1. No category

Preclinical activity in non-Hodgkin’s lymphoma of selinexor, a Selective Inhibitor of Nuclear Export (SINE),
is enhanced through combination with standard-of-care therapies
Asfar S. Azmi1, Amro Aboukameel1, Robert O. Carlson2, Sivan Elloul2, Sharon Shacham2, Michael Kauffman2, Ran Frenkel3, Ramzi M. Mohammad1
1Wayne State University, Detroit, MI; 2Karyopharm Therapeutics, Newton, MA; 3Karyopharm Europe GmbH, Munich, Germany
ABSTRACT
RESULTS
RESULTS
DLBCL cell
line
Selinexor
IC50 (µM)
RL
0.020 0.050 0.057 0.063 0.070 0.096 0.120 0.150
0.29
0.44
0.48
Control
SEL
EVER
SEL+EVER
DLBCL cell lines were incubated with a range of selinexor concentra8ons over 72 hr. Resul8ng cell viability was determined using an MTT-­‐based assay. (*Double hit)
1200
B
Vehicle
CHOP
KPT-276 (75 mg/kg)
1000
KPT-276 (150 mg/kg)
KPT 251 (25 mg/kg)
KPT 251 (75 mg/kg)
800
avg. tumor vol. day 1: 125 ± 58 mm3
P<0.05 vs vehicle (Mann Whitney):
CHOP Day 10 and 12; KPT-276 (150 mpk) Day 12
600
400
200
0
2
4
6
8
10
12
15
C
Vehicle
vehicle
rituximab 20 mg/kg IV
selinexor 20 mg/kg PO
selinexor 30 mg/kg PO
CHOP
KPT-276 (75 mg/kg)
KPT-276 (150 mg/kg)
10
KPT 251 (25 mg/kg)
75
KPT 251 (75 mg/kg)
5
0
**
50
****
**
25
avg. mouse et Day 1: 20.2 ± 1.5 g
-5
**p<0.01; ****p<0.0001 vs vehicle (Greenwood)
P<0.05 vs vehicle (Mann Whitney):
all treatments, all time points Day 3-12,
except KPT-276 (75 mpk) on Days 3 and 12
-10
Time (days)
0
2
4
6
8
10
12
0
Time (days)
0
10
20
30
40
50
60
70
80
90
Time (days post cell inoculation)
Equivalent in vivo efficacy of single agent selinexor vs rituximab or CHOP. [A] and [B]
Xenograft model of DLBCL. SINE Compounds KPT-251 and KPT-276 were administered
sc and po, respectively, in cycles of once daily for ten consecutive days with a one day break
prior to start of a new cycle. Cyclophosphamide, doxorubicin and vincristine (CHO) was
administered once IV at MTD and prednisone (P) was administered po QDX5. [C] 10X106
WSU-FSCCL follicular lymphoma cells were injected IV in the tail veins of ICR-SCID mice
and vehicle or drug treatments were started one week later.
C vs Dex = 0.451209899
C vs Kpt = 0.383789488
C vs Combo = 0.025863099
this is for day 25 using ttest on excel, tail=2, type=2
Selinexor synergizes with either DEX or EVER. WSU-FSCCL or WSU-DLCL2 cells were
incubated with 100 nM selinexor (SEL) or 100 nM dexamethasone (DEX) or 1.25 μM
everolimus (EVER), each drug alone, SEL+DEX or SEL+EVER. [Left Panel] Resulting cell
viability was determined using trypan blue staining and cell counting. [Right Panel] Annexin
V FITC apoptosis analysis (Sel 100 nM; DEX 100 nM and EV 2.5 μM). SEL (100 nM)
- + - +
- + - +
SEL (100 nM)
- + - +
- + - +
EVER (2.5 µM)
- - + +
- - + +
DEX (100 nM)
- - + +
- - + +
WSU-DLCL2
PARP
PARP
Caspase 3
Caspase 3
XPO1
XPO1
β-actin
β-actin
WSU-DLCL2
WSU-FSCCL
WSU-FSCCL
SEL+Dexamethasone
SEL+Everolimus
2000
Control
SEL PO 10 mg/kg
Ever PO 2.5 mg/kg
SEL+Ever (10+2.5)
1500
1000
500
0
750
500
250
0
7
18
21
25
28
32
Days Post WSU-DLCL2 Bilateral Trasplantation
Control
SEL PO (10 mg/kg)
DEX IP (7.5 mg/kg)
SEL+DEX (10+7.5)
1000
7
18
21
25
WSU-DLCL2 days post transplantation
Selinexor at sub-optimal doses enhances the activity of DEX or EVER in subcutaneous
DLBCL xenografts. WSU-DLCL2 xenograft were established as described above. Drugs
were administered at indicated doses 5 days a week for three weeks. [Left panel] SEL
+EVER and [Right Panel] SEL+DEX combination (study continuing beyond 25 days).
CONCLUSIONS
KPT-­‐251
KPT-­‐276
DB
0.55
A
100
Percent survival
SEL+DEX
median mouse wt (% change from day 1)
DEX
0
selinexor
OCILY A3/ OCILY SUDH SUDH DOHH WSUD SUDH TOLE PFEIF
3
KAW
19
L5
L8
2*
LCL2
L6
DO
FER
SEL
Tumor Weight (mg)
Methods: Diffuse large B cell lymphoma (WSU-DLCL2) and follicular small cleaved cell
lymphoma cell lines have been previously developed at Wayne State University. All lines are
GCB type with the exception of OCI-LY3 (ABC), WSU-DLCL2 (neither) and A3/KAW
(unknown). Cell growth inhibition was performed using trypan blue viability assay and MTT
assay (CellTiter 96® from Promega) and IC50 values were calculated using GraphPad Prism®
software. Apoptosis was detected using Annexin V FITC assay. Changes in protein
expression was evaluated using western blotting. For xenograft model of DLBCL, pieces
(~50 mg) of serially passaged WSU-DLCL2 tumors were transplanted into the flanks of 4-5
wk old ICR-SCID mice and vehicle or drug treatments were started one week later. 10X106
WSU-FSCCL follicular lymphoma cells were injected IV in the tail veins of ICR-SCID mice
and vehicle or drug treatments were started one week later. Control
Tumor Weight (mg)
METHODS
WSU-FSCCL disseminated follicular lymphoma model
WSU-DLCL2 DLBCL xenograft model
median tumor volume (% of day 1)
Introduction: The nuclear export protein Exportin 1 (XPO1) is
overexpressed in diffuse large B-cell lymphoma (DLBCL), follicular
small cell cleaved lymphoma (FSCCL) and a wide variety of other
cancers, which often correlates with poor prognosis. Selinexor is an oral
SINE currently in Phase 1/2 clinical testing, which targets XPO1 to
induce apoptosis across a broad spectrum of tumor types. This broad
action is primarily due to forced nuclear retention and reactivation of
tumor suppressor proteins (TSPs), resulting in selective tumor cell death.
Here we report combination studies involving selinexor/dexamethasone
and selinexor/mTOR inhibitor everolimus in Non Hodgkin’s lymphoma
relative to front-line standard-of-care therapies. RESULTS
Molecular analysis of SEL-DEX and SEL-EVER synergy. WSU-DLCL2 or WSU-FSCCL
were exposed to indicated concentrations of drugs for 72 hrs followed by protein isolation
and western blotting. [Upper panel] Results showing enhanced PARP cleavage, full length
caspase 3 reduction and decrease in XPO1 expression for combination treatments compared
to single agents. β-actin was used as loading control. [Lower panel] Structures of SINEs.
•  The enhanced in vitro and in vivo efficacy of selinexor in combination with DEX or the
mTOR inhibitor everolimus provides rationale for further clinical investigation.
•  The equivalent efficacy of single agent selinexor vs rituximab or CHOP is consistent
with observed clinical activity of selinexor in refractory DLBCL and follicular
lymphoma.
•  A Phase 2 study of selinexor in DLBCL is currently recruiting patients. (Study of
Selinexor (KPT-330) in Patients With Relapsed/Refractory Diffuse Large B-Cell
Lymphoma” (SADAL – NCT02227251).