Heterocyclic Chemistry in Kinase Targeted Drug Discovery
Kazuya Kano, Yoshiaki Washio, Yasushi Miyazaki, Yutaka Maeda, Hideyuki Sato, Masato Nakano,
Masaichi Hasegawa Naohiko Nishigaki, Jun Tang
GlaxoSmithKline K. K., Tsukuba Research Laboratories, Chemistry Department
Introduction
Where the Selectivity of this Series
Comes from?
キナーゼ: ATPを介して基質であるセリン／スレオニンもしくはチロシ
ンの水酸基をリン酸化する酵素のことである。
Phe 80
Regulates: activity, location, degradation, conformation.
Many implicated in disease - especially cancer...
Kinase Systems Based Research to identify a
molecule for a kinase （我々はSystems Based Researchと呼
ばれる手法に基づいて創薬研究を行っている。この手法は従来の疾患由
来の創薬研究とは異なる）
•Start with a target family, not disease
•Allows us to apply what we learn from one member of the
class to others of interest
•More efficient - not a fresh start with each new target
•Build and use a knowledge base
Why it works
•ATP site - conserved but not optimized for ATP
•Large database of structures allows for:
•greater understanding of key pharmacophores and SAR
•improved homology models
•novel template design
Lys 85
CDK 2
Gatekeeper
GSK-3
N
N H
Br
HN
Lysine area
H2N
Biphenylureas, called “induced fit” moiety, at 5-position
greatly improve potency against angiogenesis related
kinases such as VEGFR’s, Tie2 and EphB4. GW 832757X
shows great Cellular activity(4.5nM)
Phe 80
CDK 2
Gatekeeper
Leu 132
As Angiogenesis Inhibitor
Leu 132
H
N
Imported Data From Chemically Aware Spreadsheet
O
Lysine area
Lys 85
CF3
Lead
Example 2
H
N
H2N
H
N
6.57
5.23
N
H
4.96
6.81
6.39
HN
8.94
8.63
8.57
F
N
H
CF3
8.63
Ar
N
R
Gate Keeper
N
H
N
N
Gate Keeper
induced fit
O
N
Cl
N
H
N
H
N
HN
Pan kinases <= 6.1
induced fit
inner
hydrophobic
OH Lys
N
Glu
H
H O
H
N H
Scaffold
N
Lys area
O
O
O
O H
N
Asp
Outer
R
Hydrophobic
O
N
N
H
O
N
O
H
N H
inner
hydrophobic
N
H
Scaffold
O N
Outer
H
Hydrophobic
Solvent Front
GW 850576X
Rock1 pIC50 = 7.2
JNK3 pIC50 = 6.9
VEGFR2, C-Fms pIC50 < 6.0
O
Lys
N
Glu
H
H O
H
Lys area
O
O
O
R
HN
Sugar Pocket
N
O
Asp
OH
Sugar Pocket
Solvent Front
In other cases
X-ray structure of ‘697X with c-fms
In C-Fms
GSK3 Inhibitor Design-starting from Tie2
legacy-Example 4
S
O
GW 671819X
GSK3 pIC50 = 7.1
CDK2 pIC50 = 4.7
H
N
HN
N
N
O
S
NH2
O
N
H
GW 822547X
CDK2 pIC50 = 9.5
Super selectiv e CDK2 inhibitor
Val
135
N
Pro
N
H
H
Tyr NN
NNH
O
H N
Lys
N
O
NH2
N
O
O HO
O
O
HN Asp
N
GW-671819X
SB-591475-A
N
H
N
H
O
O
N
SB-591475-A and GW-671819X docked into GSK-3
SB-591475A
GSK3 pIC50 = 7.5
GW 833547X
CDK2 pIC50 = 9.5
CDK1 pIC50 = 6.3
N
N
O
Strategy of GSK3 Inhibitor
Pursuance
New Template Development-Example 3
GW 680665X
N
F
Br
Core Structure
C-FMS
VEGFR2
TrJNK3
GSK3
6.23
6.10
5.82
5.87
OH
Gate Keeper
pIC50
induced fit
VEGFR2
src
others
N
inner
hydrophobic
OH Lys
N
Glu
H
H O
H
N H
Scaffold
N
Lys area
O
O
O
O H
N
Asp
Outer
R
Hydrophobic
N
O H
Gate Keeper
induced fit
inner
hydrophobic
Lys
N
Glu
H O
H
Lys area O
O
O
H
O
N
Outer
Hydrophobic
Sugar Pocket
H
N
136
NH2
New Template Development-Example 1
H
N
Asp
NH2
Overlap of GW 818954X and GW 833547X
H
N
133
O
O
N
HN N
N
H
N
GW 828604X
GSK3 pIC50 = 8.3
L6 glycogen Accumulatioin
Cellular Assay pEC50 = 6.4
O
N
GW 818954X
CDK2 pIC50 = 8.6
YAK1 pIC50 = 8.2
CDK1 pIC50 = 7.9
Publication
Example 1
H2N
N
N
O
H
S N
O
N
H
O
O
N
20 Months
O
NH2
N
N
N
H
GW 847320X
CDK2 pIC50 = 9
Cellular Assay IC50 = 3-5 uM
Lead
Example 4
NH
N
H
S
O
CF3
O
N
HN
N
H2N
GW 832757X
VEGFR2 pIC50 = 8.5
TIE2 pIC50 = 8.9
50 nM Cellular Activ ity(Huv ec V)
TRL Chemists
SBR(5410)
N
F
O
N
N
4.59
N
Lead
Example 3
Scaffold
1
7.3
In the case of c-fms, isoquinoline
demonstrates unique SAR caused by FLIP of
the template. This binding mode is supported
by x-ray structure analysis.
O
H2N S O
GW 847320X
CDK2 pIC50 = 9.0
Yak1 pIC50 = 8.3
HN
7.21
As C-FMS Inhibitor
Aminopyrazole versus Pyrrolopyrimidine
Similarity and Difference
H
HN
N
TRL Examples of Successful Kinase
Inhibitor Heterocycles
N H
O
H
N
H
N
SB 736764
c-Fms pIC50 = 7.67
VEGFR2 pIC50 = 6.72
7
O
GW
832757X
F
N
6.4
CF3
Pyrrolopyrimidine Hit Series-Example 2
HN
5
EPHB4
F
H
N
H
NH 2
O
VEGFR
3
N
CF3
H
N
Lys 33
Based on our research, some new information have been added to kinase
system including the concept of Lysine Area and how to design a selective
CDK-2 inhibitor.
H
N
N
H
GW
792416X
GW
824374X
O
H
N
HN
VEGF
R2
TIE-2
F
R
H2N H N
2
Lys 33
N
O
O
H
N
S
Ar
PIC50
Ar
GW
824372X
N
H2N
09 October 2002
R
GSK-3
H
N
5.5
5.5
<4.8
GW 586002X
The best in class
N
H
O
2
4
H NH
N
HO
HN
Attach acyl- to improv e
the interaction w ith FLP
and profile
Cut off 3-position
substituent
Solvent Front
O
N
N
O
N
Optimization of 2-position
substituent
Front lipophilic
pocket
Sugar Pocket
Salt bridge
O
3
O
Asp
NH2
N
O
GW 828604X
GSK3 pIC50 = 8.33
CDK2 pIC50 = 6.34
Solvent Front
The Difference between this Hit
and Historical Inhibitor
O
O S
NH
H
N
N
O
NH
O
O S
NH2
Modeling Analysis Comparison
As ALK5 Inhibitor
TGFb-Alk5 program for renal fibrosis needs new
back-up series because of selectivity issue over Alk4.
Hit series shown here shows excellent selectivity
versus ALK4.
Before modification
N
After modification
1
H
N
N
R
NH
R
S
pIC50
ALK4 ALK5
Ar
R
Br
GW 278534X
GW 798644X
CDK-2(pic50): 8.2
GSK-3(pic50): 7.3
CDK-2(pic50): 7.6
GSK-3(pic50): 5.6
pIC50
ALK4 ALK5
Ar
HN
HN
N
NH
4.61
7.71
NH2
6.3
7.08
6.93
7.13
6.2
6.61
N
OH
OH
S
NH
4.61
7.28
NH2
N
HN
OH
Overlap of X-ray structures of GW 278534X and
GW 798644X in the same CDK-2 protein.
Ar
5
NH
4.61
CF3
7.15
H
Overlap of GW 671819X and SB-591475A
Overlap of GW 809653X and SB-591475A
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