Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. fflxtasotr nnnsalcl ugcne patients Liu cancer Lu lung cell and non-small CYP1A1 in involving transporters gefitinib efflux and ningetinib of interaction Drug htti td adds: study this What proposed clinically was gefitinib and ningetinib with therapy treatment. drug NSCLC combination the a for efficacy, the subject: improve this To about known already profile safety patients. is long-term NSCLC What not The in was concerned ningetinib ningetinib. drug be of parent should concentration the M1 gefitinib tissue of on with target pharmacokinetics combined drug the the ningetinib parent vivo, increase of the in its may efficacy of M1 reduced CYP1A1 and of effect and the yield M1 inhibitory high of of metabolic the Inhibition the low formation and Consequently, the the influenced. affinity to inhibited tissue due gefitinib transporters. low but co-administered, efflux When its exposure, these efflux plasma to Implications: the of attributed and of inhibitor Conclusion was substrate 0.095 patients an the efflux. in as was canalicular of identified M1 ningetinib value was the of while M1 Ki study exposure MRP2, mice. with plasma to in and 75% CYP1A1 used BCRP drug by formation. of were P-gp, D6-M1 M1 the administered mice inhibitor transporters for intravenously investigate responsible and of strong primarily to exposure was (D6-M1) a aims blood CYP1A1 M1 and increased that to study indicated Deuterated Metabolic demonstrated experiments present vitro was recruited. The In models. were Gefitinib Results: vitro patients Key affected. in NSCLC M1. clearly using of Approach: than pharmacokinetic not investigated more Experimental was were by (NSCLC). gefitinib. ningetinib reduced mechanisms cancer was and lung of transport M1 ningetinib cell that metabolite of non-small surprising N-demethylated mechanism of of is interaction treatment exposure the it plasma for whereas high inhibitor the 80%, kinase gefitinib, tyrosine with a co-administered is When Ningetinib Purpose: and Background Abstract 2020 5, May 3 2 1 Xie hnhiIsiueo aei eia hns cdm fSciences of Academy Chinese Medica, Materia of Institute Sciences Ltd Shanghai of Co Academy Pharma Chinese Lake Medica Sunshine Materia of Institute Shanghai • • • • • 3 igLi Jing , osdrto o eaoiekntc nDIsuisapast ehlfli lcdtn h DDI the elucidating in helpful be to appears studies DDI in kinetics mechanism. demonstrated metabolite firstly CYP1A1. for was in which of resulted Consideration gefitinib, inhibitor efflux by canalicular strong inhibited was M1 a M1 on to patients of drug in formation parent the M1 the co-administrated, of of When exposure effect plasma inhibitory 80%. high the the and than affinity more tissue low by The reduced affected. was clearly M1 not of was ningetinib exposure of plasma pharmacokinetics drug. the expectations, parent to the gefitinib, Contrary of with that 1.7-fold co-administered about When was exposure plasma whose N 1 dmty igtnb(1 a dnie stepiaycruaigmtblt nNCCpatients NSCLC in metabolite circulating primary the as identified was (M1) ningetinib -demethyl inWang Qian , 2 n ioa Chen Xiaoyan and , 2 e Xie Cen , 3 igXi Ning , 3 2 ia Guo Zitao , 1 3 igLi Ming , 2 μ iny Hou Xiangyu , .C-diitaino ningetinib of Co-administration M. 3 Ningjie , Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. a,21;Rha ta. 04.I diin etnbwsas nihbtro h fflxtransporters efflux the of inhibitor an also Back- was & gefitinib Neuvonen, addition, In (Filppula, CYP2D6, degrees CYP2C19, 2014). varying inhibited pharmacokinetics al., to the gefitinib inter- et activities shown that drug-drug Rahman have note CYP2C8 a 2014; studies of to and suggestive Several man, interesting CYP1A2 80%, is than CYP3A4, unaffected. it more almost CYP2C9, Nevertheless, by was reduced circulating gefitinib. ningetinib was primary days and of the 28th ningetinib of and in between exposure evaluated first plasma firstly action the was the mg) on gefitinib, (60 pharma- M1 with ningetinib potential co-administered component and the safety, When mg) and (250 NSCLC. efficacy ningetinib. gefitinib with clinical of with patients the doses gefitinib on therapeutic of use of medication interaction co-administration concomitant cokinetic EGFR-TKIs as of combining such effect of NSCLC, the tolerability examine of and To Fu, treatment feasibility & the the (Huang explore in etc to PTEN), TKIs reasonable of multi-target (c-MET, loss be with pathways mutations, would (K-RAS alternative it pathway to of Therefore, downstream orphan due activation the inevitable 2015). granted Ranson of (T790M), is and aberrance mutation EGFR-TKIs 2014; and NSCLC secondary to the AXL) metastatic resistance Kassem, the HGF, and However, for as & Australia 2016). treatment such Korashy, al., mechanisms, first-line Japan, et various (Rahman, the (Kazandjian in FDA for NSCLC drug by approved chemoresistant qualification anti-cancer was drug of and an 2004) treatment as Wardell, the launched & agent for of EGFR-targeting studies Sates inhibitory clinical first United no in the concern almost is our had exposure. In cause M1 Gefitinib plasma should Although 2014). high it Wang, its drug. ningetinib, & parent of by Wu, the antagonised because Wang, targets of ningetinib antitumour that Zhang, the 1.7-fold Xi, on about 2018; phase effect was undergoing Jin, exposure now & plasma It‘s whose N-(3-fluoro-4-((7-(2-hydroxy-2-methylpropoxy)quinolin-4- (Wang. pathogenesis. AXL, treatment tumour as to NSCLC well relation as humans, for in VEGFR study (FLT3) c-MET, clinical 3 against kinase I/II (CT-053PTSA, (TKI) tyrosine inhibitor FMS-like kinase potent, and tyrosine Mer, molecule a small bioavailable is orally tosylate 1A) yl)oxy)phenyl)-1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazole-4-carboxamide,Fig astemizole, terfenadine, . Ningetinib as such to 2001). market, lead the use (Alfaro, even from the mibefradil drugs and drug-metabolizing However, of events and withdrawal NSCLC. of adverse cisapride, or of serious development induction cause of treatment icotinib or termination may the early with which inhibition for the unavoidable, apatinib to 2015) virtually of due al., transporters use interactions and/or et treatment combination enzymes cancer (Das drug-drug the of erlotinib makes as focus with polypharmacotherapy such the dovitinib of resistance become 2018), drug gradually al., overcome has et (Zheng and therapy drugs (Xia drug efficacy single-targeted combination the using et years, improve effect recent (Ai and to satisfactory In receptors etc achieve multiple 2017). of (BRAF), to interaction al., difficult B the et involve it homolog often makes mesenchymal- tumours oncogene which of pathways, (ALK), viral development signaling and sarcoma kinase occurrence murine The lymphoma v-Raf 2018). (KRAS), include anaplastic al., (c-MET), protein NSCLC (VEGFR), sarcoma factor for receptor rat transition human targets factor epithelial fac- kirsten therapeutic growth growth (PI3Ks), epidermal the 3-kinase epidermal targeted (mTOR), phosphatidylinositol And vascular molecular rapamycin (ErbB2), of emphasize 2 target patients. to receptor mechanistic of evolved tor Pisapia, (EGFR), have classification receptor Muscarella, factor NSCLC genomic (Malapelle, growth for stage the epidermal ma- advanced strategies the on late treatment and based a cancer, Currently therapy lung in of symptoms 2020). cases with Rossi, all present of & patients 80% approximately NSCLC for of accounts jority (NSCLC) cancer lung cell Non-small Introduction significance: Clinical • • h ogtr aeypol n ffiayo igtnbcmie ihgfiii hudb ocre in concerned be should gefitinib with combined ningetinib tumour of in efficacy patients. application and NSCLC its profile resistance. safety for tumour long-term possibilities terminal The provide of may reversal CYP1A1 and on prevention effect inhibitory strong Gefitinib’s N dmtyae igtnb(1 i B a dnie stepiaycruaigmetabolite, circulating primary the as identified was 1B) Fig (M1, ningetinib -demethylated 2 Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. oivsiaetemtblcsaiiyo igtnbadtefraino 1 rmr ua hepatocytes human primary M1, of formation the and ningetinib 37 of at incubated stability were metabolic 4 the at investigate centrifuged -70 To were 2, approximately samples 1, at blood 0.5, On stored the and were All days. administration EDTA-2K. samples 28 before containing 2.4 plasma for h tubes cm) The in 1 149–170.5 min. h within single of 10 24 with collected a height for and patients were trial, kg, 12 rpm the samples clinical 55.2–68.6 8, to blood of another 5, administered weight venous In 4, daily days, body 3, were 28. 28th old, old, of tablets day years and years mg) gefitinib to 47–76 first 28.5–45.9 (60 of 1 aged the aged dose mg day 4, 5, oral 250 was from = = single and (n cm) subject (n A capsules NSCLC 158–188 study ningetinib NSCLC study. of eligible of with the height Clinical dose patients Each in kg, oral to inclusion Good 51.9–73.8 S1. to h of of Table prior 24 weight principles form every in body consent ethical administered listed a was the are date capsules with criteria and ningetinib sign exclusion consistent read, and were Declaration, to inclusion studies Helsinki required the key clinical to The of According monitoring Center. Practice. Cancer and University Yat-sen design Sun the at enrolled were patients collection. The sample Ltd. and Co., Sciences treatment of Care Animal Subjects, Academy Animal Chinese Laboratory 2.3 the Medica, of Materia Sipper-BK principles of from Institute the Shanghai with purchased the China). accordance of were (Shanghai, in Committee male) Laboratory performed of were weight, Use studies and in animal All g China). (18–20 (Shanghai, mice ICR China). The (Jiangsu, University Soochow Pharmacy, of Animals. School provided 2.2 kidney the kindly were from canine a cells Zhang Madin-Darby using MDCKII-MRP2 Hongjian The G and generated Professor France). MDCKII-BCRP penicillin was by (Molsheim, cells), water trypsin, transfected system Deionised 0.25% (P-gp purification CA). (MDCKII)-MDR1 acids, water (Carlsbad, gradient DMEM amino Invitrogen Milli-Q (WME), non-essential from E Millipore purchased L-glutamine, Medium were (FBS), William’s streptomycin ob- serum China). sodium and was (Shanghai, bovine TAK-875 and MO). Ltd. foetal KO143 Louis, Co., (Kansas medium, GF120918, XenoTech Biochempartner (St. Shanghai Sigma-Aldrich Sekisui (NADPH), from from from phosphate purchased tained obtained were dinucleotide Research were (CMC-Na) Gen- adenine Disease hepatocytes cellulose BD Liver Nicotinamide carboxymethyl human Reid from Primary by obtained Kansas). supplied China). were were City, (Shanghai, (HLMs) (HLUMs) (HIMs), microsomes microsomes microsomes Ltd. liver CYP2E1, intestinal lung human Co., human Pooled human CYP2D6, MA). pooled and (Woburn, CYP2C19, and (HKMs) CYP450 test CYP2C9, recombinant CYP4F2) microsomes Human and kidney CYP2C8, CYP4A11 human China). CYP2B6, were CYP3A5, vincristine (Liaoning, CYP2A6, CYP3A4, and CYP2J2, Ltd. gefitinib CYP1A2, provided imatinib, Co., kindly (CYP1A1, Digoxin, Biotechnology were China). enzymes Meilun (D6-M1) (Guangdong, Dalian M1 Ltd. from deuterated Co., purchased Pharma and Lake (D6-ningetinib) Sunshine ningetinib by deuterated M1, Ningetinib, reagents. and M1 Chemicals effect and 2.1 ningetinib the on and effects formation different Methods M1 the and for of Materials reasons mechanism of the variety the reduced. explore a investigate the dramatically further using to of was and gefitinib was concentration exposure production, by study M1 plasma plasma present on high the the gefitinib a that of of with puzzling M1 purpose gefi- was of the by that it al., transporters Hence, when and Besides, et changed enzymes Kitazaki hardly metabolizing M1. 2015a; has these of drug al., of pharmacokinetics parent et inhibition the whether (Galetti unknown affected (BCRP) was tinib protein it resistance date, To cancer 2005). breast and (P-gp) P-glycoprotein rHUswr nuae t05m/Li 0 Mpopaebffrcnann . MMgCl mM 3.2 containing buffer phosphate mM 100 in mg/mL 0.5 at incubated were HLUMs or nvitro In eaoi tblt experiments. stability metabolic ° nWEa 1.0 at WME in C nvitro in and × 10 nvivo in 6 el/Lcnann 3 containing cells/mL eaoi n rnpr models. transport and metabolic 3 μ igtnb h Ls Is HKMs HIMs, HLMs, The ningetinib. M ° ni analysis. until C 2 ° PS pH (PBS, t2000 at C Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. oiiesbtae n icitn MP oiiesbtae eeue o aiaino h bv efflux above the of validation for used were substrate) positive (MRP2 vincristine and for incubated substrate) were -20 equilibrium. positive cells at the The stored of chambers. were end both samples to the 37 added inhibitors at at were positive removed min inhibitors 90 of was positive the absence HBSS and or Equilibration chamber), (10 presence basolateral 2015)]. ningetinib the Chen, in containing HBSS & min HBSS prewarmed Zhong, 30 with Guo, for twice Zhong, equilibrated (Li, washed were were cells chambers (10 the GF120918 basolateral then, and and study. 7.4), apical MDCKII-MDR1/BCRP/MRP2 transport (pH The bidirectional the day. the assessment, other before substrate every days For one 4 fresh for grown a were with lines replaced cell routinely was medium The cells/cm h DKIMR/CPMP el eeclue nDE upeetdwt 0 B,2m L- mM 2 37 FBS, 10% at 100 with acids G, supplemented amino penicillin DMEM U in 100 cultured glutamine, were cells MDCKII-MDR1/BCRP/MRP2 The ies. -20 was at stud Plasma stored Transport were paper. 2.7 samples filter All with min. 10 weighed wiping for blood, after g or 11,000 v/v) contents at analysis. (1:1, out blood acetonitrile-water rinse of to centrifugation of pancreas, by saline volumes bladder, obtained by gland five kidney, 2, washed adrenal lung, in were 0, fat, spleen, tissues homogenised abdominal at liver, The intestine, and anaesthesia collected. heart, large intestine, under rapidly brain, small were aorta blood, thymus wall, in The stomach abdominal and testis, administration. dosage the limb), after Each clinical from (hind h formula.). muscle the exsanguination 48 skeletal Meeh-Rubner via to or the equivalent 24 sacrificed to 12, was basically according 6, with mice was differences h kg three mice interspecies 12 mg of in for 10 for group fasted here at corrected were ningetinib used when which of patients dose groups, administration NSCLC six ningetinib oral into single the divided a solution; randomly received were CMC-Na and mice water ICR to access male samples mice. healthy free incubation 18 in All of M1 initiation. total and A reaction’s ningetinib the of after distribution min 60 Tissue and 2.6 -20 30 at 60, stored 15, and 5, triplicated 100 at were was reaction mixture the reaction terminate each to of volume The mM). 100 pmol containing 10 mixture or reaction 5 mg The 10, (0.05 selected. (5, HLMs were CYP1B1, CYPs or range CYP1A1, with respectively) in linear supplemented CYP3A4, incubation explored of 7.4) the and or was concentration (pH metabolism in enzyme M1 PBS the conditions the of mM catalyze of incubation formation optimisation could the the After CYP3A4 incubations. on time, and HLMs inhibition and CYP2C9 gefitinib CYP3A4 CYP1B1, The CYP2C9, CYP4F2 CYP1A1, and M1. The CYP4A11 into CYP2B6, work. CYP3A5, ningetinib CYP2A6, CYP3A4, previous CYP1B1, CYP2J2, our CYP1A2, CYP2E1, CYP1A1, in CYP2D6, using evaluated CYP2C19, was CYP2C9, formation CYP2C8, gefitinib. M1 by of kinetics phenotype -20 inhibition enzyme at of stored determination and i and triplicated The production were M1 samples of incubation phenotype the Enzyme All 2.5 reactions. the of initiation ° the after min 180 100 (3 ningetinib and 7.4) diin ieetcnetain fnneii eeaddt h Y11(.,03 .,27o 8 or 2.7 0.9, 30 0.3, or (0.1, 10 6, CYP1A1 3, the (1, 1 to mixtures or reaction 0.5 added HLMs 0.2, were 0.1, ningetinib (0.05, of CYP1B1 concentrations different addition, o CM/Sanalysis. LC-MS/MS for C μ vitro n .Teratoswr emntdb digeulvlm fiecl ctntiea ,5 5 0 0and 60 30, 15, 5, 0, at acetonitrile ice-cold of volume equal adding by terminated were reactions The L. 2 naplcroaemmrn le ebaeo rnwl net Mlioe ilrc,MA). Billerica, (Millipore, inserts Transwell on membrane filter membrane polycarbonate a on eaoimeprmnssoe htM a anyfre nlvrmcooe.Teeoe the Therefore, microsomes. liver in formed mainly was M1 that showed experiments metabolism ° ,a hc ieaiut (150 aliquots time which at C, μ ,Pg niio) O4 (10 KO143 inhibitor), P-gp M, ° nahmdfid5 CO 5% humidified a in C μ )i h rsneo AP 2m) h oueo ahicbto itr was mixture incubation each of volume The mM). (2 NADPH of presence the in M) ° o CM/Saayi.Dgxn(-ppstv usrt) mtnb(BCRP imatinib substrate), positive (P-gp Digoxin analysis. LC-MS/MS for C μ μ ° ) Y29(,6 2o 30 or 12 6, (3, CYP2C9 M), )o 1(10 M1 or M) eoeL-SM analysis. LC-MS/MS before C μ mL g -1 μ ) h ecin eeiiitdb h diino AP (2 NADPH of addition the by initiated were reactions The M). tetmcn n %mnmmesnilmdu nonessential medium essential minimum 1% and streptomycin, μ )wr olce rmtercie hmesfraayi.The analysis. for chambers receiver the from collected were L) 2 · protein μ topee h el eesee tadniyo 2 of density a at seeded were cells The atmosphere. ,BR niio)o A85[100 TAK875 or inhibitor) BCRP M, μ )wsaddt h oo ie(ihrteaia or apical the (either side donor the to added was M) μ 4 .A qa oueo c-odaeoirl a added was acetonitrile ice-cold of volume equal An L. · mL -1 a nuae ih0.01–30 with incubated was ) μ ) Y34(,5 5 0o 60 or 30 15, 5, (3, CYP3A4 M), · mL -1 fCPA,CPB,CYP2C9 CYP1B1, CYP1A1, of -1 μ ° ,MP inhibitor MRP2 M, sseddi 0.5% in (suspended ni LC-MS/MS until C μ etnb In gefitinib. M μ )and M) × μ M), 10 5 Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. 17)mtos h ierrgeso ie bandi u xeiet eealitretda n point. (V one = at v intersected inhibition Cornish-Bowden all mixed-type inhibitions. and were the mixed-type (1953) experiments by and Dixon fitted our of well in Prism were basis obtained Graphpad [S](1+[I]/ data the the lines inhibition On using regression the model CA). linear nonlinear Thus, inhibition Jolla, by the enzyme La determined best methods, Inc., the accurately (1974) Software of more GraphPad basis were the 8.01; and on (version methods, analysis (1974) regression square Cornish-Bowden least and (K (1953) activity (CL Dixon inhibitory clearance for as parameters Intrinsic kinetic apparent equation. to The Michaelis–Menten used the was V using NC) studies, fit Cary, kinetics inhibition Corp., model. the Pharsight noncompartmental For a 6.1; in (version parameters software pharmacokinetic WinNonlin the al., study, calculate et pharmacokinetic (Curtis pharmacology the in In analysis anal- and statistical design and experimental data on The recommendations 2018). designed. the double-blind with were comply analysis yses statistical the and All procedures heater Experimental gas, respectively. nebuliser psi, The 20 ionisation degC. electrospray and 400 analysis. positive 50 of Data acetonitrile. the temperature 50, with in source to min, precipitation used a set 2.5–3 was protein and min; were D6-M1) by D6-ningetinib, V 2.5 for for prepared gas 4500 at 215.3 271.5 curtain of B - - voltage and 20% 563.4 549.4 spray gas to m/z m/z ion decrease ningetinib, and an linear for M1 with stepwise 215.0 for mode a - 271.1 B; 557.3 - 80% (m/z 543.1 min, monitoring m/z 1.5–2 reaction min; Multiple 1.5 B. at 20% min B mL 80% 0.6 to of increase flow Chroma- a US). at MA, (B) mm 40 Sciex, (50 (AB at C18 software Japan) YMC-Triart 1.6.3 Bldg, on Analyst achieved using was conducted separation Japan). was spectro- tographic Kyoto, mass processing (Shimadzu, quadrupole system and triple chromatography acquisition 5500 liquid API Data high-performance an LC-30AD using an after with conducted was coupled h M1 meter 48 and and ningetinib 24 of analysis 12, Quantitation 8, preparation. 6, sample 4, -20 and 2, at 1, Instruments stored 0.5, were 2.9 0.25, samples 0.083, Blood and h) D6-M1. (0 of administration administration before solution mL mg citrate (10 (0.5 trisodium solution D6-M1 CMC-Na later 0.5% min of 30 (5 volume of Then, same concentration administered. the mg orally blood then, group, was (40 and control initial advance, ningetinib the in high group, In min the treatment 30 injection. gavage considered the by mg mice In design (0.5 to experiments. dose D6-M1 administrated the was This intravenously.) before administered solution; when fasted water CMC-Na D6-M1 were to They 0.5% access groups. in control free and suspended with treatment into mice. h divided in 12 randomly D6-M1 were for mice of ICR pharmacokinetics male healthy the Fourteen on evaluated. ningetinib further effects of ningetinib were the by Effect MRP2 replaced plasma, 2.8 and were the inhibitors BCRP positive in P-gp, the by drug whereas mediated parent above, M1 as the (0–100 same of than the efflux were slowly the procedures more on Experimental eliminated gefitinib was and ningetinib M1 of metabolite the that Given system. μ o ah eecletdtruhteti enadpae nts ue otiig30 containing tubes test in placed and vein tail the through collected were each) for L μ )o etnb(0–100 gefitinib or M) α K i .Temdl etdicue ueadprilcmeiie ocmeiie uncompetitive noncompetitive, competitive, partial and pure included tested models The ). · kg -1 isle nsln otiig5 MOad5 we 0 a ie hog alvein tail through given was 80) Tween 5% and DMSO 5% containing saline in dissolved , ° .Tembl hs a itr f5m moimaeae()adacetonitrile and (A) acetate ammonium mM 5 of mixture a was phase mobile The C. -1 h rdetcniin eea olw:2%Bfr05mn tpielinear stepwise a min; 0.5 for B 20% follows: as were conditions gradient The . μ ) l ape eesoe t-20 at stored were samples All M). max n K and m auswr eemndb h olna ersincurve regression nonlinear the by determined were values 5 i eefis siae ygahclmtossuch methods graphical by estimated first were ) · kg ° ni CM/Sanalysis. LC-MS/MS until C -1 × a netditaeosy lo samples Blood intravenously. injected was ) int . mid,5 i.d., mm 2.0 ° a acltda CL as calculated was ) eoeL-SM analysis. LC-MS/MS before C nvitro in max μ and ;YCKarasuma-Gojo YMC m; · S) (K [S])/ kg -1 nvivo in ihu ningetinib without ) m int 1+[I]/K + (1 ape were samples V = μ f01M 0.1 of L max · /K kg i + ) -1 m , . Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. h eaoiepol fnneii nmc a iia ota fteNCCptet TbeS n Figure and S2 (Table because patients mg mice NSCLC 10 in for the administering determined explanation of after was that plausible 5A, M1 to a similar and Fig. provide was ningetinib to To mice of According in distribution ningetinib humans. S1). tissue of in the profile ningetinib metabolite M1, of the of pathway exposure metabolic plasma primary high the for not accounted was excreta in M1 mice. that in M1 vitro and gefitinib. In ningetinib by inhibited of not distribution on was Tissue effect incubation K determined 3.4 inhibitive CYP3A4 was with the the an type nature and in inhibition demonstrated inhibitory M1 HLMs The gefitinib competitive of incubations. on 4, mixed-type production CYP2C9 gefitinib Fig. a and of in CYP1B1 be kinetics shown CYP1A1, to inhibition the As HLMs, The studied. in to formation effect. further According CYP1A1, M1 was catalytic M1. examined, enzymes minor into enzymes P450 a ningetinib P450 four had of of CYP3A4 metabolism kinds and the 15 CYP2C9 catalyze the could among CYP3A4 gefitinib. that CL by showed and kinetics laboratory CYP2C9 inhibition our CYP1B1, A and in 3). production study (Fig M1 Previous microsomes for liver responsible the enzymes in P450 mainly 3.3 consumed, was ningetinib of HLUMs. ( and 10% M1 than of amount less by small min, explored initially 180 was for both formation bation M1 of of vitro. behaviours mechanism in The elimination formation The M1 day. of gefitinib, and 28th Investigation ningetinib 3.2 and between DDI mechanism. first a suggested metabolic the data through These on likely changed. obviously mostly 80% not co-administration were than After M1 ningetinib. and more ningetinib of by that than dropped slower much (T significantly C was the concentration M1 gefitinib, of peak rate with the elimination the reach and (C to concentrations ningetinib, peak time the AUC alone, the and given comparable, Moreover, was were ningetinib plasma when in 1, NSCLC. Table M1 and with and 2 patients Fig in in shown gefitinib As with ningetinib of interaction 3.1Pharmacokinetic GraphPad in t-test Results student’s unpaired the using 10 evaluated + were Prism. 100/[1 groups two = between Y comparisons follows: Statistical as response normalised IC from the and the against study, sides concentration basolateral inhibition to transport apical the the In P cm respectively. from time. sides, transport (0.33 incubation apical the monolayer = by to T cell generated basolateral and permeation the the side of donor of extent the the on area represent compound surface test the the of = concentration S side, receiver C where P = ER P (P formula: coefficients following permeability the apparent using the lated experiments, transport the In app int C = aussoni al ,CPA lydapeoiatrl nM omto,weesCYP1B1, whereas formation, M1 in role predominant a played CYP1A1 2, Table in shown values P p,Bt A to B app, T xeiet hwdta 1frainwsls hn5 nHLMs. in 5% than less was formation M1 that showed experiments T < h ocnrto ftets opudo h eevrsd,V=telaigvlm nthe on volume loading the = V side, receiver the on compound test the of concentration the = × .5wscniee significant. considered was 0.05 /(C V/ P / 0 max p,At B to A app, < × % a eetdi Lsadee esi Is 1wsntdtce nHKMs in detected not was M1 HIMs. in less even and HLMs in detected was 5%) T n AUC and × S) < 0-24h %o h oeamnsee ute upre that supported further administered dose the of 2% fnneii eeams nhne,weestoevle o M1 for values those whereas unchanged, almost were ningetinib of 50 auswr acltdb ltigtelgvleo h inhibitor the of value log the plotting by calculated were values 0-24h au fM a prxmtl .-odta fningetinib. of that 1.7-fold approximately was M1 of value 6 · kg -1 max i fnneii rlyt h ie h aetdrug parent the mice, the to orally ningetinib of nvitro in f1.,005 .3ad9.23 and 1.83 0.095, 16.4, of fM a infiatylne hnta of that than longer significantly was M1 of ) eaoi nuainsse.Atrincu- After system. incubation metabolic 2 na2-elpae,C plate), 24-well a in app n fflxrto(R a calcu- was (ER) ratio efflux and ) (X-Log(IC50) nvivo In p,At B to A app, xeietlevidence experimental ]. μ N max .Hwvr the However, M. n P and -demethylation 0 fningetinib of ) h initial the = p,Bt A to B app, Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. omto a anymdae yCPA n oalse xetb Y11 Y29adCYP3A4. and CYP2C9 CYP450s CYP1B1, M1 of by that inhibitor extent showed an lesser data as a kinetic gefitinib to enzymatic of and The potency CYP1A1 the investigated. by examining initially mediated When mechanism. was mainly metabolic formation was through These M1 formation likely changed. mostly of obviously gefitinib, not mechanism were and The M1 ningetinib and between ningetinib DDI both a of C suggested behaviours elimination (AUC, data The parameters affected. pharmacokinetic not the were whereas gefitinib, plasma with humans. the administered both in therapy, ningetinib NSCLC of C for medication metabolite and combination circulating exposure further multi-target-based primary on which the trials 200%, clinical as in identified by However, was nearly M1 studies, increased pharmacokinetic 4 group In From ningetinib the administration. blood D6-M1. after in the of h D6-M1 elimination group, Discussion 2 of the control within inhibit exposure the addition, could ningetinib blood with ningetinib In by the Compared that affected 75%. demonstrated h, not approximately trans- 4. 24 by was Table efflux to ningetinib increased D6-M1 and by was h of inhibiting 7 generated group concentration by M1 ningetinib-combination blood Fig. of the M1 the effect in in the D6-M1 shown of prevent of are to pharmacokinetics exposure results mice the ICR The to affect injected metabolism. intravenously could was mice. D6-M1 ningetinib The in porters, whether D6-M1 exposure. ningetinib. of investigate plasma of pharmacokinetics To that its on than elevate ningetinib weaker thereby significantly of could and was Effect ningetinib M1 efflux 3.6 drug M1 metabolite parent on its the gefitinib of of that efflux effect indicated inhibitory canalicular phenomenon, the inhibition inhibit P-gp potently the IC especially with results, efflux These M1 BCRP-mediated 9.09 and and P-gp- on inhibition concentration-dependent h oto ru)wspotdo h riae ssoni i.6 igtnbdslydaconcentration- in a IC that displayed with to Ningetinib efflux group 6. M1 inhibitor Fig. the MRP2-mediated in the shown in and and as ratio BCRP- abscissa, ordinate, efflux P-gp-, μ the the of on on on (percentage inhibition plotted plotted MRP2 further dependent was was was or group) efflux gefitinib BCRP control M1 or P-gp, the MRP2-mediated ningetinib and of BCRP of activity P-gp, concentration remaining on MRP2 logarithm gefitinib BCRP. and The and ningetinib P-gp investigated. of of ningetinib. effect substrates of inhibitory were not except The M1 but and result M1 ningetinib similar of both efflux a KO143 Thus, the displayed inhibitor), cells. mediated ningetinib (P-gp apart MDCKII-MRP2 MDCKII-MDR1, while in 2 GF120918 in M1 respectively, than that of of cells, for values greater addition ER all MDCKII-MRP2 the the were reduced and Then stable- markedly ningetinib/M1 MDCKII-BCRP inhibitor) cells. cells of (MRP2 TAK875 MDCKII-MRP2 values MDCKII and in ER inhibitor) the (BCRP The ningetinib (MDCKII-MRP2) transporters, 3). of MRP2 (Table efflux that or screening of from (MDCKII-BCRP) substrate substrate for BCRP on a used (MDCKII-MDR1), transporters was were P-gp efflux M1 human of or with influence overexpressed ningetinib The studied. blocked. whether further patients, be determine was NSCLC to To in M1 likely and and was mice ningetinib ICR M1 of in of drug disposition pathway parent the transporters. efflux the efflux the than half-life site by because elimination preferred mediated possibly plasma the M1 longer was a on in Plasma exhibited gefitinib M1 concentrations 5C). and (Fig. M1 ningetinib drug the of parent However, in the Effects drug. that gradually of 3.5 parent than mice 30% vivo. the lower than in in less double all M1 disposition were were than of M1 liver liver for more concentration the the even plasma from than was the apart plasma. tissues other and pharmacokinetics, in human tissues time than over to in tissues most increased Similar concentrations in 5B). M1 exposure (Fig. the higher plasma drug, a parent with distribution the tissue Unlike extensive an exhibited ningetinib ) 18.7 M), μ (0–30.0 M μ (0–100 M max fM rpe rmtclyadwr eue ymr hn8%we igtnbwsco- was ningetinib when 80% than more by reduced were and dramatically dropped M1 of μ ) epciey u hwdn niiino R2mdae 1eu (0-100 efflux M1 MRP2-mediated on inhibition no showed but respectively, M), μ )ad3.06 and M) μ (0–30.0 M μ ) epciey iial,gfiii xiie weak a exhibited gefitinib Similarly, respectively. M), 7 nvitro in u eut eesomewhat were results our , max 50 n T and auso .1 (0–100 0.413 of values 50 f54 (0–100 5.40 of max fningetinib of ) μ μ M). M) Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. u oterltvl ihepeso fCPA nploaytmu ise,tecnetaino the of concentration the tissues, tumour pulmonary the M1. in However, of CYP1A1 yield 80%. metabolic of than low expression the more of high by CYP1A1, because high relatively exposure of influenced its plasma the not inhibitor in was its to strong resulted ningetinib reduced a Due drug efflux and parent as canalicular M1 the M1 co-administrated, of of on When pharmacokinetics formation ningetinib the half-life. of inhibited elimination effect gefitinib long inhibitory the and NSCLC. with and exposure of patients M1 plasma amount in of small gefitinib a affinity and produce tissue an ningetinib to low of as CYP1A1 mechanism by potential DDI metabolized its the was provide Ningetinib illustrated from may study HLUMs this CYP1A1 and conclusion, for In HLMs ningetinib in of CYP1A1 selectivity of CYP1A1. high gefitinib’s expression and of The protein prevention indicator Therefore, degrees. the tumour varying with in 2020). of correlated application smokers al., positively its et was positively for produced (Lu were possibilities resistance. M1 et,al provide tumour rs1048943) may terminal cancer, and of CYP1A1 lung (rs4646903 reversal on cancer, polymorphisms re- effect breast CYP1A1 also inhibitory Mahjabeen, cancer, Wahid, was addition, strong 2013; It prostate In al., with 2008). et the 2013). al., (Androutsopoulos correlated to compared et expression Kayani, tissues level (Oyama tumour & mRNA cancer bladder by Baig, and lung colon determined CYP1A1 of brain, as human CYP1A1. development tissues, in of hydrocarbons the normal expressed inhibitor aromatic highly in was strong polycyclic role CYP1A1 a in that a involved was ported play enzyme gefitinib 50 CYP might that pulmonary of and report human effect metabolism, to studied time actively the ignored. work. first most under be present the the 50% could was was the ningetinib than it in on knowledge, more studied gefitinib our The not To also of transports. by effect was efflux decreased transporter-mediated BCRP its were Therefore, on and IC ningetinib not S3). P-gp higher (Table formation, of by its M1 values mediated to on ER gefitinib gefitinib due was The and of M1 negligible ningetinib effect of was inhibitory pathway of gefitinib the efflux interaction showed the by M1 that efflux of speculated of alteration was lower inhibition it much C situation, the plasma and this The whereas value In patients. ningetinib, ningetinib. NSCLC by of to third inhibited administered one were (1.37 than drugs ningetinib less these of was (9.09 of concentrations ningetinib doses plasma of therapeutic peak that when than the lower than was M1 higher BCRP of much and efflux on 2005) the gefitinib al., inhibit of might et value It Kitazaki BCRP. (5.40 and IC 2015b; and ningetinib P-gp the al., However, of P-gp of BCRP. et inhibitor and of an (Galetti experiment P-gp also inhibitor pharmacokinetic literature by was D6-M1 mediated potent the gefitinib The that biliary addition, a confirmed the exposure. In results be obstructed plasma our ningetinib high conclusion. to its that this happened in speculated proved substrate resulted was ningetinib further a and it drug was M1 Therefore, of parent M1 MRP2. pathway that The and excretion BCRP demonstrated of MRP2. studies inhibitor transport and moderate The BCRP subsequently. P-gp, evaluated of in was both M1 rates of 90%). binding pathway versus protein metabolite (99.9% higher the drug ningetinib. much vitro parent of that exhibited In that the suggesting M1 than than difficult drug, that plasma more was parent was human findings the tissues plasma, and into the of in mouse penetration for Unlike 30% its explanation and than clearance. plasma possible and/or less the One distribution in were retained tissue tissues be to in to related tended M1 be of could M1 concentrations of the the and exposure since 5%, plasma gefitinib, than high by less The influenced humans. was not patients in was inhibited in of ningetinib al., drug significantly and of inhibitor parent Gefitinib et HLMs pathway moderate the in metabolic CYP3A4. Rahman of CYP1A1, produced on concentration of M1 2014; influence the of inhibitor no but Backman, amount strong had HLMs, & but a in CYP2C9 was production Neuvonen, of M1 gefitinib (Filppula, inhibitor that weak literature revealed and study the CYP1B1 present in The reported 2014). those from different nuain hwdta h ie a h rmr ieo 1frain hs h eai clearance hepatic the Thus, formation. M1 of site primary the was liver the that showed incubations μ ess0.413 versus M nvivo in ocnrtos hnc-diitae ihgfiii,tepharmacokinetic the gefitinib, with co-administrated When concentrations. μ )adgfiii a oihbtr ffc nMP.Tog h IC the Though MRP2. on effect inhibitory no had gefitinib and M) nvitro In 50 xeiet norlbrtr niae htteaon of amount the that indicated laboratory our in experiments au fgfiii nPg a infiatyhge hnthat than higher significantly was P-gp on gefitinib of value 8 μ ess18.7 versus M N μ g N dmtyae eaoieM.The M1. metabolite -demethylated · mL dsehlto a o major a not was -desmethylation -1 n etnb(7 ng (272 gefitinib and ) μ ) oho hmwere them of both M), μ max fgefitinib of M fgefitinib of · mL -1 50 50 ) Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. nteteteto o-ml elln acr urn hlegsadtewyforward. alterations way molecular the emerging and Targeting challenges current (2020). cancer: A. lung Rossi, cell endogenous & non-small in P., of CYP1A treatment Pisapia, of the A., in insights L. New Muscarella, diseases. U., (2020). and Malapelle, X. Wang, polymorphisms & nucleotide R., single Shi, on Y., doi:10.1016/j.apsb.2019.11.016 Hepatobiliary focus Xu, Inhibits a W., (TAK-875) Zhong, metabolism: Fasiglifam X., Shang, (2015). J., X. Lu, Injury. Chen, Liver Fasiglifam-Induced & doi:10.1124/dmd.115.064121 to D., Contributing 1751-1759. Zhong, Factor (11), resistant Z., Possible multidrug Guo, A in K., Transporters: Gefitinib, P-glycoprotein Zhong, (2005). of S. X., Kohno, function Li, the . inhibits . directly . M., inhibitor, cells. Yasunaga, cancer S., kinase Approval Doi, tyrosine J., FDA Lung EGFR Tsurutani, Cell Y., an (2016). Nakamura, Non-Small M., R. Mutation-Positive Oka, Pazdur, T., EGFR & Kitazaki, Metastatic P., with Keegan, Patients K., of Treatment He, Cancer. the W., Yuan, for Gefitinib M., of inhibitors. G. R. kinase Blumenthal, R. tyrosine D., Alfieri, EGFR Kazandjian, to . resistance of . Mechanisms Lines. 5 (2015). . Cell L. B, M., Fu, NSCLC & in Bonelli, L., Gefitinib S., Huang, of R. Monica, Uptake R. and La Alfieri, Efflux D., on 10 Cretella, . Expression One, C., ABCG2/BCRP . of Fumarola, Effect G., . P. (2015b). Lines. M., Cell Petronini, NSCLC Bonelli, M., in S., Gefitinib Galetti, Monica, of Uptake La and D., Efflux on 10 Cretella, Expression One, C., ABCG2/BCRP Fumarola, of Effect G., P. (2015a). Petronini, M., inhibitors. inhibitory kinase Galetti, time-dependent protein of fourteen assessment vitro by In activity doi:10.1124/dmd.114.057695 CYP3A (2014). 1202-1209. T. and J. CYP2C8 Backman, on & J., effects Dovitinib P. Neuvonen, interaction. (2015). M., drug-drug A. A. A H. Wakelee, Filppula, cancer: & W., lung J. cell for Neal, non-small W., guidance Ahluwalia, metastatic J. with 89 simplified Riess, . patients L., and in Zhou, . erlotinib updated A., Frymoyer, . and K., II: A., S. reporting M. Padda, M., Giembycz, their Das, H., and C. analysis reviewers. George, and peer R., and design J. Docherty, authors Experimental . G., Cirino, tumors. (2018). . S., bladder A. and Alexander, . colon J., D., in M. Delakas, enzymes Curtis, CYP1B1 M., and Kyriakakis, CYP1A1 E., of 8 A. profile One, Papalampros, Expression and PLoS A., (2013). bad, Ploumidis, M. the A. I., good, Tsatsakis, the Spyrou, P., development: drug V. therapies in Targeted Androutsopoulos, studies (2018). interaction S. drug Zhu, of & role unknown. D., Emerging the Zhang, (2001). N., L. M. C. P. Alfaro, Joslin, cancer. L., lung A. cell Stancu, non-small J., advanced Wang, for X., Guo, X., or Ai, enzymes studies metabolic DDI of in inducers) patients. (or metabolites inhibitors NSCLC Reference of novel in pharmacokinetics of long-term concerned discovery the and the be of mechanism consequence, should DDI analysis transporters. a the gefitinib in-depth As enrich with that help gefitinib. combined suggested can by ningetinib also increased of findings be may efficacy The tissue and target profile the safety in ningetinib drug parent 3,2026 doi:10.1016/j.lungcan.2015.06.011 280-286. (3), 5,3041 doi:10.1016/j.apsb.2015.07.001 390-401. (5), lnclCne eerh 22 Research, Cancer Clinical (11). doi:10.1371/journal.pone.0141795 e0141795. (11), ugCne,49 Cancer, Lung 1) 847 doi:10.1371/journal.pone.0082487 e82487. (12), scohrao ul 35 Bull, Psychopharmacol rJPamcl 175 Pharmacol, J Br 3,3733 doi:10.1016/j.lungcan.2005.03.035 337-343. (3), 6,1307-1312. (6), noagt 9 Oncotarget, 4,80-93. (4), 7,9793 doi:10.1111/bph.14153 987-993. (7), 9 11,359367 doi:10.18632/oncotarget.26428 37589-37607. (101), caPamSnB 10 B, Sin Pharm Acta rgMtbDso,42 Dispos, Metab Drug rgMtbDso,43 Dispos, Metab Drug xetOi Investig Opin Expert caPamSin Pharm Acta ugCancer, Lung 1,91-104. (1), PLoS Plos (7), Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. xeiet.Rmiigatvt en ecnaeo Rvle fM nteihbtrgopvru the mg versus 40 without group experiments or inhibitor with the D6-M1 mg/kg in independent M1 7 three of of Fig. values mean ER the of represents percentage point group. Each means control cells. activity independent (C1/C2) Remaining three II-MRP2 experiments. of MDCK mean and the (B1/B2) represents BCRP point 6 Each ratio. Fig. ningetinib to M1 (C) experiments M1, (B) ningetinib, (A) figures experiments 5 The independent Fig. three method. of the (1974) mean Cornish-Bowden represent the the represents above point represent Each figures middle ± inhibition. the The mixed-type in the (D1/D2/D3). represent figures below CYP2C9 The method. or (1953) (C1/C2/C3) Dixon CYP1B1 experiments (B1/B2/B3), independent kidney CYP1A1 three human of and 4 mean (D) Fig. the represents microsomes point lung Each human time. (C), ± incubation microsomes gefitinib. versus intestinal of (E) mg human microsomes 250 (B), (B1/B2) microsomes with liver and (A1/A2) 3 without ningetinib Fig. of mg 60 of administration Anticancer oral Multi-Targeted 2 (2017). Fig. Y. F. 1 Wang, & Fig. K., Wu, Figures Y., for Legends Zhang, Q., icotinib Luo, with c-MET Y., apatinib EGFR Agents. novel of Zhao, with therapy a W., adenocarcinoma Combination pulmonary CT053PTSA, Zheng, (2018). advanced J. with Hu, (2014). patients & in J. W., resistance Lv, mutation. T. icotinib L., Wang, acquired Wang, X., primary & Lv, for growth. J., J., tumor Cao, Y. P., and Xia, Wu, trials. and angiogenesis clinical H., suppresses CYP1A1 in Z. potently inhibitors Wang, of doi:10.1158/1538-7445.Am2014-1755 inhibitor, c-Met J., Expression of VEGFR2 Y. progress and Zhang, research N., (2013). The Xi, A. (2018). M. R. Kayani, Jin, 20 Research, Pakistan. & & Y., in M., Wang., tissues R. tumor Baig, brain doi:10.7314/apjcp.2013.14.12.7187 human I., cancer. in of treatment Mahjabeen, GSTP1 the for M., (2008). agent administered T. Wahid, orally Kawamoto, 29 novel, Gefitinib. a Ther, Gefitinib, Pharm . (2014). Clin (2004). S. J G. Wardell, . M. & M., Kassem, . Ranson, & H., M., Uramoto, H. N., 39 Korashy, Methodol, Nose, F., cancer. lung A., A. cell Matsumoto, Rahman, non-small in T., P450 Isse, cytochrome of K., Expression Sugio, T., Oyama, Drugs E n=3.“”masgfiii n S en ningetinib. means “S” and gefitinib means “I” 3). = (n SEM 3). = (n SD -0 doi:10.1080/13543784.2020.1732922 1-10. , niioyeeto etnbo 1frainmdae yhmnrcmiatHM (A1/A2/A3), HLMs recombinant human by mediated formation M1 on gefitinib of effect Inhibitory isedsrbto fnneii n 1i C ieatra rlds fnneii 1 mg (10 ningetinib of dose oral an after mice ICR in M1 and ningetinib of distribution Tissue urn oisi eiia hmsr,17 Chemistry, Medicinal in Topics Current tutr fnneii A n 1(B) M1 and (A) ningetinib of Structure niioyeeto igtnbadgfiii nM fflxi DKIMR A/2,MC II- MDCK (A1/A2), MDCKII-MDR1 in efflux M1 on gefitinib and ningetinib of effect Inhibitory enbodcnetaintm rfie fD-1i iefloigitaeosijcino 0.5 of injection intravenous following mice in D6-M1 of profiles concentration-time blood Mean eaoi tblt fnneii n omto fM nhmnpiayhptcts() human (A), hepatocytes primary human in M1 of formation and ningetinib of stability Metabolic enpam ocnrto-iepolso igtnbadM nptet ihNCCfollowing NSCLC with patients in M1 and ningetinib of profiles concentration-time plasma Mean hrcCne,9 Cancer, Thorac ± ± 524-528. , 3-6.doi:10.1016/B978-0-12-800173-8.00005-2 239-264. , D( ) p * 7). = (n SD 3). = (n SD 2,9-0.doi:10.1111/j.1365-2710.2004.00543.x 95-103. (2), 5,6661 doi:10.1111/1759-7714.12624 656-661. (5), < .5 *p ** 0.05, · kg -1 igtnb ahpitrpeet h eno he independent three of mean the represents point Each ningetinib. < .1 * p *** 0.01, 2) 3084-3098. (28), 10 sa a acrPe,14 Prev, Cancer J Pac Asian < .0 s h control the vs. 0.001 rn isi 13 Biosci, Front rfie rgSbtEcpRelat Excip Subst Drug Profiles 5787-5793. , acrRsac,74 Research, Cancer 1) 7187-7191. (12), acrCell Cancer · kg (19). -1 ). Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. lung-cancer-patients of-ningetinib-and-gefitinib-involving-cyp1a1-and-efflux-transporters-in-non-small-cell- 4.docx Table file Hosted lung-cancer-patients of-ningetinib-and-gefitinib-involving-cyp1a1-and-efflux-transporters-in-non-small-cell- 3.docx Table file Hosted lung-cancer-patients of-ningetinib-and-gefitinib-involving-cyp1a1-and-efflux-transporters-in-non-small-cell- 2.docx Table file Hosted lung-cancer-patients of-ningetinib-and-gefitinib-involving-cyp1a1-and-efflux-transporters-in-non-small-cell- 1.docx Table file Hosted vial at available at available at available at available https://authorea.com/users/303604/articles/433788-drug-interaction- https://authorea.com/users/303604/articles/433788-drug-interaction- https://authorea.com/users/303604/articles/433788-drug-interaction- https://authorea.com/users/303604/articles/433788-drug-interaction- 11 Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. 12 Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. 13 Posted on Authorea 17 Mar 2020 — CC BY 4.0 — https://doi.org/10.22541/au.158447847.70587378 — This a preprint and has not been peer reviewed. Data may be preliminary. 14