pyrrolotriazines as novel potent alk inhibitors

Gregory J. Wells, Ph.D.

2,7-Disubstituted-Pyrrolo[2,1-f][1,2,4]triazines

N

NN

N

H

R'

R


A New Class Anaplastic Lymphoma Kinase (ALK)Inhibitors with in-vivo Anti-tumor Efficacy

Wells

Wells


General

Kinome - the set of protein kinases in an organism’s genome.

Kinases - enzymes that catalyze phosphorylation (from ATP) of amino acids, which fall into several groups and families:

• ones that phosphorylate serine and threonine;• ones that phosphorylate tyrosine;• some that phosphorylate both (eg. MAP2K and GSK families).

Essential for control and regulation of most biochemical pathways

• 518 known human protein kinases• 155 crystal structures solved• 160+ associated with human diseases• 14 small molecule inhibitors approved since 2001

Among the most active areas of targeted drug discovery in the past decade


ALK is a member of the TKL branch of the Protein Kinase Phylogenetic Tree

(Reprinted with permission from Cell Signaling Technology, Inc.)


ALK – Background

Anaplastic Lymphoma Kinase (ALK) is a cell membrane-spanning receptor tyrosine kinase with MW = 180 kDa

Most abundantly expressed in neonatal and adult brains, suggesting a possible role in brain and CNS development (Oncogene, 2001, 20, 5623)

Member of the insulin receptor (IR) family with IGF-1R, IRR, LTK and c-Ros

Binds to insulin receptor substrate 1 (IRS1); activates MAPK signaling pathway by activating the Ras-extracellular signal regulated kinase (ERK) pathway


ALK is implicated in the progression of certain tumors; approximately 60-70% of anaplastic large cell lymphomas (ALCL) are associated with a chromosome mutation containing a classical t (2;5) or other translocation that generates a mutant oncogenic fusion protein (NPM-ALK) consisting of nucleophosmin (NPM) and the intracellular domain of ALK

NPM-ALK possesses a constitutively active tyrosine kinase domain responsible for its oncogenic property through activation of downstream effectors

ALCL is a subset of high-grade non-Hodgkin’s lymphomas (NHLs) that typically presents as an aggressive systemic disease with ~50% 5-year survival rate after chemotherapy; predominant in children and young adults with about 2,500 new cases per year in US

Aberrant expression of constitutively active ALK is directly implicated in the pathogenesis of ALCL and ALK inhibition can markedly impair the growth of ALK+ lymphoma cells

Additionally, the EML4–ALK fusion gene has been identified in non-small-cell lung cancer (NSCLC) patients (Nature, 2007, 561) and represents another in a list of ALK fusion proteins that are promising targets for ALK inhibitor therapy.

Relation to Anaplastic Large-Cell Lymphoma (ALCL)


OCl

F

Cl

NNH2

N

N

NH

NH

CN

O

N

N

O

PfizerChugai Pharma.(Crizotinib)

N

NCl

NHNH

S

O

OO

N

N

N

Novartis (NVP-TAE684)

ALK IC50 = 3 nM

ALK IC50 = 2 nM

Competitor ALK Inhibitors

NH

O

NH

O

O

O

N

N

N

N

NH

NH

NH

N

O NHO

N

OF

GlaxoSmithKline

ALK IC50 = 0.5 nM

Chembridge

ALK IC50 = 0.2 MALK IC50 = 2 nM


Previously reported Cephalon ALK inhibitors

Wan, Cheng, Blood, 1617 (2006)ALK IC50 = 10nM

Karpas Cell IC50 = 150nM

Milkiewicz, Bioorg. Med. Chem., 4351 (2010)

N

N

N

NH

NH

N O O

ALK IC50 = 2nMKarpas Cell IC50 = 20nM

N NH

N

O

N

N F

F

N

NCl

NH

NH

O

N

N

O

O

N

O

N

NCl

NH

NH

NO

O CONH2


Mesaros, Bioorg. Med. Chem., 463 (2011)


Ott, Med. Chem. Lett., 493 (2010)


1 10 100 10000

30

60

90

120Sup-M2

CEP-14513 (nM)

Rel

ativ

e li

vin

g c

ells

p-NPM-ALK

NPM-ALK

― 30 100 300 (nM)

p-NPM-ALK

NPM-ALK

1 10 100 10000

30

60

90

120Karpas-299

CEP-14513 (nM)

Rel

ativ

e li

vin

g c

ells

1 10 100 10000

30

60

90

120 K562 cells

CEP-14513 (nM)

Rel

ativ

e li

vin

g c

ells

ALK Activity Is Essential for theProliferation of ALK+ ALCL Cells in Culture

NH

N

N

NO

O

NH

O(Me)2N

― 30 100 300 (nM)

CEP-14513

ALK IC50

Enzyme 3.7 nMCellular 10-30 nM


Conceptualization of Pyrrolotriazines as Potential ALK Inhibitors

N

NN

NHN

NCl

NHNH


Precedents in the Medicinal Chemistry Literature

O

OHOH

OH NNH

N

SO

OHOH

OH NN

N

NH2

N

NN

NHOH

N

NN

O

OH

NH

F

N

NN

O

F

NNH2

NH

NH

O

OF

NNH

N

NH

NH

N

NH

ON F

Purine/Adenosine mimics as C-linked nucleosides

Heterocycles, 569 (1992) Tet. Lett., 5339 (1994)

Quinazoline mimics as kinase inhibitors

JMC, 4059 (2004) JMC, 2143 (2006)

VEGFR-2 (BMS)

BMCL, 1945 (2007)

JMC, 7360 (2009)

IGF-1R (BMS)

c-MET (BMS)

Meijo University

Substituted Aminopyrazole hinge-binding motif

Albert Einstein College of Medicine


Crystal Structure of NVP-TAE684 bound to ALK

N

NCl

NHNH

O S

O

O

N

N

N

NVP-TAE684


Docking of a 2,7-pyrrolo[2,1-f][1,2,4]triazine analog with ALK

N

NN

NH

O

N

N

O


Synthesis of a Key Intermediate

2-Methylthio-7-bromo-pyrrolo[2,1-f][1,2,4]triazine

NH

O

OEt

N

O

OEt

NH2

N

O

OEt

NH

S

NH

O NH

NH

N

O

S

N

NN

Cl

S

N

NN

Cl

S

Br

N

NN

Cl

S

Br

N

NN

Cl

S

Br

Br

N

NN

S

Br

N

NN

S

Br

N

NN

S

Br

Br

+ +

+ +

Chloroamine

KO-t-Bu, THF

Benzoylthioisocyanate

THF

2N NaOH

1. MeI, THF

2. POCl3

NBS

THF- MeOH (2/1)

1. NaBH4, IPA, 55oC

2. DDQ, DCM

1 2 34

5 6 7 8

910 11


N

NN

S

Br

N

NN

S R2

N

NN

NH R2

R1

N

NN

S

O

N SO2Me

B(OH)2

R2

NH2

R1

Pd(OAc)2, Ph3P1. m-CPBA, DCM

2.

W, DIPEA, NMP

(R2 = OMe, -NHSO2Me)

9 12 13

14

1. m-CPBA, DCM

2. K2CO3, MeI

Targets from 2,7-Orthogonally Functionalized Intermediate

Method A


N

NN

S

Br

N

NN

NH

BrR1

N

NN

NH

R1

R2B(OH)2

R2

MeO OMe

OMe N

O

N

N

N

O

N

N

O

1. m-CPBA, DCM

2. R1-NH2, mW

DIEPA, NMP

Pd(OAc)2, PH3P

R1 =

9 15 16

Targets from 2,7-Orthogonally Functionalized Intermediate

Method B


Cpd. R1 ALK IC50 (nM)a IR IC50 (nM)a

17 212 ± 70 295 ± 95

18 >2000 194 ± 45

19 48 ± 8 35 ± 3

20 38 ± 11 19 ± 6

a IC50 values ± SD reported as the average of ≥ 3 determinations

OMe

MeO

MeO

N

O

N

N

N

O

N

NN

NH

R12

SAR of the 2-Position with 7-Phenyl-pyrrolo[2,1-f][1,2,4]triazine


SAR of the 2-Aryl-phenyl Group at C7

Cpd. R2 ALK IC50 (nM)a IR IC50 (nM)a

21 -OMe 20 ± 2 123 ± 37

22 -C(O)NH2 726 ± 113 326 ± 102

23 -SO2CH3 883 ± 233 > 2000

24 -NHSO2CH3 260 ± 57 1874 ± 485

25 -N(Me)SO2CH3 7 ± 2 366 ± 143

a IC50 values ± SD reported as the average of ≥ 3 determinations

N

NN

NH R2

N

N

7


In Vitro Profiles of Early Best Analogs

N

NN

NH

N

N

OMe

N

NN

NH N

N

N

SO2CH3

Liver microsome, t1/2(min)

(Mouse, Rat, Human)

CYP inhibition IC50 (M)a

3A4

KINOMEscan®

S(90) @ 1 mM

F (%)

(Rat)

8, 14, 40 9.4 0.524 4

30, 21, 40 14.6 0.614 --

a CYPs (1A2, 2C9, 2C19, 2D6): IC50 all > 30 M

21

25

Desired: Improved kinase selectivity and oral bioavailability


Eg. R1 R2

ALK

IC50 (nM)a

Cell

IC50 (nM)b

IR

IC50 (nM)a

KINOMEScan®

S(90) @ 1 M

F(%)

(Rat)

26 -OCH 9 ± 3 100 391 ± 80 0.085 --

27 -OCH 5 ± 21 100 206 ± 49 0.067 8

28 -OCH 4 ± 1 80 160 ± 36 0.067 6

29 -OCH 9 ± 3 85 357 ± 95 0.050 34

30 -OCH 9 ± 1 70 266 ± 65 0.164 38

N

NN

NH R2

O

R1

27

N

N

N

N

OH

N

N

OH

N

N

O

N

N

N

Optimized 2,7-Disubstituted Analogs

a IC50 values ± SD reported as the average of ≥ 3 determinationsb IC50 values reported as a mean of at least two determinations


Eg. R1 R2

ALK

IC50 (nM)a

Cell

IC50 (nM)b

IR

IC50 (nM)a

KINOMEScan®

S(90) @ 1 M

F(%)

(Rat)

31 -N(Me)SO2Me 9 ± 2 200 934 ± 323 0.187 --

32 -N(Me)SO2Me 10 ± 2 60 1137 ± 398 0.187 38

33 -N(Me)SO2Me 15 ± 5 80 1171 ± 169 0.194 41

34 -N(Me)SO2Me 7 ± 2 50 809 ± 299 0.187 47

N

NN

NH R2

O

R1

27

Optimized 2,7-Disubstituted Analogs

N

N

N

N

OH

N

N

OH

N

N

O

a IC50 values ± SD reported as the average of ≥ 3 determinations; b IC50 values reported as a mean of at least two determinations;c Kinase selectivity was determined using the Ambit Bioscience KINOMEscan® technology, and is expressed as S(90), the fraction of kinases inhibited >90% when screened at 1 μM across a panel of 256 kinases.


Anti-tumor Efficacy Comparison of Best Analogs

N

NN

NH

N

N SO2CH3O

N

O

N

NN

NH N

N

N

SO2CH3

OH

O

N

NN

NH

N

OMe

O

N

O 10mg/kg, po, bid 30 mg/kg, po, bid

(%TGI)

55 mg/kg, po, bid

(%TGI)

No significant effect 60 98

35% TGI 81 98

No significant effect 50 96

a TGI = tumor growth inhibition; calculated from tumor volume on final day

relative to vehicle control group

29

32

34


MTS Assay of 32 in ALK-positive and ALK-negative cell lines

Compound Conc. (nM)

N

NN

NH N

N

N

SO2CH3

OH

O

ALK positive lines

ALK negative lines


In-vivo Efficacy of 32 on Tumor Volumein ALK positive SUP-M2 Xenografts in Scid Mice

N

NN

NH N

N

N

SO2CH3

OH

O


N

NN

NH N

N

N

SO2CH3

OH

O

Plasma and Tumor Levels of 32 in ALKPositive SUP-M2 Xenografts in Scid Mice


N

NN

NH N

N

N

SO2CH3

OH

O

Pharmacokinetics of Lead Compound 32


Single Oral Dose PK/PD for Lead Compound 32

N

NN

NH N

N

N

SO2CH3

OH

O


t1/2 > 40 min (Mouse, Rat, Human liver microsomes)

Caco-2 permeability assay: Papp (AB) = 20.3 x 10-6 cm/s; PDR < 2

Plasma protein binding: Mouse (91%); Rat (68%); Humans (83%)

hERG (patch clamp): IC50 = 9.1 μM

CYP (1A2, 2C9, 2C19, 2D6, 3A): IC50 >12 μM

In vitro ADME and Safety Pharmacology Profile

N

NN

NH N

N

N

SO2CH3

OH

O

Compound nominated for pre-developmentand late stage pre-clinical safety studies

Ott, Wells; JMC 6328 (2011)


(MW = 514)

N

NN

NH

N

OMe

O

N

O

29

Unusual Mass Spectral Characteristics of Pyrrolo[2,1-f][1,2,4]triazines

- Infused alone - Infused w/ascorbic acid

Apparent formation of semiquinonediimine radical and quinonediimine ions (m/z = 514, 513, resp.) supported by their suppression with ascorbic acid

Evidence to suspect (potentially toxic) reactive metabolite formation

Similar observations with related pyrrolotriazine during metabolic studies


Proposed Mechanism of Ion Formation during ES-MS of Compound 29


UV Chromatograms from Microsomal Incubation of 32 containing GSH

(0 min)

(60 min)

N

NN

NH N

N

N

SO2CH3

OH

O


LC/MS Spectra of GSH Adduct of 32 Formed During Microsomal Incubation

Proposed fragmentation pattern

Wells-Knecht, Wells; Chem. Res. Toxicol., 1994 (2011)


Bioactivation/glutathione (GSH) trapping experiments in liver microsomes with a subset of analogs identified apparent NADPH-mediated oxidation products of reactive and potentially toxic quinone diimines.

Conflicting ADME vs. Analytical Trapping Experiments

Lead compound 32 showed good F, TGI, LMS, and acceptable safety pharmacology profile in several animal models.

However…

Prompting…

At this stage…

Further structural modifications on the suspected sites of oxidation to attenuatepotential formation of toxic metabolites to produce a “cleaner” drug candidate.


N

NN

NH N

N

N

SO2CH3

OH

O

I ncorporate EWGs on phenyl ringto lower oxidation potential

Replace heterocycle with piperidine to removethe p-nitrogen and lower the oxidation potential

Shif t piperazine ring f rom para to metato avoid f ormation of p-iminoquinone

I ncorporate small groups at C5to block potentially reactive site

Strategies to Mitigate Bioactivation/Glutathione Formation


Syntheses of Fluorophenyl- and m-Piperazine-Containing Analogs

X

R2

R4

R5

NO2

N

R2

R4

R5

NO2

NH

N

R2

R4

R5

NO2

NOH

N

R2

R4

R5

NH2

NOH

N

N

NH2

O

N

NN

OH R1

N

NN

NH R1

R2

R3

R4

R5

N

NN

R6 R1+

+

a b c

d or ee

3536 37 38

3940 41 42

aReagents and conditions: (a) (i) for X = F (inhibitors 11 and 13), N-Boc-piperazine, K2CO3, DMF, 60 °C, 60−90%, for X = Br (inhibitor 12), N-Boc-piperazine, Pd2(dba)3, XantPhos, Cs2CO3, 1,4-dioxane, 100 °C, 66%; (ii) TFA, DCM, rt, 80−97%; (b) (S)-2-methyl-oxirane, MeOH, 42−82%; (c) H2, Pd/C, MeOH, 87−97%; (d) 9a (R1 = N(Me)SO2Me), DIPEA, 1-methoxy-2-propanol, microwave, 200 °C, 8%; (e) (i) 9b, (F3CSO2)2NPh, DIPEA, DMF, 0 °C → rt; (ii) 8 or 10, 85 °C, 30−47%.


F- and m-Substitution on Aminophenyl Group Reduced GSH Adduct FormationR2

R3

R4

R5

NH

N

N

NR1

N

N

OH N

NA: B:

IC50 (nM)a

Cpd R1 R2 R3 R4 R5 ALK enzyme

ALK

cell

%GSH adductsb

32 -N(Me)SO2Me H A H -OMe 10 ± 2 60 61

43 -N(Me)SO2Me H A F -OMe 56 ± 16 -- 12

44 -N(Me)SO2Me F A H -OMe 14 ± 3 200 13

45 -N(Me)SO2Me F A H F 91 ± 19 -- 0.8

46 -N(Me)SO2Me B H H -OMe 28 ± 8 -- 3.9c

47 -OMe B H H -OMe 187 ± 64 -- 2.3c

aIC50 values reported as the average of at least two separate determinations; standard deviations are indicated where at least three determinations were made. bRelative to the tested compound. cDue to O-demethylation and iminoquinone formation/trapping


Nt-Boc

B

O

O

Cl

NO2

O

Nt-Boc

NO2

O

NR7

NH2

O

Nt-Boc

NH2

O

N

NN

R

ArNH

O

NR7

N

NN

R

ArNH2

N

NN

NH

O

N

O

O

R8

+

+

a b, c, d

e, b

or

f, e

e

g, b when R, R7 = H and Ar = o-methoxyphenyl

Synthesis of Piperidine Analogs

aReagents and conditions: (a) Pd(PPh3)4, KHCO3, water/1,4-dioxane, 80 °C, 97%; (b) TFA, DCM, rt, 95%; (c) for inhibitors 22, 23, 25, 27, 28, epoxide, MeOH, 42−80%; for inhibitors 31−40, BrCH2CONHMe or ICH2CONH2, Cs2CO3, MeCN, 75 °C, 71−80%; (d) H2, Pd/C, MeOH, > 90%; (e) (i) 20a, (F3CSO2)2NPh, DIPEA, DMF, 0 °C →rt; (ii) aniline (e.g., 19 or 21), 85 °C, 13−58%; (f) LiAlH4, THF, reflux, 92%; (g) N-Boc-aminoacid, EDCI, HOBt, DIPEA, DMF, 50−60%.


N

NN

NH

O

NR7

R1

ALK IC50 (nM)a Liver microsome t1/2 (min)b

Cmpd R1 R7 Enzyme Cellular M R Mo H

48 -N(Me)SO2Me 6 ± 1 60 > 40 21 < 5 < 5

49 -N(Me)SO2Me 6 ± 2 70 > 40 11 < 5 < 5

50 -N(Me)SO2Me H 10 ± 4 40 > 40 22 < 5 7

51 -OMe 3.4 ± 0.9 100 > 40 > 40 21 > 4

OH

OH

OH

Piperidine Analogs – Potency and Liver Microsome Stability

a IC50 values reported as the average of at least two separate determinations; standard deviations are indicated where at least three determinations were made. b Relative to the tested compound.


IC50 (nM)a Rat PK iv data

Cmpd R7 Ar ALK

Enzyme

ALK

Cellular

IR

Enzyme

S(90)b T1/2 (h) CL

(mL/min/kg)

Rat F%

52 Me- 6 ± 2 70 222 ± 84 0.10 1.4 29 3

53 3 ± 1 80 149 ± 55 0.09 1.4 62 16

54

3 ± 1 30 164 ± 42 0.06 2 39 15

55

7 ± 2 150 579 ± 194 0.09 2.6 25 13

56

11 ± 5 200 335 ± 87 -- 2.5 32 15

57

10 ± 4 100 161 ± 47 0.04 -- -- --

58

6 ± 2 100 222 ± 81 0.03 3.4 12 43

OH

F

OH

OH

NH2

O

O

NH

O

NH

O

NH2

O

O

O

O

O

O

O

a IC50 values are reported as the average of at least two separate determinations; standard deviations are indicated where at least three determinations were made. b Kinase selectivity was determined using the Ambit Bioscience KINOMEscan technology, and is expressed as S(90), the fraction of kinases inhibited >90% when screened at 1 μM across a panel of 256 kinases.

N

NN

NH

O

NR7

Ar

Piperidine Analogs – Potencies, Selectivity, and Rat PK


IC50 (nM)a Rat PK iv data

Cmpd R7 Ar ALK

Enzyme

ALK

Cellular

IR

Enzyme

S(90)b T1/2 (h) CL

(mL/min/kg)

Rat F%

58 6 ± 2 100 222 ± 81 0.03 3.4 12 43

59 12 ± 4 500 2967 ± 184 -- -- -- --

60 19 ± 6 250 747 ± 274 -- -- -- --

61 1060 ± 460 -- > 10,000 -- -- -- --

62 9 ± 4 150 410 ± 141 0.06 -- -- --

63 15 ± 4 80 552 ± 62 0.13 -- -- --

O

NH2O

aIC50 values are reported as the average of at least two separate determinations; standard deviations are indicated where at least three determinations were made. bKinase selectivity was determined using the Ambit Bioscience KINOMEscan technology, and is expressed as S(90), the fraction of kinases inhibited >90% when screened at 1 μM across a panel of 256 kinases.

N

NN

NH

O

NR7

Ar

Piperidine Analogs – Potencies, Selectivity, and Rat PK (cont.)

O

NH2

O

NH2

O

NH2

O

NH2

O

NH2

O

O

CN

N

O

N

O

N S

O

O


N

NN

NH

O

NNH2

OR

N SO

O

IC50 (nM)a Liver microsome t1/2 (min)b

Cmpd R ALK enzyme ALK Cell M R Mo H Rat F%

64 H 15 ± 4 80 29 < 5 < 5 <5 --

65 OH 11 ± 1 -- < 5 < 5 < 5 < 5 --

66 Me 6 ± 2 70 > 40 31 6 18 24

67 Cl 5 ± 1 70 > 40 > 40 15 37 41

IC50 values are reported as the average of at least two separate determinations; standard deviations are indicated where at least three determinations were made. bM = mouse; R = rat; Mo = monkey; H = human.

Piperidine Analogs – Potency and Rat PK for 5-Substituted Analogs

5

Mesaros, Thieu, Wells; JMC, 2012, 115


Pharmacokinetic Parameters of 58 in S-D Rats and Scid Mice

N

NN

NH

O

NNH2

O

O


PK/PD for 58 in Sup-M2 Xenografts in Scid Mice

N

NN

NH

O

NNH2

O

O

Inhibition of NPM-ALK, single dose 30mg/kg sol's in PEG400

Compound levels in plasma and tumor


Antitumor Efficacy of 58 in Scid Mice

N

NN

NH

O

NNH2

O

O


N

NN

NH

N

N

N

NN

NH N

N

N

SO2CH3

OH

O

N

NN

NH O

N

O

NH2

O

Pharmacological Milestones in the Development of Pyrrolotriazines as ALK Inhibitors

Potency SelectivityEfficacy

(TGI)Metabolic

Stability

? ?


Acknowledgements

OncologyBiologyChemistry

Henry BreslinJames DieboldArup GhoseDiane GingrichRobert HudkinsJoseph LiskoRobert McHughEugen Mesaros Karen MilkiewiczTho Thieu

DDS/Analytical

Hong ChangMangeng ChengLihui LuMatt QuailWendy WanAshley WohlerBruce Ruggeri

Lisa AimoneMark AlbomThelma AngelesLaura GwinnBeverly Holskin Zeqi HuangKristen Murray Damaris Steele Kelli ZeiglerSherri Meyer Mark Ator Joe Herman

Deborah LucianiKevin Wells-KnechtMehran Yazdanian

Jay TheroffGregory OttJonathan ParrishRabi TripathyTed UnderinerJason WagnerLinda WeinbergGregory WellsCraig ZificsakBruce Dorsey


Supplementary slides


Orton, Biochem J. 2005, v.392(Pt. 2), 249.

ERK Pathway


MAPK/ERK Pathway (alternate view)


Comparison of Crystal Structure of NVP-TAE684 andDocking of a 2,7-pyrrolo[2,1-f][1,2,4]triazine analog

N

NCl

NHNH

O S

O

O

N

N

N

NVP-TAE684

N

NN

NH

O

N

N

O


ALK IC50 (nM)CEP R1 R2 Enzyme Cell

11719 iBu HOCH2CC- 1.1 10 to 30

11834 iBu HON=C- 3.1 10 to 30

14083 nPr Ph(Me)NC(O)NH- 1.6 10 to 30

14513 iBu 3.7 10 to 30

11673 CH2cPr 428 >3000

11988 Et iBuON=C(Me)- 21838 >30000

ONH

OMe2N

O

Structure-Activity of Select Fused Pyrrolocarbazole Derivatives

NH

N

N

NR2

R1

O


1 10 100 10000

30

60

90

120 Sudhl-1

CEP-14513 (nM)

Rel

ativ

e li

vin

g c

ells

1 10 100 10000

30

60

90

120Sup-M2

CEP-14513 (nM)

Rel

ativ

e li

vin

g c

ells

― 30 100 300 (nM)

p-NPM-ALK

NPM-ALK

― 30 100 300 (nM)

p-NPM-ALK

NPM-ALK

― 30 100 300 (nM)

p-NPM-ALK

NPM-ALK

1 10 100 10000

30

60

90

120Karpas-299

CEP-14513 (nM)

Rel

ativ

e li

vin

g c

ells

1 10 100 10000

30

60

90

120 K562 cells

CEP-14513 (nM)

Rel

ativ

e li

vin

g c

ells

ALK Activity Is Essential for theProliferation of ALK+ ALCL Cells in Culture

NH

N

N

NO

O

NH

O(Me)2N

pyrrolotriazines as novel potent alk inhibitors

Documents

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n s n s ns n2

nm alk ic50

s ns n s n snbr br

n rnn nhrgregory

thf nbs nn n

active alk

theproliferation of