CHIR-258, a Novel Multi-targeted
Tyrosine Kinase Inhibitor, for the
Treatment of t(4;14) Multiple Myeloma
Suzanne Trudel
Princess Margaret Hospital
Toronto, Canada
Chromosomal Translocations in Myeloma
The identification of IgH translocations in MM has resulted in the development of
powerful prognostic tools and identification of novel molecular targets
Five recurrent loci are most frequently involved
46.3 mo
other
Patients with a t(4;14)
18 mo
translocation have poor
t(4;14)
prognosis despite high dose
chemotherapy
OS in months from the time of transplant
Chang H. et al, Br J Heamatol. 125: 64; 2004
t(4;14) MM Expresses FGFR3
a
FGFR3-FITC
t(4;14) Negative
t(4;14) Positive
Chang H., et al, Blood 2005
Activating FGFR3 Mutations are Present in
Some t(4;14)Tumors
G384D
K650E
S249C
K650M
R248C
Y373C
J807C
I
II
III
TM
TK1
TK2
S - S
S - S
S - S
*
*
Model for the Multi-step Pathogenesis of t(4;14)
Myeloma
PC
MGUS
MM
PCL
chromosomal translocation
additional genetic alterations
activating
t(4;14)
mutations
Ectopic expression of
FGFR3 and MMSET
FGFR3 or N, K-ras
MAPK activation
proliferation
Increased bcl-xL
anti-apoptosis
Inhibition of FGFR3 Kinase Activity Prevents
Malignant Transformation
Trudel et. al, Blood 103: 3521; 2004
CHIR-258 is a Potent Inhibitor of Class III, IV & V
Receptor Tyrosine Kinases
I
II
III
IV
V
F
N
N
NN
N
NO
CHIR-258
IC50 in uM
RTK
FLT3
<0.001
c-KIT
0.002
CSFR1/c-fms
0.036
FGFR1
0.008
FGFR3
0.009
VEGFR1/Flt1
0.01
VEGFR2/Flk1
0.013
VEGFR3/Flt4
0.008
PDGFR
0.027
IGFR1
PDGFR
0.21
INSR
2
EGFR1
2
c-MET
>3
EphA2
4
CHIR-258 IC >2 µM
50
CHIR-258 IC
< 210 nM
50
TIE2
4
IGFR1
>10
HER2
>20
Adapted from Grassot et al. Nucleic Acids Res. 2003 Jan 1; 31(1): 353-8.
CHIR-258 Inhibits FGFR3 Signaling and Viability of
FGFR3 Expressing Human Myeloma Cell Lines
Table 2. IC50 values (in nM) of CHIR-258 aga inst human myeloma cell lines
Cell line
t(4;14)
FGFR3
IC50 (nM)
genotype
KMS11
+
Y373C
90
KMS18
+
G384D
550
OPM2
+
K650E
90
H929
+
WT
>2500
8226
-
N/D
> 2500
U266
-
N/D
>2500
Unstimulated
DMSO
100 nM CHIR-258
FGF
500 nM CHIR-258
Cell Cycle Pathway Genes Downregulated by
CHIR-258 in KMS11 Cells
Values are a mean of two experiments
Untreated Control
Treated for 48 hours (500nM)
CCND3
CCND3
CCND2
CCND2
CHIR-258 Induces Apoptosis of FGFR3
Expressing HMCLs
CHIR-258 Inhibits
Viability of KMS11
Cells in the Presence of
IL-6, IGF-1 and Bone
Marrow Stroma Cells
0
100 nM
CHIR-258
500 nM
Response Surface Analysis of CHIR-258 and
Dexamethasone on a KMS11 Cells Indicates a Synergistic
Interaction
Devi
CHIR-258ation
(left from
to
Addi
right)
ti
x vity
dex -
( Data
back to front)
dex x dex control
Deviation from Additivity - Data
additive
synergism
log [dex]
additive
log [dex]
antagonism
Deviation
CHIR-258 xfrom Additivity - Da
CHIR-258 control
ta
log [258]
-8.0
-7.0
-6.0
-6.0
-6.5
-7.0
-7.5
-8.0
log [dex]
additive
· Observe synergy of CHIR-258 from and dexamethasone, following
log [258]
simultaneous addition at 72 hour after administration.
log [258]
· Dose ranges of both drugs are physiologically relevant.
CHIR-258 Inhibits KMS11 Myeloma Growth
Real-time monitoring in vivo using bioluminescence imaging
X.Xin, Chiron Corp.
CHIR-258 Demonstrates Increased Survival Benefit
against KMS11-Luc Orthotopic Model
100
80
* 5/9 mice remaining
60
Survival 40
1. Vehicle
%
2. CHIR-258 at 20 mg/kg/d
20
started at day 2
0
0
10
20
30
40
50
60
70
80
90 100
Time (days)
* Log rank test: CHIR-258 vs. Vehicle; p=0.0001
X.Xin, Chiron Corp.
CHIR-258 Inhibits Proliferation and
Induces Apoptosis of FGFR3
Expressing Myeloma Tumors In vivo
4G10
FGFR3
FGFR3
0
10
30
60 mg/kg
Placebo
H & E
KI-67
Cleaved Caspase 3
CHIR-258
60mg/kg
CHIR-258 Induces Apoptosis of FGFR3 Expressing
Primary Myeloma Cells
Unstimulated
Isotype control
ts
FGF/DMSO
Anti-FGFR3
nuo
Counts
FGF/500 nM CHIR-258
C
FGFR3
pERK
DMSO
CHIR-258
Summary: 4/5 FGFR3 +ve patients responded; 0/5 in FGFR3 -ve group
% apoptosis responders: DMSO 9.2% vs CHIR-258 24.5% (p=0.049)
Summary
· CHIR-258 is a novel, multi-targeted RTK inhibitor with potent
activity against FGFR3 and a target profile (VEGFR, CSFR1) that
suggests the potential to favorably modulate the bone marrow milieu
· CHIR-258 inhibits both WT and mutant FGFR3 and downstream
signaling via the MAP kinase pathway
· CHIR-258 has cytotoxic activity against FGFR3 expressing human
myeloma cell lines and primary patient samples
· IL-6, IGF-1 and co-culture on stroma does not confer significant
resistance to CHIR-258
· CHIR-258 therapeutic efficacy was demonstrated in a xenograft
model of FGFR3 MM
· In conclusion, the studies support the clinical evaluation of CHIR-258
for these poor prognosis patients
Acknowledgements
Princess Margaret Hospital
CHIRON Corporation
Zhi Hua Li
Marion Weismann
Ellen Wei
Katherine Rendahl
Keith Stewart
Eddie Moler
Donna Reece
Carla Heise
Hong Chang
Xiaohua Xin
Christine Chen
Joseph Mikhael
MMRF, ASH Scholar Award, Canadian Institute Health Research