英-[药物报告]色瑞替尼

Ceritinib(Zykadia ®)Research code: LDK-3781 General Information● Ceritinib is a kinase inhibitor, which was first approved in2014 by US FDA.● Ceritinib was discovered and marketed by Novartis. ● Ceritinib inhibited autophosphorylation of ALK, ALK-me-diated phosphorylation of the downstream signaling protein STAT3, and proliferation of ALK-dependent cancer cells.● Indicated for the treatment of patients with anaplastic lym-phoma kinase (ALK)-positive metastatic non-small cell lung cancer (NSCLC) who have progressed on or are intol-erant to crizotinib.● Available as capsules with each containing 150 mg ofceritinib and recommended dose is 750 mg once daily until disease progression or unacceptable toxicity.Key Approvals around the World *First approvaldate 04/29/2014 06/05/2015 NDA NO. 205755 EMEA/H/C/003819Brand name Zykadia ® Zykadia ® Indication NSCLC NSCLCAuthorisation holderNovartisNovartis Europharm Ltd*Till Dec 2015, it has not been approved by PMDA (Japan) and CFDA (China).Worldwide SalesActive IngredientMolecular formula : C 28H 36N 5O 3ClS Molecular weight : 558.14 CAS No.: 1032900-25-6Chemical name : 5-Chloro-N 4-[2-[(1methylethyl)sul-fonyl]phenyl]-N 2-[5-methyl-2-(1-methylethoxy)-4-(4-pi-peridinyl)phenyl]-2,4-pyrimidinediamine Parameters of Lipinski's “Rule of 5”5583871144.70 ± 0.95aCalculated by ACD/Labs software V11.02.Drug Product *Dosage route : Oral Strength : 150 mg Dosage form : CapsuleInactive ingredient : Colloidal anhydrous silica, L-hydroxy-propylcellulose, magnesium stearate, microcrystalline cel-lulose, and sodium starch glycolate, gelatin, indiogotine, and titanium dioxide Recommended dose :The recommended starting dose is 750 mg once daily with-out food.Discontinue ceritinib for patients unable to tolerate 300 mg daily.If a dose of ceritinib is missed, make up that dose unless the next dose is due within 12 h.*Sourced from the FDA drug label information.2 ▐2014 Worldwide NCEsKey PatentsPatents Summary•Ceritinib’s compound patent application was filed as PCT application by IRM in 2007.•The compound patent will be expired in 2027 originally, which has been granted in Japan, Europe, the United States successively.•Zykadia® (Ceritinib) has got five years NCE market exclusivity protection and seven years ODE after it was approved by FDA in Apr. 29, 2014 initially.Patents ListCeritinib ▐ 32 ChemistryRoute 1: Original Discovery RouteSynthetic Route: The overall synthetic route of ceritinib was enabled via a modified Buchwald-Hartwig coupling of two advanced intermediates aryl amino-piperidine 6 and arylsulfonyl chloro-pyrimidine 7. Aryl amino-piperidine 6 was synthesized in five stepsfrom 2-chloro-4-fluorotoluene 1. Nitration of 1 using KNO3/H2SO4 yielded the corresponding nitro derivative 2 in a 71% yield. Condensation of compound 2 with iso propanol in the presence of Cs2CO3 provided the 2-chloro-4-iso propoxy-5-nitrotol-uene 3 in excellent yield. Suzuki coupling of compound 3 with 4-pyridineboronic acid 4 provided compound 5 in 73% yield. Concomitant reduction of pyridine and nitro group using PtO2/H2 afforded the corresponding piperidine-aniline intermediate, which was subsequently protected with Boc, generating compound 6 in 60% over two steps. Buchwald-Hartwig coupling of 6 with arylsulfonyl chloro-pyrimidine 7 provided the precursor of ceritinib. Boc deprotection with TFA, concentration, and subse-quent precipitation with 1 N HCl yielded ceritinib as the HCl salt in 35.0% yield from 6 and the overall yield of 10.3%.[1, 2] Synthesis of the required arylsulfonyl chloro-pyrimidine coupling precursor 7 began with 2-(isopropylsulfonyl)benzenamine 8 and 2,4,5-trichloropyrimidine 9 under basic condition (NaH, DMF, DMSO)in 60% yield.4 ▐2014 Worldwide NCEsRoute 2:Synthetic Route: In this strategy, ceritinib was prepared from reduction of 4-(5-isopropoxy-2-methyl-4-nitrophenyl)pyridine 1 under Pd/C H2 condition afforded corresponding phenylamine 2 in 92% yield. Then Sandmeyer reaction was carried out in the presence of NaNO2/CuBr/HBr/H2O to give 4-(4-bromo-5-isopropoxy-2-methylphenyl)piperidine 3 in 83% yield. Buckward-Hartwig coupling of 3 with 5-chloro-4-nitropyrimidin-2-amine 4 converted to desired compound 5 in 78% yield, which was hy-drogenated and coupled with 1-bromo-2-(isopropylsulfonyl)benzene 7 to obtain the target ceritinib in 74% yield over two steps. The overall yield was 44%.[3]3 PharmacologySummaryMechanism of Action●Ceritinib is a kinase inhibitor, which inhibited ALK, insulin-likegrowth factor 1 receptor (IGF-1R), insulin receptor (InsR), andROS1.●Ceritinib was approximately 50-fold more specific for ALK(IC50 = 0.15 nM) than InsR (IC50 = 7 nM) and IGF-1R (IC50 = 8nM), other members of theinsulin receptor superfamily.●Ceritinib inhibited autophosphorylation of ALK, ALK-mediatedphosphorylation of the downstream signaling protein STAT3,and proliferation of ALK-dependent cancer cells.●The off-target activity of ceritinib was evaluated for 42 of thetarget receptors (MC4, IC50 = 0.6 nM), 84 of the broad spectrumscreen receptors (transporter monoamine, inhibition = 97%), and10 of the GPCRs (dopamine 2 receptor, IC50 = 4000 nM).Ceritinib ▐ 5 In Vitro Efficacy●Anti-proliferative activity of ceritinib in tumor cell lines:Cell lines expressed with ALK fusion protein: IC50 = 11-56 nM.Cell lines expressed with EML4-ALK-mutation: IC50 = 37.6-250 nM.Ba/F3 cells expressed with others fusion proteins: IC50 = 180-400 nM.●Phosphorylation of ceritinib in Karpas299 cells:ALK protein: IC50 = 46 nM.STAT3 protein: IC50 = 150 nM.In Vivo Efficacy●H2228 cells xenograft models:In SCID mice: Significance at doses ≥6.25 mg/kg. Complete tumor regression with 25 mg/kg ceritinib after 14 days.In nude rats: Significance at doses ≥10 mg/kg.●Karpas299 cells xenograft models:In SCID mice: Significance at doses ≥12.5 mg/kg. Complete tumor regression with 25 mg/kg ceritinib after 14 days.In nude rats: Significance at doses ≥12.5 mg/kg. Complete tumor regression with 25 mg/kg ceritinib after 14 days.●Crizotinib-resistant H2228 cells xenograft model carrying the ALK-mutation in SCID mice:Non-ALK-mutation: Significant at doses ≥50 mg/kg.I1171T ALK-mutation: Significant at doses ≥25 mg/kg.C1156Y ALK-mutation: Significant at doses ≥50 mg/kg.Mechanism of ActionTable 1In Vitro Effects of Ceritinib and Crizotinib on Human Protein Kinases[4]EPK CE ALK (1066-1459) Y a0.15 3 EPK CE AXL Y 180 13EPK CE IGF-1R (980-1369) Y 8 400 EPK CE RET (658-1072) Y 400 2200 EPK CE INSR (871-1343) Y 7 290 EPK ROCK2 S/T 450 2500EPK AURORA_A S/T b110 600 EPK CE FGFR3(411-K650E-806) Y 430 1700EPK cABLT315 Y 130 6The applicant evaluated the selectivity of ceritinib and crizotinib by testing its in vitro activity against 36 recombinant human protein kinases using the Caliper mobility shift assay, table 1 showed ceritinib most selective for ALK (IC50 <0.5 μM). a Y: Tyrosine-specific protein kinases. b S/T: Serine/Threonine-specific protein kinases.[4]H2 receptor 170 NAmonoamineRabbit 10 μM 97 331Opk receptor 570 NA K channel[Ka] Rat 10 μM 99 682MC3 receptor 670 NA Somatostatin sst1 Human 10 μM 88 2420MC4 receptor 600 35% at 10 μM Somatostatin sst2 Human 10 μM 76 2250Ad3 receptor 730 8300 Somatostatin sst3 Human 10 μM 93 5120NK1 receptor 990 79% at 30 μM Somatostatin sst4 Human 10 μM 90 1880D2 receptor 1200 NA 5-HT5A Human 10 μM 71 5000The off-target activity of ceritinib was evaluated using an in vitro safety pharmacology profiling panel including 139 G protein-coupled receptors (GPCRs), ion channels, nuclear receptors, transporters, and enzymes that have been previously linked to potential side effects. At least eight concentrations of ceritinib were tested. The agonist and antagonist activities of ceritinib were evaluated for 10 GPCRs using a safety pharmacology screen. A fluorometric imaging plate reader (FLIPR) assay was conducted with a four-fold, eight-point serial dilution of ceritinib up to 10 μM to a ssess agonist and antagonist activity. Ceritinib showed weak agonist activity on the dopamine D2 receptor (EC50 = 6 μM, IC50 = 4 μM), but did not exhibit significant activity against the other GPCRs tested. a Ceritinib interacted with 42 of the target receptors, table 2 showed the target receptors IC50 <1500 nM. b A safety pharmacology broad spectrum screen with 84 receptors at ceritinib was tested at 10 μM in duplicate, table 2 showed the broad spectrum receptors inhibited >70%.6 ▐2014 Worldwide NCEsIn Vitro EfficacyTable 3 In Vitro Anti-proliferative and Phosphorylation of Ceritinib[4]Anti-proliferative activityBa/F3Murinepro-B lymphoma cell withALK fusion kinaseNPM-ALK 26-35NATEL-ALK-Q1 56EML4-ALK 27 Ba/F3Murinepro-B lymphoma cell withALK-mutationEML4-ALK I1171T 37.6 440EML4-ALK L1196M 69 1460EML4-ALK G1202R 94 1370EML4-ALK C1156 160 340EML4-ALK S1206A 160 270EML4-ALK G1269S 250 2140 Ba/F3Murinepro-B lymphoma cell withtyrosine kinaseTEL-ROS 180NATEL-IGF-1R 220TEL-InsR 400Karpas299 Human ALCL NPM-ALK 45 (n = 8)NA NCI-H2228 Human NSCLC EML4-ALK 11 (n = 5) NB-1 Human neuroblastoma cell ALK 24 (n = 3)Endogenous phosphorylation Karpas299 Human ALCLALK protein 46NASTAT3 protein 150The cell viability following 48 h of treatment with control compounds or ceritinib serially diluted with DMSO was determined using the Bright-Glo™ luciferase biolumi-nescent assay and cell proliferation was measured using the Brite Lite™ luciferase assay. ALCL: Anaplastic large-cell lymphoma. NSCLC: Non-small cell lung cancer. NA: Not available.In Vivo EfficacyTable 4 In Vivo Efficacy of Ceritinib in Different Xenograft Models[4]H2228 HumanNSCLCSCIDmouse(female)Wild type3.125 QD × 14416.25Complete tumor regressionwas observed even at adose of 25 mg/kg/day af-ter 14 days.6.25 QD × 143612.5 QD × 14-6425 QD × 14-100RNU nuderat(female)Wild type5 QD × 146710Complete tumor regressionwas observed even at adose of 25 mg/kg/day af-ter 14 days.10 QD × 14 225 QD × 14-100Ceritinib ▐ 7ContinuedC1156Y ALK-mu-tation25 QD × 12 61.8 50 Complete tumor regres-sion was observed even at a dose of 100 mg/kg/day.50 QD × 12 11.7 100 QD × 12 -100 Karpas299Human ALCLSCID mouse (female)Wild type6.25QD ×14 62 12.5 Complete tumor regres-sion was observed at 25 mg/kg after 14 mg/kg/day.12.5 QD × 14 18 25 QD × 14 -93 RNU nuderat (female)Wild type6.25QD × 14 74 12.5Significant tumor regres-sion was observed at 25 mg/kg/day after 14 days.12.5QD × 143025 QD × 14 -33 50QD × 14-66Study: H2228 NSCLC xenograft mice model Animal: SCID beige mice (female)Model: 5 × 106 NCI-H2228 cells were implanted subcutaneously into the right hind flank of one SCID mouse.Administration: Treated daily, p.o., ceritinib: 3.125, 6.25, 12.5 and 25 mg/kg/day, ve-hicle control: NAStarting treatment: Mice bearing established tumors (~85 mm 3) Test: Tumor volumes, 3 times a weekResult: Treatment with 25 mg/kg ceritinib resulted in complete tumor regression after 14 days. ** P <0.05.Figure A Effect of Ceritinib on Human H2228 NSCLC Xenografts in SCID Mice Model [4]8 ▐2014 Worldwide NCEsStudy: Crizotinib-resistant H2228 cells xenograft with ALK I1171T mutations micemodelAnimal: SCID beige mice (female)Model: Crizotinib-resistant H2228 NSCLC with ALK I1171T mutations were im-planted subcutaneously into one SCID mouse.Administration: Treated daily, p.o., ceritinib: 12.5, 25, or 50 mg/kg/day, crizotinib100 mg/kg/day, vehicle control: NAStarting treatment: Mice bearing established tumors (~130 mm3)Test: Tumor volumes, 3 times a weekResult: Treatment with 50 mg/kg ceritinib resulted in complete tumor regression after14 days of treatment. (**P≤0.05; ****P≤0.0001)Figure B Effect of Ceritinib on Crizotinib-resistant Human H2228 NSCLC Xenograft with ALK I1171TMutations in SCID Mice Model[4]Study: Karpas299 ACLC xenograft mice modelAnimal: SCID beige mice (female)Model: 5 × 106 Karpas299 cells were implanted subcutaneously into the right hindflank of one SCID mouse.Administration: Treated daily, p.o., ceritinib: 6.25, 12.5 and 25 mg/kg/day, vehiclecontrol: NAStarting treatment: Mice bearing established tumors (~74 mm3)Test: Tumor volumes, 3 times a weekResult: Treatment with 25 mg/kg ceritinib resulted in c significant tumor regressionafter 14 days of treatmentFigure C Effect of Ceritinib on Human Karpas299 ALCL Xenografts in SCID Mice Model[4]4 ADME & Drug-Drug InteractionSummaryAbsorption of Ceritinib●Exhibited a non-linear pharmacokinetics in humans after oral administration. The increase in AUC appeared to be greaterthan dose-proportional in the dose range of 50 to 750 mg ceritinib.●Had a moderate bioavailability in rats (48.3%), but high in mice (54.6%) and dogs (58%).●Was observed slowly (T max = 3.98-15 h) in humans, mice (7 h), rats (12 h) and monkeys (13-18.3 h).●Showed a half-life of 19.4-40.6 h in humans, much longer than those in rats (13.2 h) and monkeys (12.1-16 h), after oral ad-ministration.●Had a moderate clearance in mice (26.6 mL/min/kg), rats (1.49 L/h/kg), but low to moderate in monkeys (0.366-0.78 L/h/kg),after intravenous administration. The Cl/F in humans was 44.5-126 L/h after oral administration.●Exhibited an extensive distribution in mice, rats and dogs, with the apparent volume of distribution at 9.7, 19.9 and 6.53-13.5L/kg, after intravenous administration. The V z/F in humans was 1880-6230 L after oral administration.●Ceritinib was classified as a low passive permeability compound.Distribution of Ceritinib●Exhibited high plasma protein binding in rats (97.9-98.4%), dogs (98.3-98.4%), monkeys (94.4-97.4%) and humans (96.7-98.8%). Note that ceritinib was mainly bound to HSA.●The binding to RBC was 56.9-58.6% in humans, indicating the drug was distributed more to blood cells than to plasma.●In Long Evens rats after oral and intravenous administration:Ceritinib ▐ 9 The drug was widely distributed into most tissues including central nervous system although with a lower degree com-pared to other tissues.Relatively higher concentration levels were observed in intestinal wall, uveal tract, pituitary gland, bile, adrenal cortex, harderian gland, liver, spleen, lymph node, lungs, kidneys, thyroid, bone marrow, adrenal medulla, pancreas, thymus and salivary gland. The concentration in tissues was generally higher than blood concentration.The radioactivity concentration in the uveal tract was 176 times of that in blood, suggesting a high affinity for melanin-rich tissues.Ceritinib-derived radioactivity was retained in several other tissues (including testis, epididymis, and skin) and the elimi-nation was not yet complete by 168 hours post-dose.Metabolism of Ceritinib●Could slightly metabolized in rat, monkey and human hepatocytes.●CYP3A was the major metabolizing enzymes, with CYP2C19, 1A2, 2C8, 2D6 and 2C9 involved in the metabolism ofceritinib.●The metabolism of ceritinib included mono-oxygenation, O-dealkylation, S-dealkylation, and N-formylation of ceritinib, andsecondary di-oxygenation, glucuronidation, sulfation and dehydrogenation.●Overall, the parent drug was the most abundant component in plasma in humans. Eleven metabolites were found in theplasma of humans, each at levels ≤ 2.3% of the total drug-related AUC.●Five of these eleven metabolites were not detected in the plasma of either the rat or monkey. The remaining three uniquehuman metabolites detected at low levels in plasma included M46.6 (1.7%), M48.8 (1.7%), and M52.0 (2%).Excretion of Ceritinib●Was predominantly eliminated in feces in rats, monkeys and humans, with the parent drug as the significant component in rat,monkey and human feces.●In cannulated rats, excretion was mainly into the bile 65.4% after oral administration and 24.3% after intravenous administra-tion.Drug-Drug Interaction●Ceritinib was a strong inhibitor of CYP3A4/5 (IC50= 0.2 μM), and moderate of CYP2A6 (IC50= 5 μM), CYP2B6 (IC50 = 2μM), CYP2C8 (IC50= 25 μM), CYP2C9 (IC50= 2 μM), CYP2C19 (IC50= 70 μM), CYP2D6 (IC50 = 20 μM) and CYP2E1 (IC50= 30 μM).●Ceritinib had no induction for CYP1A2, CYP2B6 or CYP3A4 mRNA/activities, but had induction for CYP2C9 in vitro at theconcentration of 0.25-2.5 μM ceritinib.●Ceritinib was a substrate of P-gp and BCRP in vitro, but not inhibitors of them.●Ceritinib was not a substrate of OCT1, OAT2, OATP1B1 or OATP2B1.●Ceritinib inhibited OATP1B1 or OATP1B3, OAT1 and OCT2, with the IC50 of >5 μM in vitro, but did not inhibit OAT3 orOCT1.AbsorptionTable 5 In Vivo Pharmacokinetic Parameters of Ceritinib in Mice, Rats and Monkeys after a Single Dose Administration[4]Balb/c mouse (male) i.v. 5 -nM nM·h nM·h a6.2 ± 0.5mL/min/kg9.7 ± 0.6 - p.o. 20 7.0 ± 0695 ± 31nM12296 ± 981nM·h10334 ± 963nM·h a- - - 54.6 ± 4.4HanWistar rat (male) i.v. 10 0.083 ± 0 975 ± 139 6950 ± 1470 6890 ± 1510 9.7 ± 1.2 1.49 ± 0.342 19.9 ± 0.49 - p.o. 25 12.0 363 8390 8330 13.2 NA NA 48.3Cynomolgus monkey (male) i.v. 5 0.083 1410 6530 6450 29 0.78 13.5 -i.v. 10 0.083 3190 27800 27400 14.5 0.366 6.53 -p.o. 30 18.3 ± 9.81 881 ± 12.5 35800 ± 3460 35500 ± 3520 12.1 ± 2.05 - - 43.0 ± 4.17 p.o. 60 13 ± 9.2 947 ± 140 45300 ± 8860 45100 ± 8840 16 ± 0.61 - - 58a AUC0-24.10 ▐2014 Worldwide NCEsTable 6 In Vivo Pharmacokinetic Parameters of Ceritinib in Patients Following a Single Doses of Ceritinib on[5]50n = 1 n = 1 n = 1 n = 1 n = 1 n = 1 n = 1100 19.4n = 115.0(6.00-24.0)n = 229.3(10.1)n = 2467(10.7)n = 2938(24.3)n = 23250n = 1116n = 1200 33.2n = 15.08(4.17-6.00)n = 240.2(88.5)n = 2703(55.6)n = 21460(62.9)n = 23720n = 177.5n = 130030.1(10.0)n = 34.00(4.00-5.95)n = 3198(41.5)n = 33440(44.7)n = 37470(46.5)n = 31880(50.7)n = 244.5(36.8)n = 240030.7(39.1)n = 104.99(2.97-6.73)n = 12120(80.9)n = 121920(78)n = 124070(81.8)n = 123470(74.4)n = 595.9(58.6)n = 550031.1(11.1)n = 73.98(3.00-23.5)n = 8153(86.5)n = 82350(87.9)n = 85140(142)n = 86230(219)n = 3147(170)n = 360037.6(24.6)n = 66.00(3.00-24.1)n = 9212(59.7)n = 93590(53.4)n = 98180(57.4)n = 91990(4.30)n = 246.39.60)n = 270038.9(98.4)n = 36.00(4.00-25.0)n = 4206(146)n = 43450(138)n = 49210(112)n = 42340(56.5)n = 266.6(35.8)n = 275040.6(34.7)n = 96.02(3.95-23.8)n = 10186(127)n = 103390(121)n = 107870(127)n = 104230(164)n = 388.5(163)n = 3n: Number of patients with non-missing values. Values were median (range) for T max, geometric mean (CV% of geometric mean) for all others.Key finding:Ceritinib was classified as a low passive permeability compound.DistributionTable 7 In Vitro Plasma Protein Binding and Blood Partitioning of [14C]Ceritinib[5]Rat 97.9 ± 2 98.4 ± 0 98.4 ± 0.1 100 97.6 ± 0.6 Rat0.4 3.7 1.3Dog 98.8 ± 0.5 98.3 ± 2.1 98.4 ± 0.810000 98.6 ± 0.8 Dog66.6 ±1.667.0 ±0.471.6 ±1.1Monkey 94.4 ± 0.5 97.4 ± 0.4 95.2 ± 0.2 Monkey 78.7 ±1.177.8 ±1.576.4 ±0.4Human 98.8 ± 0.5 97.2 ± 1.4 96.7 ± 1.8 Human 58.2 ±3.558.6 ±5.556.9 ±4.8[4] FDA Datadase. /drugsatfda_docs/nda/2014/205755Orig1s000PharmR.pdf (accessed Nov 2015).14[4]Blood 63.4 600 220 48.0 NM NM 1.00 1420 1 Brain 29.8 42.0 20.0 13.0 NM NM 0.148 137 0.096 Bile NM 74000 29000 11000 NM NM 155 48800 34.4 Lungs 607 18000 5900 2100 28.9 21.2 37.2 23300 16.4 Liver 2720 33000 8100 1800 78.7 130 52.6 29500 20.8 Kidney cortex 693 17000 6400 1300 20.3 28.3 32.8 23700 16.7 Stomach glandular 13400 5200 1900 310 NM NM 26.3 6590 4.64 Adrenal cortex 1870 37000 11000 4100 41.4 NM 72.4 48700 34.4 Harderian gland 31.5 1700 1800 2600 1060 2990 62.8 5040 3.55 Bone marrow 198 **** **** 2700 23.3 NM 28.0 9220 6.50 Small intestine wall 164000 490000 11000 730 NM NM 413 6980 4.92 Thyroid gland 843 18000 7300 1200 NM NM 30.3 35100 24.7 Pituitary gland 229 14000 6600 15000 1810 2590 211 15000 10.6 Spleen 1040 27000 6400 2500 42.3 41.6 47.6 NA NA Lymph node 200 19000 2900 4100 183.0 NM 45.1 8720 6.14 Adrenal medulla 890 21000 2100 1300 48.1 NM 26.5 14600 10.3 Pancreas 388 13000 4000 1300 6.6 NM 24.6 14200 9.98 Thymus 120 4300 1400 2400 86.7 NM 21.3 3860 2.72 Salivary gland 208 9700 3500 1700 NM NM 20.2 8740 6.16 Uveal tract 108 6500 1100 8500 2880 8160 176 8600 6.06 NM: Not measured,Metabolism14[5]Rat 2.5 0-18 NC NC NCNC 12.5 0-24 NC NC NCMonkey 2.5 0-4 7.02 98.7 312.90.12 12.5 0-8 18.0 38.5 131.7Human 2.5 0.25-18 100.4 6.90 17.30.014 12.5 0-18 66.95 10.4 25.8NC: Not calculated.[4] FDA Datadase. /drugsatfda_docs/nda/2014/205755Orig1s000PharmR.pdf (accessed Nov 2015).14[6]Ketoconazole (3A) 0-1 0.100 ~0.03 90.5 Azamulin (3A) 0-5 0.150 ~0.01 100 Ticlopidine (2B6/2C19) 0-10 1.70 >10 10.9S-mephenytoin (2C19) 0-250 95.5 >250 18 Montelukast (2C8) 0-2 0.0140 (0.18-0.71) >2 16.5 Furafylline (1A2) 0-10 2.00 >10 19.1 Quinidine (2D6) 0-1 0.0605 >1 17.9 Sulfaphenazole (2C9) 0-5 0.510 >5 11.9 Fluvastatin (2C9) 0-5 0.525 >5 21.6a Median value were calculated from (DMPK R0400785).If a range of values was reported for a study an average value was taken. The K i or IC50 values used in the calculations corresponded to inhibition of the enzyme (or subfamily, in the case of CYP3A).b Percent maximal inhibition of total [14C]Ceritinib metabolism at the concentrations of inhibitor examined.Rat a Ceritinib 108.03 ± 2.34 324.73 ± 17.09 426.58 ± 62.86 609.30 ± 32.39 L746530 168.29 ± 1.3 460.89 ± 22.3 709.78 ± 43.0 1068.22 ± 17.5Human a Ceritinib 105.80 ± 7.02 281.74 ± 0.68 504.46 ± 16.86 702.25 ± 57.12 L746530 173.98 ± 8.8 523.11 ± 0.5 684.59 ± 70.4 1057.12 ± 76.3Rat b Ceritinib 11.59 ± 0.25 34.73 ± 1.83 45.62 ± 6.72 65.17 ± 3.46 L746530 18.00 ± 0.14 49.29 ± 2.38 75.91 ± 4.60 114.25 ± 1.88Human b Ceritinib 11.32 ± 0.75 30.13 ± 0.07 53.95 ± 1.80 75.11 ± 6.11 L746530 18.61 ± 0.94 55.95 ± 0.05 73.22 ± 7.53 113.06 ± 8.16a Based on viable cells.b Based on 107 × 106 hepatocytes per gram of tissue (Wilson, et at 2003). Values were mean ± rang (n = 2) and rounded to three significant figures or to the nearest 0.01.[6]CeritinibRat 10.0 ± 1.2 21.2 ± 1.3 21.4 ± 0.9 15.0 ± 1.9 Human 10.4 ± 1.3 20.5 ± 0.5 12.3 ± 10.7 11.3 ± 0.9L746530Rat 64.4 ± 1.7 715.9 ± 35.7 579.8 ± 35.0 590.0 ± 49.5 Human 52.8 ± 2.6 254.3 ± 5.2 245.0 ± 24.1 354.1 ± 33.8a Based on nominal microsomal protein included in incubation. Values were mean ± SD (n = 3). All values were rounded to three significant figures or to the nearest 0.1.Monkey 2.50.25 92.5 0.23 0.31 ND 3.33 ND 0.68 1.20 ND ND0.5 89.9 0.28 0.59 ND 4.68 ND 0.77 1.77 ND ND1 83.8 0.81 0.95 ND 8.67 ND 0.58 3.58 ND ND2 76.9 1.96 1.26 ND 10.6 1.00 5.15 ND ND ND4 69.1 2.37 2.13 2.20 13.4 1.46 7.15 ND ND ND8 61.5 2.97 3.01 4.27 14.8 2.69 8.12 ND ND ND18 50.6 4.06 2.40 5.11 20.4 2.68 8.21 ND 2.51 2.9124 32.0 7.11 4.42 11.5 25.0 5.23 8.77 ND 2.15 2.5 12.50.25 NA NA NA NA NA NA NA NA NA NA0.5 92.7 0.23 0.24 ND 1.95 0.20 0.37 1.92 ND ND1 89.3 0.34 0.56 ND 3.11 0.41 0.53 3.09 ND ND2 80.7 1.68 0.97 1.09 6.05 1.17 5.60 ND ND ND4 80.6 1.12 1.21 1.56 5.32 1.81 6.12 ND ND ND8 74.2 2.16 1.34 4.06 5.96 3.48 5.90 ND ND 0.9318 71.2 2.10 1.43 5.61 6.81 3.34 5.07 ND 0.89 1.6924 61.9 4.05 1.81 8.11 9.31 5.82 5.41 ND 0.87 1.40Human 2.5 0.25 90.4 - - - 0.78 - 0.75 1.76 - ND0.5 90.9 - - - 0.77 - 0.98 1.19 - ND1 91.8 - - - 1.28 - 1.35 1.14 - ND2 91.6 - - - 2.22 - 1.18 1.02 - ND 4 89.9 - - - 3.11 - 1.46 1.26 - ND 8 87.6 - - - 4.52 - 1.47 1.55 - 1.11 18 83.2 - - - 8.17 - 1.61 1.52 - 1.59 24 81.4 - - - 8.94 - 2.62 1.38 - 2.28Human 12.5 0.25 NA - - - ND - NA NA - NA0.5 96.4 - - - ND - ND 3.60 - ND1 95.5 - - - ND - ND 4.47 - ND2 95.5 - - - ND - ND 4.55 - ND 4 95.7 - - - ND - ND 4.26 - ND 8 95.2 - - - ND - ND 4.81 - ND 18 85.1 - - - 7.05 - ND 7.89 - ND 24 90.8 - - - 2.96 - 3.93 0.67 - 1.65-: Not observed.Table 14 In Vivo Metabolites Profiles in Plasma, Bile and Feces of Rats, Monkeys and Humans[6]Plasma aRat i.v./p.o. 10/25 100 - - - - - - - - - - Monkeyi.v. 10 89.9 1.4 - 1.8 0.6 1.2 - 3 ---p.o. 30 84.4 3.6 - 2.5 0.4 1.6 - 3.1 ---Human p.o. 750 mg 81.6 - 1.2 1.4 1.9 1.6 1.8 2.3 1.7 1.7 2Bile Rat i.v. 10 34.89 ND ND - 1.09 3.26 0.83 ----p.o. 25 9.19 1.71 0.84 - 0.72 1.46 0.52 ----FecesRati.v. 10 12.05 3.75 4.56 - ND ND ND ----p.o. 25 51.77 4.54 5.88 - ND 2.82 ND ----Monkeyi.v. 10 55.1 1.8 2.8 - 3.7 10.9 17.9 0.3 ---p.o. 30 60.2 2.7 2.2 - 2.7 5.9 8.7 0.4 ---Human p.o. 750 mg 68 - 3.9 - 1.8 1.4 6.5 1.6 1.1 1.4 2.2M0: Ceritinib. a %AUC for metabolites in plasma.Figure D Proposed Pathways for In Vivo Biotransformation of [14C]Ceritinib in Rats, Monkeys and Humans[6] The red labels represent the major components in matrices. P = Plasma, U = Urine, B = Bile, F = Feces.ExcretionRat (intact)10 i.v. 0-168 - 0.241 ± 0.174 107 ± 7.825 p.o. 0-168 - 0.180 101 Rat (BDC)10 i.v. 0-72/168 65.4 0.62 29.825 p.o. 0-72/168 24.3 ± 8.52 1.05 ± 1.03 65.0 ± 17.0 Monkey10 i.v. 0-168 - 0.59 10530 p.o. 0-168 - 0.71 ± 0.45 92.3 ± 6.94 Human 9.14 p.o. 0-168 - 1.2 91.0Drug-Drug Interaction[5, 6]CYP1A2 Phenacetin >100 (2.5) - 1.0CYP1A2 mRNA 1.01 1.14 1.07 32.1CYP2A6 Coumarin 5 (1.5) 0.03 (0.009) 61.0 Activity 1.15d 1.22 1.01 31.7CYP2B6 Bupropion 2 (0.3) 5.3 (0.780) 1.3CYP2B6 mRNA 1.02 1.17 0.96 10.2CYP2C8 Paclitaxel 25 (0.6) - 1.1 Activity 1.05 1.20 0.92 4.65CYP2C8 Amodiaquine 2 (0.6) 16.7 (4.86) 1.1CYP2C9 mRNA 1.12 1.15 1.18 2.34CYP2C9 Diclofenac 2 (0.6) 0.24 (0.0701) 8.5 Activity 1.27 1.77 1.10 3.50CYP2C19 S-mephenytoin 70 (1.8) - 1.0CYP3A4 mRNA 1.27 2.73 6.03b31.1CYP2D6 Bufuralol 20 (2.9) - 1.2 Activity 0.98 1.43 1.33c8.81 CYP2E1 Chlorzoxazone 30 (4.4) - 1.1CYP3A4/5 Midazolam 0.2 (0.06) 0.16 (0.0469) 12.3CYP3A4/5 Testosterone 0.2 (0.06) -19.0Calculation of R1 values based on maximal steady-state concentration of 1010 ng/mL or 1.8 μM. [I]/K i assumed to be IC50/2 for competitive inhibition for those CYP enzymes without an experimental K i value. a Omeprazole 50 μM for CYP1A2, phenobarbital 1000 μM for CYP2B6 and CYP2C9, rifampin 10 μM for CYP3A4. b Mean of 3.24-, 8.74-, and 6.12-fold change relative to vehicle control in the three human hepatocyte lots. c Mean of 0.936-, 0.983-, 2.06-fold change relative to vehicle control in the three human hepatocyte lots. d:Mean of 1.75-, 1.24-, and 2.31-fold change relative vehicle control in the three human hepatocyte lots.[5]。