Molecular Features of the YAP Inhibitor Verteporfin: Synthesis of Hexasubstituted Dipyrrins as Potential Inhibitors of YAP/TAZ, the Downstream Effectors of the Hippo Pathway

Article abstract of DOI:10.1002/cmdc.201700063

Porphyrin derivatives, in particular verteporfin (VP), a photosensitizer initially designed for cancer therapy, have been identified as inhibitors of the YAP–TEAD interaction and transcriptional activity. Herein we report the efficient convergent synthesis of the dipyrrin half of protoporphyrin IX dimethyl ester (PPIX-DME), in which the sensitive vinyl group was created at the final stage by a dehydroiodination reaction. Two other dipyrrin derivatives were synthesized, including dipyrrin 19 [(Z)-2-((3,5-dimethyl-4-vinyl-2H-pyrrol-2-ylidene)methyl)-3,5-dimethyl-4-vinyl-1H-pyrrole], containing two vinyl groups. We found that VP and dipyrrin 19 showed significant inhibitory effects on TEAD transcriptional activity in MDA-MB-231 human breast cancer cells, whereas other compounds did not show significant changes. In addition, we observed a marked decrease in both YAP and TAZ levels following VP treatment, whereas dipyrrin 19 treatment primarily decreased the levels of YAP and receptor kinase AXL, a downstream target of YAP. Together, our data suggest that, due to their chemical structures, porphyrin- and dipyrrin-related derivatives can directly target YAP and/or TAZ proteins and inhibit TEAD transcriptional activity.

Full text of DOI:10.1002/cmdc.201700063

Accepted Article
Title: Molecular features of the YAP inhibitor, Verteporfin - synthesis of
hexasubstituted dipyrrins as potential inhibitors of YAP/TAZ, the
downstream effectors of the Hippo pathway
Authors: FLORIANE GIBAULT, FABRICE BAILLY, matthieu corvaisier,
mathilde coevoet, guillemette huet, patricia Melnyk, and
Philippe Cotelle
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To be cited as: ChemMedChem 10.1002/cmdc.201700063
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10.1002/cmdc.201700063
ChemMedChem
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Molecular features of the YAP inhibitor, Verteporfin - synthesis of
hexasubstituted dipyrrins as potential inhibitors of YAP/TAZ, the
downstream effectors of the Hippo pathway
Floriane Gibault,[a] Fabrice Bailly,[a] Matthieu Corvaisier,[b] Mathilde Coevoet,[a] Guillemette Huet,[b]
[a]
Patricia Melnyk,[a] Philippe Cotelle
*
[a]
Ms F. Gibault, Dr. F. Bailly, Ms M. Coevoet, Prof. P. Melnyk (ORCID: 0000-0002-9555-3446), Prof. P. Cotelle (ORCID: 0000-0003-0924-0433)
Department: Onco and NeuroChemistry
Institution: University of Lille, INSERM UMR-S 1172, Jean-Pierre Aubert Research Center
Address: 3, rue du professeur Laguesse, BP 83, 59006 Lille Cedex, France
E-mail: philippe.cotelle@univ-lille1.fr
[b]
Dr. M. Corvaisier, Dr. G. Huet
Department: Mucins, epithelial differentiation and carcinogenesis
Institution: University of Lille, INSERM UMR-S 1172, Jean-Pierre Aubert Research Center
Address: Bâtiment Biserte, 1, place de Verdun, 59045 Lille Cedex, France
Abstract: Porphyrin derivatives, and in particular Verteporfin (VP), a
photosensitizer initially designed for cancer therapy, have been
identified as inhibitors of YAP-TEAD interaction and transcriptional
activity. We herein report the efficient convergent synthesis of the
dipyrrin western half-part of protoporphyrin IX dimethyl ester (PPIX-
DME) where the sensitive vinyl group was created at the final stage
by a dehydroiodation reaction. Two other dipyrrin derivatives were
synthesized including dipyrrin 19 containing two vinyl groups. We
found that VP and dipyrrin 19 showed a significant inhibitory effect on
TEAD transcriptional activity in MDA-MB-231 cells whereas other
compounds did not show significant changes. In addition, we
observed a marked decrease in both YAP and TAZ levels following
VP treatment whereas dipyrrin 19 treatment primarily reduced the
level of YAP and the receptor kinase AXL, a downstream target of
YAP. Altogether, our data suggests that in function of their chemical
structure, porphyrin- and dipyrrin-related derivatives can directly
target YAP and/or TAZ proteins and inhibits TEAD transcriptional
activity.
several drawbacks: (i) it is a photosensitizer and therefore is
highly phototoxic; (ii) it has been identified as an inhibitor of
autophagy via a low-level production of singlet oxygen [10]; (iii) it
induces the oligomerization of up to 250 proteins including 80
mitochondrial proteins [11]; (iv) it has also been reported that VP
up-regulates 14-3-3σ (a protein responsible for the cytoplasmic
retention of YAP)[12].
OH
MeOOC
MeOOC
NH HN
NH HN
NH
NH HN
NH
OH
N
N
N
N
COOR
COOH
COOR
COOR'
COOH
COOR
Verteporfin (equimolar
mixture of racemic R=H,
R'=Me and R=Me, R'=H)
Protoporphyrins IX
PPIX: R=H
PPIX-DME: R=Me
Hematoporphyrin (HP)
MeOOC
Introduction
YAP(TAZ), the downstream effectors of the Hippo pathway,
interact with TEAD to regulate stem cell proliferation, tissue
growth and organ size [4]. Oncogenic YAP/TAZ are
overexpressed in many cancers [5-8] causing overgrowth
phenotypes and metastasis. Liu-Chittenden and colleagues [9]
were the first to demonstrate to feasibility to disrupt YAP/TEAD
complex as a pertinent therapeutic strategy in cancer therapy and
identified three members of the porphyrin family, namely VP,
protoporphyrin IX (PPIX) and hematoporphyrin (HP), as inhibitors
of YAP/TEAD-dependent transcription. VP was found to be the
most effective compound to inhibit the complex between YAP and
TEAD2 and was used VP for in vivo validation. Indeed, VP
treatment efficiently suppressed liver overgrowth induced by YAP
overexpression in mice models.
Verteporfin, is a photosensitizer initially designed for cancer
therapy, which presents high cytotoxicity for tumor cells only
under light stimulation [1]. Rapidly VP was repositioned in the field
of ophthalmology and photodynamic therapy for the treatment of
age-related macular degeneration [2] (VP was FDA-approved in
2000 and marketed by Novartis under the trade name: visudyne®).
VP was recently reported to exert different biological properties
without light activation [3 and references cited therein]. However,
the use of VP as an inhibitor of YAP-induced overgrowth presents
N
N
N
N
NH
NH
NH
NH
COOMe
COOMe
Dipyrrin 1
Dipyrrin 18
Dipyrrin 19
Dipyrrin 20
Figure 1. Structures of VP, HP, PPIX and related dipyrrins
In this paper, we first focused our attention on the structural
features that confer to VP its ability to control the YAP/TEAD
interaction and decrease the induction of the main target genes.
For this purpose, we first compared the biological properties of
VP, PPIX and its dimethyl ester (Figure 1) in MDA-MB-231 breast
cancer cell line. In a second round, we synthesized the
hexasubstituted dipyrrin 1 corresponding to the western part of
PPIX dimethyl ester (PPIX-DME).
We report herein the first convergent synthesis of this dipyrrin
derivative starting from the known 2-formylpyrrole 2 (Scheme 1)
[13]. Additionally, we synthesized two other symmetric
1
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10.1002/cmdc.201700063
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hexasubstituted dipyrrins 18 and 19 and report their cellular
activities on the YAP(TAZ)/TEAD complexes. Chemistry of
dipyrrins [14] is directly connected to the synthesis of porphyrins
and the access to dipyrrinato complexes possessing remarkable
optical properties, however nothing is known about the biological
properties of this class of molecules.
The third strategy (Scheme 3) consisted in the ultimate Stille
vinylation of the iododipyrrin 10 which could be issued from the
condensation of the formylpyrrole 2 and the iodopyrrole 9 [21, 22].
Unfortunately, the condensation of the two pyrrolo units 2 and 7
yielded the unexpected deiododipyrrin 11.
The biological evaluation has been made in two distinct steps:
firstly, as a reference, we measured the effect of the inhibition of
expression YAP and/or TAZ by siRNA in MDA-MB-231 cells on
the expression of some relevant target genes (Cyr61, CTGF and
AXL). In a second time, we measured the biological effect of VP
and related protoporphyrins and dipyrrins on by using two
different methods: 1- measurement of TEAD transcriptional
activity using 8xGTIIC-luciferase reporter assay [15], 2-
evaluation of the protein levels of Cyr61, CTGF and AXL.
I
a
b
N
NH
NH
NH
H
EtOOC
HOOC
EtOOC
6
7
8
NH
c
+
OHC
1
NH
COOMe
Results and Discussion
COOMe
2
Chemistry
Scheme 2. (a) Vinyltributyltin (2 eq.), Pd(PPh3)2Cl2 (0.05 eq.), toluene, 110 °C,
2 h, 48%; (b) LiOH (8 eq.), EtOH/H2O, 90 °C, 8 h, 32%; (c) HBr 33% in AcOH
(8 eq.), MeOH, rt, 1h.
Our first strategy to obtain the dipyrrin 1 (Scheme 1) was inspired
from the total synthesis of hematoporphyrin published by Martin
et al. [16].
H
N
O
Br
NH
NH
NH
OHC
I
O
a
NH
OHC
11
COOMe
I
b
NH
O
+
COOMe
Br
H
N
NH
5'
a
+
HOOC
9
HO
NH
NH
3
COOMe
COOMe
2
2
N
N
N
N
H
COOMe
b
4
NH
NH
NH
NH
1
10
COOMe
5
COOMe
1
COOMe
COOMe
Scheme 1. (a) HBr 33% in AcOH (8 eq.), MeOH, rt, 1 h, 89%; (b) CeCl3.7H2O
Scheme 3. (a) HBr 33% in AcOH (8 eq.), MeOH, rt, 1 h, 75%; (b) Vinyltributyltin
(1 eq.), NaBH4 (2 eq.), dioxane/propan-2-ol, 100 °C, 20 h, 75%.
(2 eq.), Pd(PPh3)2Cl2 (0.05 eq.), toluene, 110 °C, 2 h.
Our fourth strategy (Scheme 4) was based on the condensation
of the formylpyrrole 2 and the iodoethylpyrrole 13 (synthesized
from the known hydroxyethylpyrrole 12 [23]). Base-induced
dehydroiodation of the yielded iodoethyldipyrrin 14 gave
successfully the target dipyrrin 1 with a moderate yield (47%).
Symmetric tetramethyldi(3-methoxy-3-oxopropyl)dipyrrin 18 and
di(2-iodoethyl)tetramethyldipyrrin 17 were also synthesized using
the same strategy than for the synthesis of 14 (Scheme 5). 18
was obtained with the same yield as recently reported by Lund
and Thompson, who synthesized symmetric dipyrrin
hydrobromides from 2-formylpyrroles [24]. The double
dehydroiodation of di(2-iodoethyl)tetramethyldipyrrin 17 using
The condensation of the two pyrrolo units 2 [17] and 3 [18] using
HBr in acetic acid gave the dipyrrin 4 with high yield (89%) but the
reduction of the acetyl group in presence of NaBH4 [15] alone or
with CeCl3 [19] yielded the dipyrromethane 5’ instead of the
expected alcohol 5.
In a second strategy (Scheme 2), we planned to obtain the target
compound 1 through the similar condensation of the pyrrolo units
2 and 8. Firstly the vinylpyrrole 8 was obtained via the Stille
vinylation [20] of the iodopyrrole 6. Unfortunately, vinylpyrrole 8
was found to be unstable under the acidic conditions required for
the dipyrrin synthesis.
2
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DBU gave tetramethyldivinyldipyrrin 19 with a modest 36% yield.
(Scheme 5).
and yielded a high expression of gene targets (AXL, Cyr61 and
CTGF).
We first analyzed the expression of mRNA (by RT-qPCR) and
proteins (Western Blot) of YAP, TAZ, AXL, Cyr61 and CTGF in
the presence of siYAP, siTAZ or both (Si Y-T). Knockdown of
either YAP or TAZ decreased the expression of gene targets
(Figure 2) but did not show any selectivity of one co-activator.
Dual YAP/TAZ knockdown significantly decreased the protein
levels of Cyr61, CTGF and AXL.
I
OH
a
HN
HN
COOtBu
COOtBu
12
13
I
+
OHC
N
N
Br
NH
H
b
c
NH
NH
COOMe
2
COOMe
COOMe
14
1
Scheme 4. (a) PPh3 (2 eq.), Imidazole (2 eq.), I2 (2 eq.), DCM, rt, 20 h, 83%; (b)
i) TFA (44 eq.), rt, 30min ii) HBr 33% in AcOH (8 eq.), MeOH, rt, 1 h, 63%; (c)
DBU (2.5 eq.), DCM, rt, 20 h, 47%.
R
I
R
OHC
N
N
N
Br
Br
NH
a
H
H
b
+
HN
NH
NH
NH
COOtBu
R
R
I
19
13 R= I
15 R=COOMe
16 R=I
2 R=COOMe
17 R=I
18 R=COOMe
17
Scheme 5. (a) HBr 33% in AcOH (8 eq.), MeOH, rt, 1 h, 65% for 17 and 68%
for 18; (b) DBU (4 eq.), DCM, rt, 20 h, 36%.
Biology
YAP and TAZ are transcriptional coactivators that function as
effectors of the Hippo pathway. To maintain Hippo pathway
homeostasis, TAZ accumulation can be negatively regulated by
YAP abundance [25]. More recently, Moroishi et al. [26] reported
a reciprocal negative regulation between these two coactivators
in MCF10A cells, through TEAD-dependent transcription.
Submitted to oxidative stress, differences were found between
YAP and TAZ. TAZ was less sensitive to oxidative stress than
YAP [27]. For these reasons, we focused our attention on the
effects of VP, PPIX and dipyrrins on the two Hippo pathway
effectors (YAP and TAZ) and on the YAP(TAZ)/TEAD gene
targets (AXL, Cyr61 and CTGF).
Figure 2. Effects of VP (10 µM), YAP, TAZ or YAP/TAZ-silencing on mRNA
expression (A) or protein production (B) of Cyr61, CTGF or AXL in MDA-MB-
231 cells. For the mRNA expression, results are the average of three
independent experiments. Data are presented as mean SEM (n = 3, *p < 0.1, **
p < 0.01, *** p < 0.001). For protein production, data are representative of 3
independent experiments.
Depending on the cell lines, the Hippo pathway effectors can be
differently expressed. This has been clearly shown on different
colon cancer cell lines by some of us [15] that HT-29 cells
expressed mainly YAP whereas HCT-116 or RKO cells
expressed both YAP and TAZ. Moreover, the relative TEAD
activity is also cell line-dependent. We selected MDA-MB-231
breast cancer cell line because it similarly expressed YAP and
TAZ co-activators, presented a high TEAD transcriptional activity
As previously reported [10], VP is a photoactivable molecule and
exposure to light must always be stringently avoided during cell
treatment. However, even in the absence of light, we observed a
decrease in YAP and TAZ levels in western blots. While the RNA
expression of YAP is not affected (Figure 2A), its protein
production decreases by 74%. This already reported
proteotoxicity [11] was also observed TAZ (Figure 2B) even if TAZ
could be considered to be less sensitive to oxidative stress
3
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conditions [27]. The reduction of YAP expression was previously
reported in hepatocellular carcinoma cells treated by a 1,2-
dithiole-3-thione derivative [28], known to generate in vitro
superoxide anion [29] and hydrogen peroxide [30].
VP (10 µM) efficiently decreased the mRNA expression of the
downstream gene targets (Figure 3A). At the protein levels, AXL,
and CTGF appeared mostly affected (Figure 3B). CTGF was
already found to be more sensitive than Cyr61 in ovarian cancer
cells treated by VP [31]. Inversely, VP affects more efficiently
Cyr61 than CTGF in retinoblastoma cells [32]. It may be
suggested that VP could also affect AXL, and CTGF levels
through its proteotoxic properties.
Using the TEAD luciferase assay, we measured the ability of VP
and related compounds to inhibit the TEAD activity (Figure 3). We
found that VP at a 10 µM concentration inhibited this activity by
50%. Tested protoporphyrins and dipyrrins did not show
significant effect except dipyrrin 19.
Conversely to the results obtained in HEK293 cells [9], we did not
observe a significant decrease of the TEAD luciferase activity
following PPIX (or PPIX-DME) treatment.
Figure 4. Effects of porphyrins (left) and dipyrrins (right) on YAP, TAZ, Cyr61,
CTGF and AXL levels. MDA-MB-231 cells were incubated in the presence of
the drug (10 µM) for 48h. Actin was used as loading control. Data are
representative of 3 independent experiments.
To test the oxidative stress/proteotoxicity hypothesis, we treated
MDA-MB-231 cells pre-incubated with 10 mM N-acetylcysteine
with VP or dipyrrin 19. No significant modifications were observed
on YAP, TAZ, AXL, CTGF and Cyr61 protein levels
(supplementary information). This result suggests that the
inhibition of YAP and TAZ by VP should not be due to ROS
production.
Conclusion
We successfully synthesized the dipyrrin western part of
protoporphyrin IX, dipyrrin 1, from the known tert-butyl 4-(2-
Figure 3. Inhibition of YAP(TAZ)/TEAD interaction by VP, protoporphyrins and
dipyrrins. MDA-MB-231 cells expressing 8xGTIIC luciferase were incubated for
48h with tested drugs (10 µM). Data are representative of three independent
experiments in duplicate (Data are presented as mean SEM (n = 6, *p < 0.1).
hydroxyethyl)-3,5-dimethyl-1H-pyrrole-2-carboxylate.
The
sensitive vinyl group was created at the final stage by a
dehydroiodation reaction. We submitted dipyrrin 1 to Diels-Alder
reaction conditions but failed to obtain the western part of VP due
to the unstability of 1 to prolonged heat at toluene boiling point.
This dipyrrin 1 could be useful to study other biological properties.
Using the same strategy, the divinyl-dipyrrin 19 was synthesized.
Independently to its mechanism of action, it has been well-
established in several animal models that VP administration
allows to control efficiently the organ growth via the Hippo
pathway. VP has been firstly reported as a YAP/TEAD complex
inhibitor whereas its ability to oligomerize several cellular proteins
could correspond to a low protein selectivity [11].
We observed that VP significantly inhibits the TEAD luciferase
activity in agreement to previously reported data [9]. However, we
also observed that VP decreases the expression of YAP and TAZ,
and such effect must result in a decrease in YAP(TAZ)/TEAD co-
transcriptional activity. It has been reported that VP can induce
the formation of high molecular weight oligomers through a cross-
We then analyzed the effect of PPIX, PPIX-DME and dipyrrin
derivatives on the expression of YAP, TAZ and their target genes.
Conversely to VP, PPIX, PPIX-DME did not induce a dramatic
decrease in YAP and TAZ (Figure 4). This may be related to the
expected weaker proteotoxicity of both compounds, which
possess a significantly lowest singlet oxygen quantum yield than
VP [3].
Dipyrrin 1, the western part of protoporphyrin IX dimethyl ester
(PPIX-DME) did not show any significant changes. But
interestingly, symmetric dipyrrin 19 containing two vinyl groups
significantly decreased the level of YAP whereas TAZ level
remained at a similar level. This result is well connected to the
fact that dipyrrin 19 is the only dipyrrin derivative able to induce a
decrease in TEAD transcriptional activity in the TEAD luciferase
assay. This derivative also resulted in a decrease in AXL level.
4
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NMR spectra were recorded on a Bruker DRX 300 spectrometer (operating
at 300 MHz for 1H and 75 MHz for 13C). Chemical shifts are expressed in
ppm relative to tetramethylsilane (TMS) or to residual proton signal in
deuterated solvents. Chemical shifts are reported as position (δ in ppm),
multiplicity (s = singulet, d = doublet, t = triplet, q = quartet, m = massif, br
= broad), coupling constant (J in Hz), relative integral and assignment. The
attributions of protons and carbons were achieved by analysis of 1D and
2D experiments (1H, 13C, COSY, HSQC and HMBC).
LC-MS were performed on a Varian triple quadrupole 1200W mass
spectrometer equipped with a non-polar C18 TSK-gel Super ODS (4.6 x
50 mm) column, using electrospray ionisation and a UV detector (diode
array). Elution was performed at a flow rate of 2 mL/min with water-formic
acid (pH = 3.8) as eluent A and ACN-formic acid (pH = 3.8) as eluent B,
employing a 0.25 min plateau with 0 % B and a linear gradient from 0 % B
to 98 % B in 3.25 min, followed by a 0.5 min plateau with 98 % B. Then,
column re-equilibration was performed for 1 min. The injection duty cycle
was 5 min, taking into account the column equilibration time.
linking of proteins potentiated by oxidative stress [10]. The
decrease in YAP and TAZ levels observed after VP treatment is
not due to oligomerization (no multimeric form of YAP or TAZ
have
been
detected)
but
might
result
from
degradation/modification of these proteins. Indeed, we always
observed on the western blots that the spot of YAP and TAZ are
always slightly higher than that observed without VP.
In addition, we identified here a compound, dipyrrin 19, able to
decrease YAP level and TEAD transcriptional activity. Such effect
of dipyrrin 19 could be due the addition of two vinyl groups to the
dipyrrin skeleton resulting in proteotoxicity.
The absence of significant modifications on YAP, TAZ, AXL,
CTGF and Cyr61 protein levels in MDA-MB-231 cells pre-
incubated with 10 mM N-acetylcysteine suggests that the
inhibition of YAP and TAZ by VP should not be due to ROS
production.
Altogether, these data suggest that in function of their chemical
structure, porphyrin- and dipyrrin-related derivatives can directly
target YAP and/or TAZ proteins and inhibits TEAD transcriptional
activity.
VP, which is mainly localized in mitochondria, might also affected
the YAP and TAZ expression via the mitochondrial fusion/Hippo
pathway cross-talking [33]. This link between Hippo pathway and
mitochondria fusion could be of significance to cancer biology
[34].
The melting point analyses were performed on Barnstead Electrothermel
Melting Point Series IA9200.
The UV-vis spectra were recorded using a NanoDrop spectrometer
(ThermoFisherScientific). Characteristic peaks are given in nm and
classified according to their relative intensity (s strong, m medium, w weak).
(Z)-4-Acetyl-2-((4-(3-methoxy-3-oxopropyl)-3,5-dimethyl-1H-pyrrol-2-
yl)methylene)-3,5-dimethyl-2H-pyrrolium bromide (4)
A solution of 33% HBr in AcOH (2.7 eq., 497 mg, 0.3 mL, 2.9 mmol) was
added dropwise to a solution of methyl 3-(5-formyl-2,4-dimethyl-1H-pyrrol-
3-yl)propanoate [17] (1 eq., 228 mg, 1.1 mmol) and 3-acetyl-2,4-
dimethylpyrrole [18] (1 eq., 150 mg, 1.1 mmol) in MeOH (3 mL) at room
temperature under nitrogen atmosphere. The mixture was stirred at room
temperature for 4 h and the product was collected by filtration. The residue
was used without further purification in the next step.
The title compound (400 mg, 0.97 mmol, 89%) was obtained as an orange
solid.
1H NMR (300 MHz), δ (ppm, CDCl3): 2.37 (s, 3 H, CH3), 2.48-2.53 (m, 5
H, CH2 + CH3), 2.59 (s, 3 H, CH3), 2.77-2.82 (m, 5 H, CH2 + CH3), 2.98 (s,
3 H, CH3), 3.68 (s, 3 H, OCH3), 7.26 (s, 1 H, CH), 13.22 (s, 1 H, NH), 13.69
(s, 1 H, NH).
Experimental Section
13C NMR (75 MHz), δ (ppm, CDCl3): 10.4 (CH3), 12.7 (CH3), 13.5 (CH3),
15.8 (CH3), 19.2 (CH2), 31.6 (CH3), 33.4 (CH2), 51.9 (OCH3), 120.6 (CH),
126.8, 127.8, 129.4, 132.7, 143.4, 144.8, 154.3, 160.5, 172.5 (CO), 203.5
(CO).
Biology
Human breast cancer cell line
MDA-MB-231 cell line was purchased from the American Type Culture
Collection and cultivated in DMEM media containing 10% of heat
inactivated FBS, L-glutamine and penicillin/streptomycin.
LC-MS (ESI) m/z Calculated: 408.10, Found: 329.16 [M-HBr+H]+; tR = 2.15
Methyl 3-(5-((4-acetyl-3,5-dimethyl-1H-pyrrol-2-yl)methyl)-2,4-dimethyl-
1H-pyrrol-3-yl)propanoate (5’)
Western blotting
Total extracts of cells were obtained with a RIPA based buffer containing
protease and phosphatase inhibitors (Roche). Western blot were carried
out using 20 μg of protein lysates with the NuPage Electrophoresis and
Iblot transfer systems (Life Technologies). β-Actin was used as loading
control for total extracts,
Dipyrrin 4 (1 eq., 0.19 g, 0.46 mmol) was dissolved in a mixture of DCM
(28 mL) and MeOH (7 mL). Sodium borohydride (5.0 eq., 88 mg, 2.3 mmol)
was added in one portion and the mixture was stirred at room temperature
for 2 h. Then the solution was washed with saturated aqueous bicarbonate
solution and brine, dried over sodium sulfate and concentrated under
vacuum. The residue was purified by flash chromatography (DCM/MeOH,
95:5 (v/v), NH3g sat).The title compound (115 mg, 0.35 mmol, 75%) was
obtained as an orange solid.
TEAD transcriptional activity
Cells at 50% confluence were transfected with the TEAD luciferase
reporter plasmid 8XGTIIC-Luciferase (Addgene reference 34615) vs a
control luciferase plasmid. After 48 hours post transfection, cells are lysed
in Reporter Lysis Buffer (Promega) and luciferase activity was measured
on the Mithras LB940 plate reader and normalized to protein concentration.
mRNA expression
mRNAs were extracted from cultured cells or human tumors with the
NucleoSpin kit (Macherey-Nagel). Retrotranscription was done on 1 μg of
mRNA accordingly to the Advantage RT-for-PCR Kit protocol (Clontech).
PCR was performed using SsoFastTM Evagreen Supermix kit following
the manufacturer’s protocol using the CFX96 real time PCR system (Bio-
Rad). To monitor any change in mRNA expression, we used the ΔΔCt
method between one condition and a control condition after normalization
with the housekeeper gene RPLP0. Each sample was done in triplicate.
1H NMR (300 MHz), δ (ppm, CDCl3): 1.98 (s, 3 H, CH3), 2.11 (s, 3 H, CH3),
2.26 (s, 3 H, CH3), 2.41 (s, 3 H, CH3), 2.43 (s, 3 H, CH3), 2.46 (t, J = 8.4
Hz, 2 H, CH2), 2.70 (t, J = 8.4 Hz, 2 H, CH2), 3.68 (s, 3 H, OCH3), 3.75 (s,
2 H, CH2), 8.05 (bs, 1 H, NH), 8.95 (bs, 1 H, NH).
13C NMR (75 MHz), δ (ppm, CDCl3): 9.5 (CH3), 11.3 (CH3), 12.3 (CH3),
15.5 (CH3), 20.6 (CH2), 22.0 (CH2), 31.2 (CH3), 35.9 (CH2), 51.9 (OCH3),
113.8, 115.7, 117.0, 121.6, 122.5, 122.8, 125.3, 135.0, 174.5 (CO), 196.5
(CO).
Ethyl 4-iodo-3,5-dimethyl-1H-pyrrole-2-carboxylate (6)
To a solution of N-chlorosuccinimide (1.2 eq., 0.96 g, 7.2 mmol) in DMF
(20 mL) was added NaI (1.2 eq., 1.08 g, 7.2 mmol) in small portions. The
resulting brown solution was stirred at room temperature for 1 h before the
slow addition at 0 °C of ethyl 3,5-dimethyl-1H-pyrrole-2-carboxylate (1 eq.,
1 g, 6.0 mmol) in DMF (20 mL). The reaction was stirred at room
temperature for 1 h. Aqueous 10% sodium thiosulfate solution (20 mL) was
added and the mixture was stirred for 1 h. The solid was collected by
filtration. The residue was used without further purification in the next step.
The title compound (1.61 g, 5.5 mmol, 92%) was obtained as a white solid.
1H NMR (300 MHz), δ (ppm, CDCl3): 1.36 (t, J = 1.7 Hz, 3 H, CH3), 2.29
(s, 3 H, CH3), 2.29 (s, 3 H, CH3), 4.33 (q, J = 1.7 Hz, 2 H, CH2), 8.96 (s, 1
H, NH).
Chemistry
Chemicals and solvents were purchased from various suppliers (Sigma-
Aldrich, Alfa Aesar, Fisher, VWR) and used without purification.
The reaction monitoring was performed by thin layer chromatography
(TLC) on Macherey-Nagel Alugram® Sil 60/UV254 (thickness 0.2 mm).
TLC were revealed by UV (λ = 254 nm) and/or the appropriate stain.
Purification of the compounds was carried out by column chromatography
(flash or manual). Manual chromatography was performed using
Macherey-Nagel silica gel (0.04-0.063 mm of particule size). Flash
chromatography was performed on a Reveleris® Flash Chromatography
System using Macherey-Nagel Chromabond flash RS columns.
13C NMR (75 MHz), δ (ppm, CDCl3): 14.3 (CH3), 14.5 (CH3), 14.7 (CH3),
60.3 (CH2), 72.0 (C), 118.3 (C), 130.7 (C), 134.6 (C), 161.4 (CO).
LC-MS (ESI) m/z Calculated: 292.99, Found: 291.99 [M-H]-; tR = 3.14 min
5
This article is protected by copyright. All rights reserved.

10.1002/cmdc.201700063
ChemMedChem
FULL PAPER
Ethyl 3,5-dimethyl-4-vinyl-1H-pyrrole-2-carboxylate (7)
triphenylphosphine (2 eq., 219 mg, 0.84 mmol) were dissolved in DCM (5
mL) under a nitrogen atmosphere at room temperature. After complete
dissolution, iodine (2 eq., 212 mg, 0.84 mmol) was added portionwise at 0
°C. The mixture was stirred overnight at room temperature. The reaction
was quenched with aqueous 10% Na2S2O3 solution (10 mL). The aqueous
layer was extracted with DCM (3 x 10 mL), dried over Na2SO4 and
concentrated under vacuum. The residue was purified by flash
chromatography (Cyclohexane/EtOAc, 7:3 (v/v)).
A mixture of ethyl 4-iodo-3,5-dimethyl-1H-pyrrole-2-carboxylate (1 eq.,
1.00 g, 3.4 mmol) , vinyltributyltin (2 eq., 2.16 g, 2.0 mL, 6.8 mmol) and
bis(triphenylphosphine)palladium(II) dichloride (5 %, 0.12 g, 0.17 mmol) in
toluene (50 mL) was refluxed for 2 h under a nitrogen atmosphere. During
the course of the reaction, the color changed from yellow to black as Pd°
was formed. The reaction was cooled to room temperature and DCM (50
mL) was added. The resulting solution was washed with satured aqueous
NaCl (50 mL) and water (50 mL). The organic phase was separated, dried
over Na2SO4 and concentrated under vacuum. The residue was purified
by flash chromatography (DCM/MeOH, 95:5 (v/v)).
The title compound (138 mg, 0.4 mmol, 95%) was obtained as a white
solid.
1H NMR (300 MHz), δ (ppm, CDCl3): 1.56 (s, 9 H, 3 CH3), 2.22 (s, 3 H,
CH3), 2.24 (s, 3 H, CH3), 2.95 (t, J = 7.9 Hz, 2 H, CH2), 3.14 (t, J = 8.3 Hz,
2 H, CH2), 8.64 (s, 1 H, NH).
The title compound (317 mg, 1.6 mmol, 48%) was obtained as a dark
brown solid.
1H NMR (300 MHz), δ (ppm, CDCl3): 1.36 (t, J = 7.1 Hz, 3 H, CH3), 2.32
(s, 3 H, CH3), 2.39 (s, 3 H, CH3), 4.31 (q, J = 7.1 Hz, 2 H, CH2), 5.17 (dd,
J = 1.7 Hz, Jcis = 11.6 Hz, 1 H, CH2), 5.28 (dd, J = 1.7 Hz, Jtrans = 17.9 Hz,
1 H, CH2), 6.59 (dd, Jtrans = 17.9 Hz, Jcis = 11.6 Hz, 1 H, CH), 8.62 (s, 1 H,
NH).
13C NMR (75 MHz), δ (ppm, CDCl3): 5.6 (CH2), 10.6 (CH3), 11.6 (CH3),
28.5 (3 CH3), 29.3 (CH2), 80.4 (C), 118.5 (C), 121.0 (C), 125.7 (C), 129.1
(C), 161.1 (CO).
LC-MS (ESI) m/z Calculated: 349.05, Found: 295.07 [M-tBu+H]+, tR = 3.39
min
13C NMR (75 MHz), δ (ppm, CDCl3): 11.2 (CH3), 12.8 (CH3), 14.5 (CH3),
59.8 (CH2), 112.7 (CH2), 125.3 (CH), 128.2 (C), 129.0 (C), 133.6 (C), 139.3
(C), 161.6 (CO).
(Z)-4-(2-Iodoethyl)-2-((4-(3-methoxy-3-oxopropyl)-3,5-dimethyl-1H-pyrrol-
2-yl)methylene)-3,5-dimethyl-2H-pyrrolium bromide (14)
TFA (44.4 eq., 1.15 mL, 25.5 mmol) was added dropwise to a solution of
tert-butyl 4-(2-iodoethyl)-3,5-dimethyl-1H-pyrrole-2-carboxylate (1 eq.,
200 mg, 0.57 mmol) at room temperature under a nitrogen atmosphere.
The solution was stirred for 30 min. Then, a solution of methyl 3-(5-formyl-
2,4-dimethyl-1H-pyrrol-3-yl)propanoate (1 eq., 119 mg, 0.57 mmol) in
MeOH (2.3 mL) was added dropwise, followed by 33% HBr solution in
AcOH (8.99 eq., 1263 mg, 0.86 mL, 5.1 mmol). The mixture was stirred at
room temperature for 1 h and the product was collected by filtration. The
residue was used without further purification in the next step.
The title compound (200 mg, 0.38 mmol, 67%) was obtained as an orange
solid.
3,5-Dimethyl-4-vinyl-1H-pyrrole-2-carboxylic acid (8)
Ethyl 4-ethenyl-3,5-dimethyl-1H-pyrrole-2-carboxylate (1 eq., 158 mg,
0.82 mmol) and LiOH (8 eq., 156 mg, 6.5 mmol) were dissolved in EtOH
(2 mL) and H2O (2 mL) . The solution was heated at 90 °C for 1 h. The
solvent was evaporated under vacuum. The residue was taken up in
EtOAc (5 mL) and water (5 mL). The aqueous layer was acidified until pH
5.0 with H3BO3 (pH 5.1) or a few drops of concentrate H2SO4 before being
extracted with EtOAc. Organics layers were dried over Na2SO4 and
concentrated under vacuum. The residue was used without further
purification in the next step.
The title compound (43 mg, 0.3 mmol, 32%) was obtained as a dark brown
1H NMR (300 MHz), δ (ppm, CDCl3): 2.31 (s, 6 H, 2 CH3), 2.47 (t, J = 7.4
Hz, 2 H, CH2), 2.71 (s, 6 H, 2 CH3), 2.76 (t, J = 7.3 Hz, 2 H, CH2), 3.02 (t,
J = 6.9 Hz, 2 H, CH2), 3.19 (t, J = 6.9 Hz, 2 H, CH2), 3.67 (s, 3 H, CH3),
7.08 (s, 1 H, CH), 13.11 (s, 1 H, NH).
solid.
1H NMR (300 MHz), δ (ppm, Acetone-d6): 2.32 (s, 3 H, CH3), 2.36 (s, 3 H,
CH3), 5.07 (dd, J = 1.8 Hz, Jcis = 11.6 Hz, 1 H, CH2), 5.25 (dd, J = 1.8 Hz,
Jtrans = 17.9 Hz, 1 H, CH2), 6.61 (dd, Jtrans = 17.9 Hz, Jcis = 11.6 Hz, 1 H,
CH), 10.24 (s, 1 H, NH).
13C NMR (75 MHz), δ (ppm, CDCl3): 3.4 (CH2), 10.2 (CH3), 10.4 (CH3),
13.0 (2 CH3), 19.3 (CH2), 28.3 (CH2), 33.7 (CH2), 51.8 (OCH3), 119.4 (CH),
126.0 (C), 126.4 (C), 127.3 (C), 127.4 (C), 142.1 (C), 142.9 (C), 153.4 (C),
155.0 (C), 172.6 (CO).
13C NMR (75 MHz), δ (ppm, CDCl3): 10.5 (CH3), 11.5 (CH3), 111.0 (CH2),
117.7 (C), 126.0 (C) 129.4 (CH), 131.2 (C), 132.6 (C), 164.1 (CO).
LC-MS (ESI) m/z Calculated: 520.02, Found: 441.03 [M-HBr+H]+, tR = 2.73
min
4-iodo-3,5-dimethyl-1H-pyrrole-2-carboxylic acid (9)
Ethyl 4-iodo-3,5-dimethyl-1H-pyrrole-2-carboxylate (1 eq., 500 mg, 1.7
mmol) and LiOH (8 eq., 326 mg, 13.6 mmol) were dissolved in EtOH (5
mL) and H2O (5 mL). The solution was heated at 90 °C for 1 h. The solvent
was evaporated under vacuum. The residue was taken up in EtOAc (50
mL) and water (50 mL). The aqueous layer was acidified to pH 5.0 with
H3BO3 (pH 5.1) or few drops of concentrate H2SO4 before being extracted
with EtOAc. Organics layers were dried over sodium sulfate and
concentrated under vacuum. The residue was purified by flash
chromatography (DCM/MeOH, 95:5 (v/v)).
(Z)-Methyl
dimethyl-1H-pyrrol-3-yl)propanoate (1)
3-(5-((3,5-dimethyl-4-vinyl-2H-pyrrol-2-ylidene)methyl)-2,4-
(Z)-4-(2-Iodoethyl)-2-{[4-(3-methoxy-3-oxopropyl)-3,5-dimethyl-1H-pyrrol-
2-yl]methylidene}-3,5-dimethyl-2H-pyrrol-1-ium bromide (1 eq., 510 mg,
0.98 mmol) was dissolved in dry DCM (10.9 mL) under nitrogen
atmosphere. DBU (2.5 eq., 0.36 mL, 2.4 mmol) was added dropwise at
room temperature and the mixture was heated at 50 °C. After complete
reaction, the solvent was evaporated under vacuum and the residue was
purified by flash column chromatography with DCM/MeOH (95:5).
The title compound (143 mg, 0.45 mmol, 47%) was obtained as a dark
brown solid.
The title compound (399 mg, 1.5 mmol, 88%) was obtained as a dark pink
solid.
1H NMR (300 MHz), δ (ppm, Acetone-d6): 2.25 (s, 3 H, CH3), 2.27 (s, 3 H,
CH3), 10.82 (s, 1 H, OH).
UV-vis (Acetonitrile) λmax (nm) 490 (s), 370 (w), 291 (w), 224 (m).
1H NMR (300 MHz), δ (ppm, CDCl3): 2.32 (s, 3 H, CH3), 2.37 (s, 3 H, CH3),
2.47 (t, J = 1.5 Hz, 2 H, CH2), 2.72 (s, 3 H, CH3), 2.76 (t, J = 1.5 Hz, 2 H,
CH2), 2.80 (s, 3 H, CH3), 5.40 (dd, J = 1.1 Hz, Jcis = 7.7 Hz, 1 H, CH2),
5.45 (dd, J = 1.1 Hz, Jtrans = 14.1 Hz, 1 H, CH2), 6.54 (dd, Jcis = 11.6 Hz,
Jtrans = 17.9 Hz, 1 H, CH), 7.12 (s, 1 H, CH), 12.92 (s, 1 H, NH).
13C NMR (75 MHz), δ (ppm, CDCl3): 10.3 (CH3), 11.0 (CH3), 13.0 (CH3),
14.0 (CH3), 19.3 (CH2), 33.7 (CH2), 51.8 (OCH3), 117.4 (CH2), 119.4 (CH),
125.8 (C), 126.1 (C), 126.5 (C), 126.7 (C), 127.4 (CH), 141.0 (C), 142.9
(C), 154.0 (C), 155.1, (C) 172.6 (CO).
13C NMR (75 MHz), δ (ppm, CDCl3): 13.1 (CH3), 13.7 (CH3), 70.8 (C),
129.7 (C), 134.3 (C), 134.3 (C), 161.0 (CO).
LC-MS (ESI) m/z Calculated: 264.96, Found: 263.92 [M-H]-; tR = 2.50 min
(Z)-3-((4-(3-methoxy-3-oxopropyl)-3,5-dimethyl-1H-pyrrol-2-
yl)methylene)-2,4-dimethyl-3H-pyrrol-1-ium bromide (11)
A solution of 48% HBr in water (2.7 eq., 0.18 mL, 1.6 mmol) was added
dropwise to a solution of pyrrole 2 (1 eq., 0.12 g, 0.6 mmol) and pyrrole 9
(1 eq., 0.16 g, 0.6 mmol) in methanol (1.5 mL).The mixture was stirred for
1 h at room temperature and concentrated under vacuum. The residue
was purified by flash chromatography (DCM/MeOH, 95:5 (v/v)).
The title compound (65 mg, 0.2 mmol, 30%) was obtained as a dark brown
solid.
LC-MS (ESI) m/z Calculated: 312.18, Found: 313.14 [M+H]+, tR =2.65 min
4-(2-Iodoethyl)-3,5-dimethyl-1H-pyrrole-2-carbaldehyde (16)
TFA (9 eq., 0.59 mL, 7.7 mmol) was added dropwise to a solution of tert-
butyl 4-(2-iodoethyl)-3,5-dimethyl-1H-pyrrole-2-carboxylate (1 eq., 0.3 g,
0.9 mmol) in DCM (4 mL) at 0 °C under a nitrogen atmosphere. Trimethyl
orthoformate (5 eq., 0.47 mL, 4.3 mmol) was added dropwise and the
reaction was stirred under N2 for 20 min at 0 °C. The solution was warmed
to room temperature and stirred for 30 min. The mixture was neutralized
carefully with aqueous 10% NaHCO3 solution and extracted with DCM (3
1H NMR (300 MHz), δ (ppm, CDCl3): 2.25 (s, 3 H, CH3), 2.29 (s, 3 H, CH3),
2.38 (t, J = 7.3 Hz, 2 H, CH2), 2.58 (bs, 6 H, 2 CH3), 2.68 (t, J = 7.3 Hz, 2
H, CH2), 3.60 (s, 3 H, OCH3), 6.08 (s, 1 H, CH), 7.01 (s, 1 H, CH), 13.00
(bs, 1 H, NH).
13C NMR (75 MHz), δ (ppm, CDCl3): 10.2 (CH3), 12.1 (CH3), 12.8 (CH3),
14.4 (CH3), 14.2 (CH2), 33.7 (CH2), 51.8 (OCH3), 117.2 (CH), 119.6 (CH),
126.2 (C), 126.7 (C), 127.0 (C), 142.7 (C), 145.6 (C), 154.5 (C), 154.7 (C),
172.6 (CO).
x
20 mL). The combined organic layers dried over Na2SO4 and
concentrated under vacuum. The residue was purified by flash column
chromatography with Cyclohexane/EtOAc (8:2).
The title compound (150 mg, 0.5 mmol, 63 %) was obtained as light brown
solid.
Tert-butyl 4-(2-iodoethyl)-3,5-dimethyl-1H-pyrrole-2-carboxylate (13)
Tert-butyl 4-(2-hydroxyethyl)-3,5-dimethyl-1H-pyrrole-2-carboxylate (1
eq., 100 mg, 0.42 mmol) , imidazole (2 eq., 56.9 mg, 0.84 mmol) and
6
This article is protected by copyright. All rights reserved.

10.1002/cmdc.201700063
ChemMedChem
FULL PAPER
1H NMR (300 MHz), δ (ppm, CDCl3): 2.29 (s, 3 H, 3 CH3), 2.30 (s, 3 H,
CH3), 2.97 (t, J = 7.9 Hz, 2 H, CH2), 3.17 (t, J = 7.4 Hz, 2 H, CH2), 9.47
(s, 1 H, CHO), 10.25 (s, 1 H, NH).
SIRIC OncoLille and le Cancéropôle Nord-Ouest. The 300 MHz
NMR facilities were funded by the Région Hauts de France
(France), the Ministère de la Jeunesse, de l’Education Nationale
et de la Recherche (MJENR) and the Fonds Européens de
Développement Régional (FEDER).
13C NMR (75 MHz), δ (ppm, CDCl3): 4.9 (CH2), 9.07 (CH3), 11.8 (CH3),
28.7 (CH2), 117.6 (Cquat), 122.0 (Cquat), 128.0 (Cquat), 139.5 (Cquat),
176.3 (CHO).
(Z)-4-(2-Iodoethyl)-2-((4-(2-iodoethyl)-3,5-dimethyl-1H-pyrrol-2-
yl)methylene)-3,5-dimethyl-2H-pyrrolium bromide (17)
Keywords: Dipyrrins • verteporfin • non-photoinduced therapy •
Hippo pathway • cancer
TFA (27 eq., 1.1 mL, 14.5 mmol) was added dropwise to a solution of tert-
butyl 4-(2-iodoethyl)-3,5-dimethyl-1H-pyrrole-2-carboxylate (1 eq., 0.19 g,
0.5 mmol) at room temperature under a nitrogen atmosphere. The solution
was stirred for 30 min. Then a solution of 4-(2-iodoethyl)-3,5-dimethyl-1H-
pyrrole-2-carbaldehyde (1 eq., 0.15 g, 0.5 mmol) in MeOH (2 mL) was
added dropwise, followed by 33% HBr in AcOH (8.6 eq., 0.81 mL, 4.6
mmol). The mixture was stirred at room temperature for 1 h and the product
was collected by filtration. The residue was used without further purification
in the next step.
References
[1]
A.M. Richter, E. Waterfield, A.K. Jain, B. Allison, E.D. Sternberg, D.
Dolphin, J.G. Levy, Br. J. Cancer 1991, 63, 87-93.
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Med. Chem., 2016, 23, 1171-1184.
The title compound (0.2 g, 0.3 mmol, 65%) was obtained as a dark red
solid.
1H NMR (300 MHz), δ (ppm, CDCl3): 2.31 (s, 3 H, 3 CH3), 2.70 (s, 3 H,
CH3), 3.02 (t, J = 7.1 Hz, 2 H, CH2), 3.19 (t, J = 7.4 Hz, 2 H, CH2), 7.10 (s,
1 H, CH), 13.12 (s, 1 H, NH).
[4] S. Piccolo, S. Dupont, M. Cordenonsi, Physiol. Rev. 2014, 94, 1287-1312.
[5] K.F. Harvey, X. Zhang, D.M. Thomas, D.M. Nat. Rev. Cancer 2013, 13,
246-257.
13C NMR (75 MHz), δ (ppm, CDCl3): 3.3 (2 CH2), 10.5 (2 CH3), 13.1 (2
CH3), 28.3 (2 CH2), 119.7 (CH), 126.3 (2 C), 127.7 (2 C), 142.6 (2 C), 154.3
(2 C).
[6] T. Moroishi, C.G. Hansen, K.L. Guan, Nat. Rev. Cancer 2015, 15, 73-79
[7] Y. Ma, Y. Yang, F. Wang, Q. Wei, H. Qin, Int. J. Cancer 2015, 137, 2275-
86.
(Z)-4-(3-Methoxy-3-oxopropyl)-2-((4-(3-methoxy-3-oxopropyl)-3,5-
dimethyl-1H-pyrrol-2-yl)methylene)-3,5-dimethyl-2H-pyrrolium
(18)
bromide
[8] D. Kong, Y. Zhao, T.; Men, C.B. Teng, J. Drug Target 2015, 23, 125-133.
[9] Y. Liu-Chittenden, B. Huang, J.S. Shim, Q. Chen, S.J. Lee, R.A. Anders,
J.O. Liu, D. Pan, Genes Dev. 2012, 26, 1300-1305.
TFA (44 eq., 1.56 mL, 21.1 mmol) was added dropwise to a solution of 4-
(3-methoxy-3-oxopropyl)-3,5-dimethyl-1H-pyrrole-2-carboxylic acid (1 eq.,
110 mg, 0.50 mmol) at room temperature under a nitrogen atmosphere.
The solution was stirred for 30 min. Then, a solution of methyl 3-(5-formyl-
2,4-dimethyl-1H-pyrrol-3-yl)propanoate (1 eq., 100 mg, 0.50 mmol) in
MeOH (2.0 mL) was added dropwise, followed by 33% HBr solution in
AcOH (8.6 eq., 1.13 g, 0.81 mL, 4.6 mmol). The mixture was stirred at
room temperature for 1 h and the product was collected by filtration. The
residue was used without further purification in the next step.
The title compound (192 mg, 0.40 mmol, 63%) was obtained as a brown
solid.
[10] E. Donohue, A.D. Balgi, M. Komatsu, M. Roberge, PLoS One 2014, 9,
e114964.
[11] H. Zhang, S.K. Ramakrishnan, D. Triner, B. Centofanti, D. Maitra, B.
Gyorffy, J.S. Sebolt-Leopold, M.K. Dame, J. Varani, D.E. Brenner, E.R.
Fearon, M.B. Omary, Y.M. Shah, Science Signaling, 2015, 8, RA98
[12] C. Wang, X. Zhu, W. Feng, Y. Yu, K. Jeong, W. Guo, Y. Lu, G.B. Mills,
Amer. J. Cancer Res. 2016, 6, 27-37.
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[14] D.F. Taber, K. Nakajima, J. Org. Chem. 2001, 66, 2515-2517.
[15] M. Corvaisier, M. Bauzone, F. Corfiotti, F. Renaud, M. El Amrani, D. Monté,
S. Truant, E. Leteurtre, P. Formstecher, I. Van Seuningen, C. Gespach,
G. Huet, Oncotarget 2016, 7, 56700-56712.
UV-vis (Acetonitrile) λmax (nm) 480 (s), 392 (w), 226 (m).
1H NMR (300 MHz), δ (ppm, CDCl3): 2.29 (s, 6 H, 2 CH3), 2.47 (t, J = 7.5
Hz, 2 H, CH2), 2.69 (s, 6 H, 2 CH3), 2.76 (t, J = 7.5 Hz, 2 H, CH2), 3.69 (s,
6 H, 2 OCH3), 7.05 (s, 1 H, CH), 13.05 (bs, 1 H, NH).
13C NMR (75 MHz), δ (ppm, CDCl3): 10.2 (2 CH3), 12.9 (2 CH3), 19.3 (2
CH2), 33.8 (2 CH2), 51.8 (2 OCH3), 119.2 (CH), 126.1 (2 C), 126.9 (2 C),
143.2 (2 C), 154.2 (2 C), 172.7 (2 CO).
[16] P. Martin, M. Mueller, D. Flubacher, A. Boudier, H.U. Blaser, D. Spielvogel,
Organic Process Research & Development, 2010, 14, 799-804.
[17] D.A. May, T.D. Lash, J. Org. Chem., 1992, 57, 4820-4828.
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[19] O. Jacquet, T. Bergholz, C. Magnier-Bouvier, M. Mellah, R. Guillot, J.C.
Fiaud, Tetrahedron, 2010, 66, 222-226.
LC-MS (ESI) m/z Calculated: 452.13, Found: 373.19 [M-HBr+H]+, tR = 2.52
min.
(Z)-2-((3,5-Dimethyl-4-vinyl-2H-pyrrol-2-ylidene)methyl)-3,5-dimethyl-4-
vinyl-1H-pyrrole (19)
[20] J. Xang, I. Scott, Tetrahedon Lett., 1995, 36, 7043-7046.
[21] A. Treibs, H.G. Kolm, Justus Liebigs Annalen der Chemie.
1958, 614, 176-198.
(Z)-4-(2-Iodoethyl)-2-{[4-(2-iodoethyl)-3,5-dimethyl-1H-pyrrol-2-
yl]methylidene}-3,5-dimethyl-2H-pyrrol-1-ium bromide (1 eq., 170 mg, 0.3
mmol) was dissolved in dry DCM (3 mL) under a nitrogen atmosphere.
DBU (4 eq., 0.172 mL, 1.2 mmol) was added dropwise to the solution at
room temperature and the mixture was heated at 50 °C. After complete
reaction, the solvent was evaporated under vacuum and the solid residue
was purified by flash column chromatography with DCM/MeOH (95:5).
The title compound (26 mg, 0.1 mmol, 36%) was obtained as a brown
solid.
[22] S.E. Boiadjiev, D.A. Lightner, J. Org. Chem., 2003, 68, 7591-7604.
[23] P.J. Clewlow, A.H. Jackson, J. Chem. Soc., Perkin Transactions 1,
1990, 1925-1936.
[24] K.A.R. Lund, A. Thompson, Synlett. 2014, 25, 1142-1144.
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Yeoh, B.A. Callus, J. Biol. Chem. 2015, 290, 27928-38.
[26] T. Moroishi, H.W. Park, B. Qin, Q. Chen, Z. Meng, S.W. Plouffe, K.
Taniguchi, F.X. Yu, M. Karin, D. Pan, K.L. Guan, Genes Dev. 2015, 29,
1271-84.
UV-vis (Acetonitrile) λmax (nm) 506 (s), 366 (s), 292 (s), 230 (s).
1H NMR (300 MHz), δ (ppm, CDCl3): 2.40 (s, 6 H, 2 CH3), 2.83 (s, 6 H, 2
CH3), 5.36-5.49 (m, 4 H, 2 CH2), 6.53 (dd, Jcis = 11.9 Hz, Jtrans = 17.6 Hz,
2 H, 2 CH) 7.19 (s, 1 H, CH), 12.66 (s, 1 H, NH).
13C NMR (75 MHz), δ (ppm, CDCl3): 11.1 (2 CH3), 14.2 (2 CH3), 117.8 (2
CH2), 119.7 (CH), 126.5 (2 CH), 137.5 (2 C), 141.8 (2 C), 154.0 (2 C),
154.9 (2 C).
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Table of Contents
We herein report the efficient convergent synthesis of the hexasubstituted dipyrrins structurally related to verteporfin where the sensitive vinyl
group was created at the final stage by a dehydroiodation reaction. VP showed a direct effect on YAP and TAZ levels whereas dipyrrin 19
showed a direct effect only on YAP level. Altogether, these data suggests that dipyrrin-related compounds may directly affect the expression of
YAP and/or TAZ.
9
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