2. Small molecule biopharmaceutical company
Established in 2003
Located in Malvern, PA
Current therapies focus on Oncology and Infectious
Disease
Two clinical stage product candidates: Birinapant and
SHAPE
3. Birinapant
SMAC (Second
Mitochondrial Activator of
Caspases) mimetic
Triggers apoptosis through
binding to cIAP1 (Cellular
Inhibitor of Apoptosis
Protein 1) in the TNF
(tumor necrosis factor)
receptor complex
Ongoing Clinical Trials in
Hepatitis B
SHAPE
Suberohydroxamic acid (4-
methoxycarbonyl) phenyl
ester
HDAC (Histone
Deacetylase) inhibitor
Analog of SAHA
(Vorinostat)
Topical product- avoids
systemic toxicity
Ongoing Clinical Trials in
CTCL
http://tetralogicpharma.com/science/
4. Birinapant is an analog of SMAC
Condon, S., et al. J Med Chem (2014)
SMAC interacts with the BIR3
region of cIAP1 through N-
terminal AVPI sequence
Binding of SMAC to cIAP1
induces dimerization and
activation of cIAP1
Activation of cIAP1 causes
autocatalytic destruction of cIAP1
and triggers apoptosis
Benetatos, C., et al. Mol. Cancer Ther. (2014)
NH
NH
F
F
N
N
O
O
N
H
H
N
H3C
H
N
N
H
OH
OH
O
O
H3C
BIR3BIR3
Birinapant
5. Generate novel analogs of Birinapant, with
modifications to both the biindole core and peptide
sidechain
Examine the feasibility of a new dimerization
strategy towards Birinapant
6. Birinapant
O
O
O
O
Cbz = Boc =
CA B
NH
F
NH
F
CbzN
CbzN
OAc
OAc
NH
F
NH
F
N
N
OAc
O
O
N
H
Boc
N
H3C
H
N
N
Boc
H3C
O
O
NH
F
NH
F
N
N
OAc
OAc
O
O
N
H
Boc
H
N
Boc
OAc
7. NH
NH
CbzN
CbzN
OAc
OAc
R
R
1. H2, Pd/C,
THF/MeOH
HO
O
R1
H
N
Cbz
HATU, DIPEA
NMP
2.
NH
NH
N
N
OAc
OAc
R
R
O
R1
N
H
O
R1
H
N
Cbz
Cbz
NH
NH
N
N
OAc
OAc
R
R
O
R1
N
H
O
R1
H
N
O
O
Boc
N
H3C
N
Boc
H3C
1. H2, Pd/C,
THF/MeOH
HO
O
Boc
N
HATU, DIPEA
NMP
2.
NH
NH
N
N
OAc
OAc
R
R
O
R1
N
H
O
R1
H
N
O
O
Boc
N
H3C
N
Boc
H3C
1. TFA, DCM
2. aq. NaOH, MeOH
NH
NH
N
N
OH
OH
R
R
O
R1
N
H
O
R1
H
N
O
O
H
N
H3C
N
H
H3C
1 2 3
54
8. N H
N H
C b z N
C b z N
O A c
O A c
N H
N H
N
N
O H
O
O
R 1
N
H
H
N
H 3 C
H
N
N
H
H 3 C
O
O R 2
N H
N H
C b z N
B r H H N
O A c
O A c
3 3 % H B r / H O A c , D C M
R R R
R R R
O H
3 1 %
6 7 8
9. Competitive Inhibition Assay
Measures fluorescence of ARPFK-5-FAM peptide once
displaced from BIR3 region
*Mean result of four assays
Entry cIAP1, IC50 (nM)* Notes
Analog A 40 Symmetric
Analog B 54.5 Symmetric
Analog C 23.5 Asymmetric
Birinapant 50.0 -
Negative Control >1000 -
Positive Control 16.5 -
10. N H
F
N H
F
C b z N
O A c
N H
F
C b z N
O H
N H
F
O
C b z N
C l
O T B S
O
C b z N
C b z N
O A c
O A c
N H
F
N H
F
N
N
O A c
O
O
N
H
B o c
N
H 3 C
H
N
N
B o c
H 3 C
O
O
N H
F
N H
F
N
N
O A c
O A c
O
O
N
H
B o c
H
N
B o c
C b z N
O H
O H
O
O A c
C
Birinapant
A B
D E F G
O
O
Si
O
O
Cbz = Boc =TBS =
11. N H
F
N H
F
C b z N
O A c
N H
F
C b z N
O H
N H
F
O
C b z N
C l
O T B S
O
C b z N
C b z N
O A c
O A c
N H
F
N H
F
N
N
O A c
O
O
N
H
B o c
N
H 3 C
H
N
N
B o c
H 3 C
O
O
N H
F
N H
F
N
N
O A c
O A c
O
O
N
H
B o c
H
N
B o c
C b z N
O H
O H
O
O A c
Birinapant
D E F G
CA B
12. D
H I
A
N H
F
N H
F
N
N
O
O
N
H
C b z
N
H 3 C
H
N
N
C b z
H 3 C
O
O
C b zN
N H
F
N
N H
F
O
N
H
C b z
R
R
R
R
N
N H
F
O
N
H
R
C b z
N
O
H 3 C
14. 11 12
13
NH
NH
F
F
1. LiBH4, MSA,
THF TFACbzN
NH
F
O
CbzN
NH
F CbzN
CbzN
95%
61%, 2 Steps
NH
NH
F
F
CbzN
CbzN
H
H
DDQ, MeOH
14
15. 14 15 16
H O
O H
N
C b z
H A T U , D IP E A
N M P
1 . H 2 , P d /C ,
T H F /M e O H
2 .
N H
N H
F
F
C b z N
C b z N
N H
N H
F
F
N
N
O
O
N
H
H
N
C b z
C b z
H O
O C b z
N
C H 3
H A T U , D IP E A
N M P
1 . H 2 , P d /C ,
T H F /M e O H
2 .
N H
N H
F
F
N
N
O
O
N
H
H
N
C b z
N
H 3 C
N
C b z
H 3 C
O
O
16. CbzN
NH
F
1. H2, Pd/C,
THF/MeOH
HO
O
H
N
Cbz
HATU, DIPEA
NMP
2.
1. TFA
2. DDQ, MeOH
NH
NH
F
F
N
N
O
O
N
H
H
N
Cbz
Cbz
N
NH
F
O
N
H
Cbz
52%
Successful synthesis of 15
based on 1H-NMR, 13C-
NMR, LC-MS
12 17
15
17. 1. H2, Pd/C,
THF/MeOH
HO
O Cbz
N
CH3
HATU, DIPEA
NMP
2.
NH
F
NH
F
N
N
O
O
N
H
Cbz
N
H3C
H
N
N
Cbz
H3C
O
O
1. TFA
2. DDQ, MeOHN
NH
F
O
N
H
Cbz N
NH
F
O
N
H
O
Cbz
N
H3C
14%
Mix of biindole products formed
at DDQ oxidation step
16
1817
18. C:Xcalibur...060315KEs177-009ali1 6/3/2015 11:58:20 AM
RT: 0.00 - 7.49
0 1 2 3 4 5 6 7
Time (min)
0
50000
100000
150000
200000
250000
300000
350000
400000
450000
500000
550000
600000
uAU
RT: 3.53
AA: 1718322
RT: 6.59
AA: 1553924
NL:
6.14E5
Channel A
UV
Genesis
KEs177-
009ali1
C:Xcalibur...061715KES177-021ali2 6/17/2015 8:54:03 AM
RT: 0.00 - 7.49
0 1 2 3 4 5 6 7
Time (min)
0
200000
400000
600000
800000
1000000
1200000
1400000
1600000
1800000
uAU
RT: 3.49
AA: 9619141
RT: 6.06
AA: 9770391
NL:
1.96E6
Channel A
UV
Genesis
KES177-
021ali2
C:Xcalibur...060515KES177-012ali2 6/5/2015 12:21:41 PM
RT: 0.00 - 7.49
0 1 2 3 4 5 6 7
Time (min)
-40000
-20000
0
20000
40000
60000
80000
100000
120000
140000
160000
180000
200000
220000
240000
uAU
RT: 3.52
AA: 672813
RT: 5.84
AA: 808489
NL:
2.45E5
Channel A
UV
Genesis
KES177-
012ali2
RT: 6.59
RT: 5.84
RT: 5.80
RT: 5.44
RT: 6.02
14
15
16
N H
F
N H
F
N
N
O
O
N
H
C b z
N
H 3 C
H
N
N
C b z
H 3 C
O
O
N H
N H
F
F
N
N
O
O
N
H
H
N
C b z
C b z
N H
N H
F
F
C b z N
C b z N
19. Purification techniques applied:
Reverse and Normal Phase HPLC (High Pressure Liquid
Chromatography)
Silica gel flash chromatography / TLC (Thin Layer
Chromatography)
Recrystallization
Analysis methods:
LC-MS (Liquid Chromatography-Mass Spectrometry)
1H-NMR
13C-NMR
COSY (Correlation Spectroscopy) NMR
Participant in CRO teleconference meetings
Kept a laboratory notebook in following GLP
Learned basics of SMAC mimetic use in therapeutics
20. Thomas Haimowitz, PhD
Stephen Condon, PhD
Christina DiMarco
Yijun Deng, PhD
Tanya Vakhilt
TetraLogic Team
West Chester University PPD Program
