ANTIMYELOMA EFFICACY OF PLITIDEPSIN (APLIDIN®):
FROM BENCH TO THE BEDSIDE
Enrique M. Ocio1,2, Constantine Mitsiades3, M. Victoria Mateos1, Patricia Maiso2, Faustino Mollinedo2, Mercedes Garayoa2, Consuelo Gajate2, Joan Bladé4, Felipe Prosper5, Juan José Lahuerta6, Nicholas Mitsiades3,
Ciaran J. McMullan3, Nikhil C. Munshi3, Teru Hideshima3, Dharminder Chauhan3, Carmen Cuevas7, Pablo Avilés7, Glynn Faircloth7, Paul G. Richardson3, Atanasio Pandiella2, Kenneth C. Anderson3, Jesus F. San Miguel1,2
1Hospital Universitario de Salamanca, Spain; 2Centro de Investigación del Cįncer. Universidad de Salamanca, Spain; 3Jerome Lipper Multiple Myeloma Center, Dana-Farber Cancer Institute, USA 4Hospital Clinic de
Barcelona, Spain; 5Clķnica Universitaria de Navarra, Spain; 6Hospital 12 de Octubre, Madrid, Spain; 7PharmaMar SAU, Clinical R&D, Colmenar Viejo-Madrid, Spain.
BACKGROUND
MATERIAL & METHODS
· The outcome of MM patients has improved in the recent years with the introduction of novel therapeutic agents.
· In vitro studies were performed in 23 multiple myeloma (MM) cell lines and in cells from 16 MM patients.
Nevertheless most patients are refractory or relapse after achieving a response. Therefore, the development of new
compounds is still needed to overcome the drug resistance displayed by myelomatous plasma cells.
· For the in vivo analysis a human plasmocytoma model in CB17-SCID mouse was used. Mice were randomized to
receive Aplidin® 100 µg/Kg ip x 7 days/week (n=9), Aplidin® 140 µg/Kg ip x 5 days/week (n=7) or vehicle alone (n=9).
· Plitidepsin is a cyclic depsipeptide isolated from the marine tunicate, Aplidium lbicans with promising antitumor activity.
· The clinical efficacy of Aplidin® in relapsed/refractory patients was evaluated in a Phase II trial. Dosage of Aplidin® was
· This work represents a comprehensive study (in vitro, in vivo and clinical) of its antimyeloma efficacy.
5 mg/m2 every 2 weeks. Dex was added after 3 cycles if PD or after 4 cycles if SD.
RESULTS
In vitro efficacy
Mechanism of action
Clinical trial
Aplidin® is very effective against MM cell lines
Aplidin® overcomes the
Treatment of MM1S with Aplidin® induce
35 relapsed/refractory MM patients have been included. Median number of previous
Aplidin (nM)
proliferative effect of IL-6
caspase dependent apoptosis
lines of therapy: 4 (1-9). 12 pts have received Thal-based combinations and 15 pts
0
0
1
5
10
20
50 100
5
MM1S
L363
l)
MM1R
and
IGF-1
but
only
Bortezomib-based regimens.
10
U266
tro
U266
120
n
U266LR7
LR5
l)
15
ation
100
co
partially that of BMSCs
Time with Aplidin 100 nM, hours
INA-6
tro
20
80
n
ive
MR20
25
ct
60
co
e
0
1
3
6
12
24
kDa
e
etaboliz
RESPONSE
N (%)
NCI-H929
30
40
iv
m
20
·Data from 29 evaluable patients for APL alone
160
PARP
112
ARD
35
T
Dose response of
T
fresp
Cleaved PARP
85
Partial abrogation of
ect
o
0
140
ARK
p
40
M
0
0.1
1
10
100
MM1S
(%
l) 120
Caspase 3
o
35
take
tr
MM1S + IL-6
S6B45
45
[Aplidin], nM
MM1S by Annexin V
p
n 100
Cleaved
Aplidin®-induced apoptosis
fres
u
of
OPM-1
o
50
U
80
rd
o
fragments
·Best Response for APL alone:
%
B
(%
60
19
JJN3
55
17
with a pan-caspase inhibitor
40
KMS-18
ty(
60
Caspase 7
35
ili
20
100
IC50 1-10 nM
RPMI-8226/S
ls
PR
2 (6.9%)
ab
65
0
Cleaved
0
M
M
M
M
M
M
K620
70
n
1 n
5 n
n
n
n
80
10 nM 50
cel
fragments
·2 patients (6.9%) obtained a PR
llvi
10
50
100
ib
ib
LP-1
e
+
20
75
om
V
60
C
Aplidin Aplidin Aplidin Aplidin
100
Dox40
Aplidin
n
80
Caspase 8
57
ls
BortezomBortez
xi
40
e
43
OCI-MY5
85
cel
80
ls
Granulo-Monocytes
n
100
n
20
Cleaved
+
OPM-6
160
90
Aplidin 100 nM
·3 patients (10.3%) obtained a MR
cel
A
V
60
80
140
0
fragments
n
ARP-1
+
%
Z-VAD-FMK
95
V
MM1S
MR
4 (13.7%)
60
10
xi
l) 120
0
1
3
6
10
24
40
e
Z-IETD-FMK
100
in
MM1S + IGF-I
40
take
tro 100
47
Caspase-9
n
p
n
u
on
Time with Aplidin 100 nM (h)
37
20
Z-LEHD-FMK
20
80
nnex
U
35
A
A
rd
of
0
60
%
0
%
B
(%
Bcl-2
26
·9 patients (31.0%) obtained a SD
40
Control
Ap 10 nM
Ap 100 nM
0
6
12
Time (hours)
tient 4
tient 5
tient 9
30
20
Bcl-X
Pa
Pa
Pa
tient 13
Pa
Efficacy against cells
0
Mcl-1
40
0
M
M
nM
nM
M
M
1 n
5 n
n
n
10
50
Plasma cells
Lymphocytes
Myc
63
100
10 nM 50
PR+MR
6 (20,6%)
ls
ls
ib
ib
100
100
·Best Response for APL/Dex (5 pts):
lidin
lidin
lidin
from 16 MM patients
Ap
Aplidin
om
cel
cel
Ap
Ap
80
80
Aplidin
+
+
BortezomBortez
V 60
V 60
in
in
40
40
160
without toxicity in
Aplidin® induces CD95/Fas
nnex 20
nnex 20
140
MM1S
·
1 pt in SD ---
SD
A
0
A
0
) 120
%
%
MM1S + BMSC
ake
trol 100
normal cells
tient 4
tient 9
upt
on
translocation into lipid rafts
tient 4
tient 5
tient 9
tient 13
tient 5
tient 13
f 80
Aplidin®-induced apoptosis is, at
Pa
Pa
Pa
SD
12 ( 41 ,3%)
Pa
Pa
Pa
Pa
Pa
rdU
o 60
B
(%
·
4 pts in PD --
1 pt MR
40
% of AnV+ PC
in MM144 cells
least partially, mediated through
20
Patient Previous treatments
0
Ap 10 nM
Ap 100 nM
0
M
M
nM
M
nM
3 pts SD
5 n
n
10 n
50
100
50
JNK and p38 activation
ib
1
At diagnosis
100
100
Control
Aplidin 10 nM x 15h
lidin
lidin
Aplidin
om
Ap
Ap
AplidinBortez
2
At diagnosis
78
100
1 2 3 4 5 6 7 8 9 10 11 12
1 2 3 4 5 6 7 8 9 10 11 12
3
VBCMP/VBAD+ASCT; Bortezomib; TACYDEX
96
99
GM-1 cont.
lipid rafts
p-c-jun
IP: -JNK
APL single agent
APL/ Dex
4
MP; VBAD; Bortezomib
83
97
5
At diagnosis
53
92
Aplidin® potentiates the effect
CD95/Fas
p-p38
-p-p38
6
At diagnosis
69
89
tive
p-Erk1/2
-p-Erk1/2
Overall
Over
Median TTP
2.2 mo
m . (1.5
(1
- 2.5)
2.4 mo . (1.6
(1. - 3.1)
7
VBCMP/VBAD
33
83
of other antimyeloma agents
Erk1/2
-Erk1/2
8
VBCMP/VBAD+ASCTx2
52
81
Sensi
9
At diagnosis
32
74
Erk5
IP: -Erk5
Median TTP in PR/MR
5.4 mo
m . (1.9
(1
- 7.6)
7.9 mo . (1.9
(1. - 8.6)
120
-Erk5
10
VBCMP/VBAD+ASCT; High Dex; TACYDEX
44
71
Aplidin® induces deregulation
ek ) 100
PARP
-PARP
11
VBCMP/VBAD+ASCT;Bortezomib
40
66
tap
80
Median TTP in SD
2.7 mo. (2.3
(2. - 4.7)
3.1 mo . (2.3 - 4.7)
C
15'
3h
6h
12h
24h
C
15'
3h
6h
12h 24h
of 805 genes by GEP analysis
12
MD; VBAD
37
64
u
ontrolc 60
bitol
30' 1h
9h
bitol
30' 1h
9h
Sor
Sor
t
TT
13
At diagnosis
39
61
(%
40
Aplidin 100 nM
Bortezomib 25 nM
n
M
20
Analysis of those involved in
14
MP
7
33
sta
0
Aplidin
- +
- +
- +
- +
- +
- +
15
At diagnosis
0
22
1.25 nM
apoptosis and cell cycle
Dexam
Bortez
Melph
Lenal
Thal
Toxicity profile
Resi
16
At diagnosis
2
18
12.5 nM 1.25 nM 1.25 µM 1 µM 200 µM
Cell cycle
Apoptosis
Partial abrogation of Aplidin®-induced
Average
Average
80
Grade 1-2
Grade 3-4
Fold Change
Fold Change
apoptosis with the pretreatment of MM1S
74
-2.7
39.0
-2.9
25.9
71
-3.1
15.2
cells with a JNK- and a p38 kinase -inhibitor
In vivo efficacy
-3.2
10.1
ents
-3.2
9.2
-3.4
9.2
60
ati
-3.4
7.2
-3.5
7.0
SP600125
SB203580
P
7.0
1800
Vehicle vs treated
-3.8
47
-3.8
6.0
Aplidin® reduces tumor
Vehicle
1.0
p=0.008
-3.8
4.4
of
41
Apl 100 µg/Kg x 7d/w
-4.0
4.0
3.6
1400
-4.2
%
Apl 140 µg/Kg x 5d/w
100
Control
40
-4.2
3.5
growth and increases
(mm3)
0.8
ls
32
al
-4.4
3.4
SB203580
v
2.9
80
me
-4.6
cel
SP600125
2.9
27
27
lu
1000
rvi
0.6
+
u
Log Rank
-4.6
-2.9
60
V
26,5
survival in a human
S
-4.7
-3.0
n
21
rvo
p=0.02
m
-4.8
xi
40
-5.0
-3.1
e
17
18
mo
-3.3
600
0.4
n
20
u
Cu
-5.3
n
20
plasmocytoma (MM1S)
T
-5.5
11,8
Vehicle
A
-5.8
0
8,8
8,8
0.2
Apl 100 µg/Kg x 7d/w
%
-6.5
0
6
12
48
200
Apl 140 µg/Kg x 5d/w
-7.4
NORMAL EXPRESSION
2,9
2,9
2,9
2,9
2,9
in CB17/SCID mice
-9.1
Time with Aplidin 100 nM (h)
Bortezomib 10 nM (h)
0
0.0
LOW
HIGH
- 10.4
0
5
10
15
20
25
30
10
20
30
40
50
60
- 12.1
0
- 17.6
Treatment of MM1S cells with Aplidin
Diarrhea
Nausea
Vomiting
Fatigue
Anorexia
Muscle
ALKP
CPK
AST
ALT
Days of treatment
Days of treatment
- 23.0
50 nM x 4h ( 20% apoptosis)
weakness
CONCLUSIONS
· Aplidin® is effective both as a single agent and in combination with dexamethasone in the in vitro and in vivo settings
· Its activity in relapsed/refractory MM patients is promising with an acceptable toxicity profile
Document Outline