pyrrolotriazines as novel potent alk inhibitors
DESCRIPTION
Discovery and development of 2,7-Disubstituted-Pyrrolo[2,1-f][1,2,4]triazines. A New Class of Anaplastic Lymphoma Kinase(ALK)*Inhibitors with in-vivo Anti-tumor EfficacyTRANSCRIPT
Gregory J. Wells, Ph.D.
2,7-Disubstituted-Pyrrolo[2,1-f][1,2,4]triazines
N
NN
N
H
R'
R
Gregory J. Wells, Ph.D.
A New Class Anaplastic Lymphoma Kinase (ALK)Inhibitors with in-vivo Anti-tumor Efficacy
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
ALK is a member of the TKL branch of the Protein Kinase Phylogenetic Tree
(Reprinted with permission from Cell Signaling Technology, Inc.)
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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)
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
ALK IC50 = 4nMKarpas Cell IC50 = 50nM
Mesaros, Bioorg. Med. Chem., 463 (2011)
ALK IC50 = 14nMKarpas Cell IC50 = 45nM
Ott, Med. Chem. Lett., 493 (2010)
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
Conceptualization of Pyrrolotriazines as Potential ALK Inhibitors
N
NN
NHN
NCl
NHNH
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
Crystal Structure of NVP-TAE684 bound to ALK
N
NCl
NHNH
O S
O
O
N
N
N
NVP-TAE684
Gregory J. Wells, Ph.D.
Docking of a 2,7-pyrrolo[2,1-f][1,2,4]triazine analog with ALK
N
NN
NH
O
N
N
O
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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.
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
N
NN
NH N
N
N
SO2CH3
OH
O
Plasma and Tumor Levels of 32 in ALKPositive SUP-M2 Xenografts in Scid Mice
Gregory J. Wells, Ph.D.
N
NN
NH N
N
N
SO2CH3
OH
O
Pharmacokinetics of Lead Compound 32
Gregory J. Wells, Ph.D.
Single Oral Dose PK/PD for Lead Compound 32
N
NN
NH N
N
N
SO2CH3
OH
O
Gregory J. Wells, Ph.D.
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)
Gregory J. Wells, Ph.D.
(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
Gregory J. Wells, Ph.D.
Proposed Mechanism of Ion Formation during ES-MS of Compound 29
Gregory J. Wells, Ph.D.
UV Chromatograms from Microsomal Incubation of 32 containing GSH
(0 min)
(60 min)
N
NN
NH N
N
N
SO2CH3
OH
O
Gregory J. Wells, Ph.D.
LC/MS Spectra of GSH Adduct of 32 Formed During Microsomal Incubation
Proposed fragmentation pattern
Wells-Knecht, Wells; Chem. Res. Toxicol., 1994 (2011)
Gregory J. Wells, Ph.D.
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.
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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%.
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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%.
Gregory J. Wells, Ph.D.
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.
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
Pharmacokinetic Parameters of 58 in S-D Rats and Scid Mice
N
NN
NH
O
NNH2
O
O
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
Antitumor Efficacy of 58 in Scid Mice
N
NN
NH
O
NNH2
O
O
Gregory J. Wells, Ph.D.
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
? ?
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
Supplementary slides
Gregory J. Wells, Ph.D.
Orton, Biochem J. 2005, v.392(Pt. 2), 249.
ERK Pathway
Gregory J. Wells, Ph.D.
MAPK/ERK Pathway (alternate view)
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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
Gregory J. Wells, Ph.D.
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