM. and Wang Yun protection ERK 367-373. mitochondrial W, of 15(317): Xiang . expression . Res, . W, the Brain H, Cheng regulating Behav Su by [J]. Y, natural, rats preference Feng in a place phosphorylation C, Levo-tetrahydropalmatine, conditioned CREB Holscher ketamine-induced (2016). J, of C. Cao acquisition Li the L, Du 67-72. . . Y, . 144: Du D, Behav, Wang Biochem Pharmacol Y, methamphetamine- and [J]. Gan self-administration reinstatement J, methamphetamine induced inhibits Xing antagonist, B, receptor dopamine Ma mixed K, Yue X, Gong ocnrto-eedn yooiiy()adclua cuuain()o X nmc and mock in OXA of (b) accumulation cellular and (a) cytotoxicity Concentration-dependent μ )fr5mn aaaeepesda h mean the as expressed are Data min. 5 for M) ## 11 P < 0.01, ± ### E,n=6frcl oiiyadn=5for 5 = n and toxicity cell for 6 = n SEM, P < 0.001. Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. rpeec fLTPfr5mn aaaeepesda h mean the as expressed are Data (40 OXA min. with 5 incubated for were cells, L-THP cells of the study, presence accumulation or the In cells. MDCK-hOCT2 3. Figure mean the as expressed are Data h. the 48 study, toxicity for the group, L-THP vehicle In and/or the cells. with OXA Compared 0.01and (f) activities without 5. MDCK-hOCT2 LDH = or and n cells. with (e), SEM, (c) MDCK-hOCTN1 incubated MDCK-hOCT2 (d), were and mock MDCK-hOCTN1(b), cells for (a), medium mock the of in viability Cell cells. hOCT2 2. Figure ### *** P P < h ffc fLTPo X-nue yooiiyi ok DKhCN,adMDCK- and MDCK-hOCTN1, mock, in cytotoxicity OXA-induced on L-THP of effect The h ffc fLTPo h ellracmlto fOAi ok DKhCN,and MDCK-hOCTN1, mock, in OXA of accumulation cellular the on L-THP of effect The < .0;cmae ihteOAaoegroup, OXA-alone the with compared 0.001; 0.001. ### P 12 < .0;cmae ihteOAaoegroup, OXA-alone the with compared 0.001; * P < .1and 0.01 ± E,n=5 oprdwt h mock the with Compared 5. = n SEM, ** P < 0.01, *** P μ )i h absence the in M) < 0.001. ** P < ± Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. a,fml ie()adOAacmlto nteDGi edrmthdmc c.Dt r xrse as expressed are Data (c). mice matched gender in * DRG the group, in accumulation OXA mean and the (b) mice female (a), 6. Figure group, vehicle the with Compared study. accumulation platinum mean group, the the OXA-alone as for the expressed 5 with are = compared Data study. n accumulation and (40 the OXA study in with min 5 or for without and incubated were Cells cells. 5. Figure (40 mean OXA the with as incubated were cells The 4. Figure P ± < h ffc fLTPo yooiiy()adclua cuuain()o X npiayDRG primary in OXA of (b) accumulation cellular and (a) cytotoxicity on L-THP of effect The h ffc fLTPo h ellracmlto fOAi okadMC-MP cells. MDCK-hMRP2 and mock in OXA of accumulation cellular the on L-THP of effect The h ffc fLTPo X-nue rqec flpighn asicessi aemice male in increases paws hind lapping of frequency OXA-induced on L-THP of effect The E,n=8 oprdwt h eil group, vehicle the with Compared 8. = n SEM, ± 0.05, E,n=5 oprdwt h okgroup, mock the with compared 5; = n SEM, ** P < .1 and 0.01, *** P * P < μ )i h bec rpeec fLTP h aaaeexpressed are data The L-THP. of presence or absence the in M) < 0.001. 0.05, ** P 13 μ < )ado -H o 8hi h yooiiystudy cytotoxicity the in h 48 for L-THP and/or M) .1 and 0.01, ### ### P P *** < < P .0;cmae ihteOAalone OXA the with compared 0.001; 0.001. < 0.001. ± E,n=6frtecytotoxicity the for 6 = n SEM, ### P < 0.001; Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. nattmu ffiiny()o X cuuaini uor d nH2 uorbaignd ie Data mice. nude group, tumour-bearing HT29 vehicle in the mean (d) with tumours the in as accumulation expressed OXA are or (c) efficiency antitumour on 7. Figure h ffc fLTPo h niuo ffiinyo X nH2 a n W2 el b and (b) cells SW620 and (a) HT29 in OXA of efficiency antitumor the on L-THP of effect The ### ± P E,n=6frtesuyi el n o hti uemc.Compared mice. nude in that for 8 = n and cells in study the for 6 = n SEM, < 0.001. 14 Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. 15 Posted on Authorea 26 Feb 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158274612.26366163 — This a preprint and has not been peer reviewed. Data may be preliminary. 16