A Novel Class of N-Sulfonyl and N-Sulfamoyl Noscapine Derivatives that Promote Mitotic Arrest in Cancer Cells

Article abstract of DOI:10.1002/cmdc.201900477

Noscapine displays weak anticancer efficacy and numerous research efforts have attempted to generate more potent noscapine analogues. These modifications included the replacement of the N-methyl group in the 6′-position with a range of substituents, where N-ethylcarbamoyl substitution was observed to possess enhanced anticancer activity. Herein, we describe advances in this area, namely the synthesis and pharmacological evaluation of a series of N-sulfonyl and N-sulfamoyl noscapine derivatives. A number of these sulfonyl-containing noscapinoids demonstrated improved activities compared to noscapine. ((R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-methoxy-6-((1-methyl-1H-imidazol-4-yl)sulfonyl)-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]isoquinoline) (14 q) displayed sub-micromolar activities of 560, 980, 271 and 443 nM against MCF-7, PANC-1, MDA-MB-435 and SK-MEL-5 cells, respectively. This antiproliferative effect was also maintained against drug-resistant NCI/Adr^RES cells despite high expression of the multidrug efflux pump, P-glycoprotein.

Full text of DOI:10.1002/cmdc.201900477

DOI: 10.1002/cmdc.201900477
Full Papers
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A Novel Class of N-Sulfonyl and N-Sulfamoyl Noscapine
Derivatives that Promote Mitotic Arrest in Cancer Cells
[a]
[a]
[b]
[b]
[b]
Cassandra Yong, Shane M. Devine, Xuexin Gao, Angelina Yan, Richard Callaghan,
[a]
[a]
Ben Capuano, and Peter J. Scammells*
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Noscapine displays weak anticancer efficacy and numerous
research efforts have attempted to generate more potent
noscapine analogues. These modifications included the replace-
ment of the N-methyl group in the 6’-position with a range of
substituents, where N-ethylcarbamoyl substitution was ob-
served to possess enhanced anticancer activity. Herein, we
describe advances in this area, namely the synthesis and
pharmacological evaluation of a series of N-sulfonyl and N-
sulfamoyl noscapine derivatives. A number of these sulfonyl-
containing noscapinoids demonstrated improved activities
compared to noscapine. ((R)-5-((S)-4,5-Dimethoxy-1,3-dihydroi-
sobenzofuran-1-yl)-4-methoxy-6-((1-methyl-1H-imidazol-4-yl)
sulfonyl)-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]isoquinoline) (14q)
displayed sub-micromolar activities of 560, 980, 271 and
443 nM against MCF-7, PANC-1, MDA-MB-435 and SK-MEL-5
cells, respectively. This antiproliferative effect was also main-
RES
tained against drug-resistant NCI/Adr
cells despite high
expression of the multidrug efflux pump, P-glycoprotein.
Introduction
Cancer is an enormous global health burden and is growing at
an alarming pace. As of 2018, cancer accounts for 1 in 6 deaths
[1]
worldwide. This figure is more than the combined death toll
from three other major public health problems: HIV/AIDS,
[2]
tuberculosis, and malaria. Chemotherapy remains an impor-
tant treatment modality in primary, adjuvant and palliative
settings. Despite continued improvements in anticancer drug
design over the last four decades, the most significant impair-
ment to chemotherapy is the emergence of an intrinsic or
[3,4]
acquired resistant phenotype.
Efforts to develop new anti-
cancer drugs must ensure potency, tumour selectivity, minimal
side effects and low susceptibility to resistance.
The importance of microtubules as a target for cancer
research has gained popularity since the clinical success of
vinca alkaloids, namely vincristine (1) and vinblastine (2) in the
[5]
[6]
1
960s, followed by paclitaxel (3) in the early 1990s (Figure 1).
Tubulin-binding agents (TBAs) that disrupt microtubule (MT)
dynamics produce cell cycle arrest, which triggers apoptosis
Figure 1. Antimitotic agents: vincristine (1), vinblastine (2) and paclitaxel (3).
[6–7]
through various mechanisms.
The current antimitotics can
be classified as tubulin-depolymerisation inhibitors (TDIs) (e.g.
paclitaxel) or tubulin-polymerisation inhibitors (TPIs) (e.g.
vincristine, vinblastine).
Tubulin targeting drugs are widely used in the clinic,
however they display several side effects including leukocyto-
[
8]
penia, alopecia, diarrhoea and peripheral neuropathies. There
are two well-established pathways of resistance towards MT
drugs : i) amplification of multi-drug resistant (MDR) protein,
[9]
specifically P-glycoprotein (P-gp) that leads to the efflux of drug
[8]
from tumour cells; and ii) high levels of expression of βIII-
tubulin subunits, an isotype that mediates its effect by
protecting cells against genotoxic stress induced by cytotoxic
[
a] C. Yong, Dr. S. M. Devine, Dr. B. Capuano, Prof. P. J. Scammells
Medicinal Chemistry, Monash Institute of Pharmaceutical Sciences
Monash University
[10]
drugs.
These dose-limiting toxicity issues and the gradual
381 Royal Parade, Parkville, VIC 3052 (Australia)
development of drug-resistant phenotypes has necessitated the
search for new tubulin-targeting chemotherapeutics.
E-mail: peter.scammells@monash.edu
b] X. Gao, A. Yan, Prof. R. Callaghan
[
Research School of Biology and Medical School
Australian National University
Canberra, ACT 0200 (Australia)
Noscapine (4), a phthalideisoquinoline alkaloid isolated
from the opium poppy, Papaver somniferum, has long been
used as an antitussive agent since the discovery of its cough-
Supporting information for this article is available on the WWW under
https://doi.org/10.1002/cmdc.201900477
[11]
suppressing effect in the 1950s (Figure 2). It was not until the
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We previously synthesised a series of N-substituted nosca-
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pine analogues as part of a SAR exploration at the 6’-position of
[18]
noscapine. Following the synthesis of a cyclic ether derivative
of N-nornoscapine, reaction with a range of alkyl halides, acid
chlorides, isocyanates, thiocyanates, and chloroformate re-
agents gave a library of noscapine derivatives: N-alkyl (5a), N-
acyl (5b), N-carbamoyl (5c), N-thiocarbamoyl (5d) and N-
alkoxycarbonyl (5e) analogues, respectively (Figure 2). Pharma-
cological evaluation of these derivatives was conducted against
prostate cancer (PC3), breast cancer (MCF-7) and colon cancer
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(
(
Caco-2) cell lines. Molecules with N-carbamoyl functionality
5c) were the most potent analogues, with N-ethylcarbamoyl
noscapine (6) exhibiting EC50 values of 3.6 μM and 6.7 μM,
against MCF-7 and PC3 cells, respectively. These results indicate
a preference for a hydrogen bonding motif at this region of
noscapine. Other modifications to the noscapine scaffold have
Figure 2. Noscapine (4) and N-substituted noscapine analogues (5a–e, 6).
[19–20]
been focused on the 1-, 7-, 6’- and 9’-positions.
changes included 7-demethylation
These
[21–23]
to O- and N-linked
noscapinoids, halogen insertion and formylation at the 9’-
[24À 25]
[26]
late 1990s, where noscapine was also found to possess weak
anticancer activity against cervical (HeLa), human breast (MCF-
position
and 6’-modifications.
In this study, we targeted N-sulfonyl and N-sulfamoyl
7
2
) and bladder (Renal 1983) cancer cell lines, with IC values of
5, 42 and 39 μM, respectively. Noscapine is a promising TPI,
noscapine analogues based on the promising data we obtained
5
0
[12]
[18]
on the N-substituted analogues described above.
The
and like other MT drugs, noscapine disrupts MT dynamics,
causing an alteration in MT conformation and assembly
sulfonamide functionality can be found in a diverse range of
approved medicines. Commonly found in antibacterial agents
[12–13]
[27]
properties.
Noscapine has demonstrated the ability to
(sulfamethoxazole, 7), sulfonamide moieties can also be seen
[
28]
cause G /M arrest and subsequent apoptosis in various cancer
in non-steroidal anti-inflammatory drugs (celecoxib, 8), and
2
[29]
cells. Immunofluorescence staining in HeLa cells following a 48-
hour treatment with noscapine demonstrated condensed
chromosomes, large amounts of fragmented nuclei and
diuretics (furosemide, 9)
(Figure 3). N-Substituted or N,N-
disubstituted sulfonamides are typically metabolically robust
groups that are not vulnerable to phase I or phase II
[12]
[30]
apoptotic morphologies.
metabolising enzymes. The isosteric replacement of carbonyl
As a range of established antimitotic agents including
vincristine (1), vinblastine (2), and paclitaxel (3), have proven
susceptible to the development of MDR, we studied the effect
of ABCB1 and ABCG2 transporters on selected noscapine
with the sulfonyl moiety not only retains hydrogen bonding
capabilities, but could also potentially reduce systemic clear-
ance of the molecule.
[14]
derivatives. The cell lines that were chosen for this study
WT
includes drug-sensitive MCF-7 parental cell line, ABCB1- Results and Discussion
RES
FLV1000
expressing (NCI-Adr ), and ABCG2-expressing (MCF-7
)
[15–16]
resistant breast cancer cells.
The results obtained were then
Chemistry
contrasted against the antimitotic agent, vinblastine (2) and
mitoxantrone, a type II topoisomerase inhibitor. A 260-fold and
The proposed synthesis of desired analogues follows a similar
[18]
1
7-fold decrease in antiproliferative potency was observed for
route previously reported. N-Demethylation of noscapine (4)
occurs via a non-classical Polonovski reaction to give N-
nornoscapine (10) (Scheme 1). The presence of the labile
lactone moiety in the southern heterocycle could be detrimen-
vinblastine and mitoxantrone in resistant cell lines, indicating
that these clinical agents are substrates for ABCB1 and ABCG2,
respectively. In contrast, noscapinoids that were evaluated
against the same drug-sensitive and resistant cell lines showed
negligible difference in antiproliferative potency. This indicates
that noscapine derivatives are unlikely to be substrates for
either ABCB1 or ABCG2. This discovery led to further studies
with purified, reconstituted P-gp in direct functional assays,
where the authors discovered that inhibition of P-gp function
[
17]
was due to direct interaction with the transporter.
These
enhanced qualities exhibited by the current noscapine deriva-
tives over existing antimitotic agents, as well as the oral
bioavailability and demonstrated safety profile make noscapine
an attractive molecule for further investigation.
Figure 3. Approved medications containing sulfonamide functionality (7–9).
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[
18]
Scheme 1. Reagents and conditions: a) 1. m-CPBA, CHCl
3
, 0°C, 2. HCl, 1 h, 95%, 2. FeSO4·7H
2
O, MeOH, À 5°C, 1 h, 78%; b) NaBH O, THF, 60°C, 4 h,
4
/BF3·Et
2
[
18]
1 2
3
2
8%; c) CSI, BnOH, Et
3
N, DCM, 0°C, 1 h, 29%; d) MeSO
2
Cl, Et
3
N, DCM, 25°C, 16 h, 63%; e) RÀ SO
2
Cl (14a–aa) or R R NÀ SO
Cl (15b–n), Et N or DBU, DCM,
2 3
5 C, 16 h, 14–85% (14a–aa) or 10–65% (15b–n); f) H
°
2
, Pd/C, AcOH, MeOH, 25 C, 16 h, 54%.
°
tal in vivo, and the demonstrated activity gains of cyclic ether
analogues, resulted in conversion of N-nornoscapine (10) to the
original reaction conditions were modified to assist progression
of the reaction: i) an increase in temperature from 25°C to
50°C; ii) an increase in reaction time from 16 h to 48 h; iii) an
increase in molar equivalence of the sulfonyl chloride reagent,
to no avail. Triethylamine was then replaced with 1,8-
diazabicycloundec-7-ene (DBU), a stronger non-nucleophilic
base, and this modification facilitated the progression of these
reactions. While there have been multiple derivatisation studies
conducted on noscapine (4), the inclusion of heteroaromatic
moieties has yet to be investigated. We anticipate the inclusion
of heteroaromatics 14m–r could be beneficial, in particular with
the inherent solubility issues that we have often observed with
noscapine in pharmacological studies. In general, the synthesis
of sulfonamides 14a–aa (yields ranging from 14–85%), were
better yielding than that of the sulfamides 15b–n (yields of 10–
65%). For example, 14a was afforded with a yield of 75%,
whereas the corresponding N-sulfamoyl 15b was obtained in
50% yield. Due to instability of the unsubstituted sulfamoyl
chloride reagent, synthesis of corresponding sulfamides, such
as 15a, often required an additional protection step (Scheme 1).
Carboxybenzyl (CBz)-protection of the aforementioned reagent
is achieved in situ through reaction of chlorosulfonyl isocyanate
cyclic ether 11 through
reduction.
a
BF .Et O/NaBH -mediated
3 2 4
Finally, reaction with the appropriate sulfonyl
[26,31]
chloride and sulfamoyl chloride afforded a diverse range of N-
sulfonyl (14a–aa) and N-sulfamoyl (15b–n) noscapine deriva-
tives, respectively. Methyl-substituted sulfonamide 13 was
synthesised from N-nornoscapine (10) in a similar fashion. Given
the replacement of N-methyl with N-ethylcarbamoyl (6) has
[18]
proved to be advantageous for anticancer activity,
the
corresponding sulfonamide (14b) and sulfamide (15c) ana-
logues, along with other alkyl chains (14a, 14c–g, 15b, 15d–g)
and cycloalkyls (14h–l, 15h–j), were synthesised to investigate
if the observed antiproliferation effects were maintained.
Further modifications included heterocyclic (14m–r, 15k–n),
aromatic (14s), benzyl (14t) and mono-substituted aromatics
(
14u–aa) to further assess potential activity gains.
[32]
Under the conditions prescribed by Park et al.,
the
synthesis of 14d (isopropyl), 14f (sec-butyl), 14i (cyclobutyl),
4j (cyclopentyl) and 14k (cyclohexyl) were not achieved. We
speculate nucleophilic substitution was prevented due to the
presence of a bulkier side chain behaving as a steric block. The
1
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(
CSI) with benzyl alcohol (BnOH). Following the addition of
7 cells halted in G /M to complete loss of activity. The G /M
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cyclic ether N-nornoscapine (11) under basic conditions, CBz-
protected sulfamide 12 is generated. The CBz-protecting group
is subsequently removed via hydrogenolysis using hydrogen
with palladium as catalyst, affording 15a in a yield of 54%.
arrest produced by unsubstituted phenyl (14s) was maintained
in the methoxy- (14w and 14x) and chloro-substituted phenyl
(14z and 14aa), with the exception of the ortho-substituted
analogues (14v and 14y).
Despite the presence of an additional nitrogen atom, G /M
2
activities for sulfamides (15a–n) were similar to those of the
sulfonamides (14a–aa). The sulfamide noscapinoids 15h and
Pharmacology
1
5k containing cyclopropyl and azetidine moieties, generated
an increase of MCF-7 cells trapped in mitosis by 303% and
06%, respectively. This indicated a preference for small cyclo-
Cell Cycle Arrest Assays
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N-Sulfonyl (13, 14a–aa) and N-sulfamoyl (15a–n) noscapine
derivatives were evaluated in cell cycle arrest assays based on
their ability to elicit G /M phase arrest through the disruption of
microtubule assembly. Human breast adenocarcinoma (MCF-7)
and human pancreatic cancer (PANC-1) cell lines were treated
with the test compound (10 μM) for 18 h. The extent of G /M
arrest (%) was calculated with reference to the untreated
control (0.05% v/v DMSO) and are presented in Table 1.
Pharmacological evaluation of N-sulfonyl and N-sulfamoyl
noscapinoids in cell cycle arrest assays showed promising
results, where 20 out of 41 derivatives displayed mitotic arrest
activities of greater than 200% in MCF-7 or PANC-1 cells.
Despite 13 being inactive in the MCF-7 cell line, transformation
of the lactone moiety to the cyclic ether (14a) led to a
pronounced increase in activity, with 267% MCF-7 cells arrested
in mitosis. This observation is consistent with previous findings,
where the removal of the lactone moiety was reported to
alkyl functionality within the sulfamide collection. Compound
14b showed the greatest G /M arrest activity in this series, with
2
322% more MCF-7 cells in mitotic arrest (Figure 4). This
observation is consistent with our earlier findings, where the
ethyl substituent from the N-carbamoyl series showed an
increase of 172% against MCF-7 cells and was the most active
2
2
[
18]
analogue from the exploration work at the N6’ position.
Potency (EC ) Determination
5
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Preliminary cell cycle arrest studies demonstrated that twenty
N-sulfonyl and N-sulfamoyl noscapine derivatives displayed
greater than 200% mitotic arrest in either one of the cell lines
(MCF-7 and PANC-1). The potency (EC ) of these sulfonamides
5
0
and sulfamides was determined with a fluorimetric cell viability
assay (CellTiter-Blue assay®). Results obtained from cell viability
studies were highly encouraging, where the majority of
analogues showed remarkable improvement from parent
noscapine (Table 1). Eleven of the tested analogues (14a, 14b,
14c, 14j, 14m, 14o, 14q, 14s, 14u, 15b and 15h) displayed
EC50 values of <2 μM in either MCF-7 or PANC-1 cells. The
thiophen-2-yl sulfonamide 14m displayed EC50 values of
1.47 μM and 4.76 μM against MCF-7 and PANC-1 cell lines,
respectively. This relatively small difference in activity indicates
that the efficacies of N-sulfonyl and N-sulfamoyl noscapine
derivatives are cell-line-dependant. More notably, 14q exhib-
ited sub-micromolar EC50 values of 560 nM against MCF-7 and
980 nM against PANC-1 cells; a 64- and 20-fold improvement
compared to noscapine (4). This observation suggests hetero-
aromatics with hydrogen bonding capabilities in this position
are favoured for improved tubulin-binding at this region of
noscapine.
[18,31]
promote antimitotic activity.
In general, the ability to produce G /M arrest activity was
2
not affected by the size of substituents. Similar mitotic activities
were observed for 14a–f, bearing simple alkyl side chains,
when contrasted against 14s–aa with larger aromatic substitu-
ents. From the small subset of alkyl substituents (14a–f), ethyl
(
14b) exhibited prominent activity (with 322% MCF-7 cells
arrested in G /M), followed by methyl (14a), n-propyl (14c), sec-
2
butyl (14f) and isopropyl (14d). With a slightly longer side
chain, the n-butyl (14e) led to a loss in activity, indicating the
preference for smaller alkyl chains. Replacement of the terminal
hydrogens in 14b with fluorine in 14g resulted in a reduction
in activity against both MCF-7 and PANC-1 cells. The activities
of cycloalkyl compounds 14h–k were relatively similar (~200%
against MCF-7 cells; ~150% against PANC-1 cells), with the
exception of cyclopropyl (14h). The 3,3-difluoro-substitution of
cyclobutyl (14l), also led to a complete loss of activity, when
compared with the non-substituted cyclobutyl analogue, 14i.
Sulfonamides with heteroaromatic functionalities (14m–r)
showed encouraging results. The activities obtained for thienyl
and furanyl-substituted analogues indicated a preference for
the 2-position (14m and 14o) over the 3-position (14n and
Resistance Profiles of the Sulfonyl-Containing Noscapinoids
With repeated and prolonged administration of current anti-
mitotic agents, drug resistance has become a common
observation. This is predominantly due to the amplification of
multi-drug resistant (MDR) proteins, specifically P-glycoprotein
(P-gp) that leads to efflux of drug from tumour cells,
1
4p). With an N-methylimidazol-4-yl side chain, 14q, displayed
profound antimitotic activity, with 246% and 166% cells
arrested in mitosis against MCF-7 and PANC-1 cells, respectively.
This is an outstanding 36-fold improvement when contrasted
with noscapine (4) in MCF-7 cells. A remarkable difference was
observed through the insertion of a methylene linker to the
phenyl substituent on 14s, giving benzyl 14t, from 229% MCF-
[
33]
subsequently reducing the therapeutic efficacies of the drug.
This effect can be observed in vinblastine (2), where the
potency decreased by 438-fold in the presence of P-gp in NCI/
RES
Adr cells (EC : 228�28 nM) compared to the drug-sensitive
5
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Table 1. Percentage increase in arrested cells trapped in the G
2
/M phase (tested at 10 μM) and EC50 values for N-sulfonyl and N-sulfamoyl noscapine.
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[
a]
[b]
Cpd
X
R
Change in %G
MCF-7
2
/M arrest
EC50 [μM]
MCF-7
PANC-1
PANC-1
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6�2
36�14
33�7
n3 5d .9�0.10
6.57�0.12
7.09�0.10
2.64�0.11
nd
n1 9d .3�0.12
1.52�0.09
1.26�0.07
0.46�0.08
nd
1
1
1
3
C=O
Me
Me
Et
À 1�2
4a
4b
CH
CH
CH
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CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
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CH
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CH
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2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
267�8
322�20
246�17
197�22
2�0.4
249�30
317�15
259�49
125�27
58�16
138�19
90�12
67�31
136�21
125�27
195�30
11�0.3
178�30
80�13
99�16
65�8
14c
n-Pr
i-Pr
n-Bu
s-Bu
14d
14e
14f
nd
nd
[c]
204�2.1
211�29
67�16
215�5
211�12
222�28
17�6
17.3�0.09
2.13�0.07
nd
5.35�0.09
1.13�0.04
9.14�0.16
nd
na
[
c]
14g
CH
2
CF
3
na
1
1
1
4h
4i
4j
cyclopropyl
cyclobutyl
cyclopentyl
cyclohexyl
3,3-difluorocyclobutyl
thiophen-2-yl
thiophen-3-yl
furan-2-yl
nd
[
c]
na
na
[c]
14k
n3 .d8 3�0.17
n4 .d7 6�0.12
n0 .d7 4�0.10
n0 .d9 8�0.04
n1 .d8 7�0.16
n0 .d9 9�0.12
nd
1
1
1
4l
4m
4n
284�24
139�7
203�11
83�1
n1 .d4 7�0.11
14o
n1 9d .9�0.08
n0 .d5 6�0.08
n4 .d7 7�0.10
n9 .d5 3�0.09
nd
14p
14q
14r
furan-3-yl
N-methylimidazol-4-yl
N-methylpyrazol-4-yl
Ph
Bn
4-MePh
2-OMePh
3-OMePh
4-OMePh
2-ClPh
3-ClPh
4-ClPh
NH
2
NHMe
NHEt
246�21
173�18
229�17
-5�7
166�27
59�15
132�19
30�5
14s
1
1
1
4t
4u
4v
244�12
66�6
210�26
43�17
135�33
278�52
71�30
304�35
109�34
73�17
58�23
48�17
49�19
33�23
46�14
16�21
247�43
17�27
37�3.3
209�14
49�19
91�10
41�15
14w
185�7.4
296�6
10�3.6
291�33
175�13
209�24
238�26
217�8
184�11
114�20
176�30
73�11
303�24
36�44
67�3.5
306�26
119�1
163�20
73�21
[
c]
[c]
14x
14y
14z
na
nd
na
nd
14aa
1n 3d .6�0.09
3.53�0.15
0.57�0.08
28.6�0.13
608�0.29
nd
6n .d6 9�0.13
149�0.35
6.11�0.09
39.1�0.17
15a
15b
15c
[
c]
15d
NEt
2
na
nd
1
1
1
5e
5f
5g
NH(n-Pr)
NH(i-Pr)
NH(i-Bu)
NH(cyclopropyl)
NH(cyclopentyl)
NH(cyclohexyl)
azetidin-1-yl
piperidin-1-yl
morpholin-4-yl
N-methylpiperazin-1-yl
[
c]
[c]
na
na
nd
nd
15h
n4 .d0 2�0.25
nd
4n 3d .5�0.13
n4 5d .3�0.13
n1 .d2 8�0.25
nd
6n .d6 6�0.22
440�0.39
nd
1
1
1
5i
5j
5k
15l
1
1
5m
5n
[
a]
2
Following FACS analysis of the treated cells, % increase in G /M arrest was calculated with reference to the vehicle control (0.05% DMSO). The reported
[b]
percentages represent the mean � SEM observed in three independent experiments. Cell viability was determined using standard resazurin reduction
[c]
method with CellTiter-Blue assay®. EC50 values shown represents the mean � SEM for at least 3 independent observations. EC50 values for compounds were
not attained due to solubility issues in the culture media
MCF-7 cells (EC : 0.52�0.05 nM) (Table 2). Consequently, NCI/
1.0 μM) in MCF-7 cells was markedly lower than that of
50
RES
Adr
cells are considered highly resistant. The continual
vinblastine and is in agreement with previous reports
[34]
evaluation of noscapinoids in resistance cancer cell lines is
critical to ensure resistance issues are mitigated. Noscapinoids
(Table 2).
The sulfonyl-containing noscapinoids displayed
fold-resistance (FR) values of 1.1 to 3.0 (Table 2). This indicates
the potencies observed in the drug-sensitive (MCF-7) were
1
4j, 14m, 14q and 15b displayed EC50 values of <1.5 μM
RES
against MCF-7 (refer to Supp. Info – Figure S1). These
compounds were further subjected to antiproliferative studies
against drug-sensitive (MCF-7) and drug-resistant (NCI/Adr
breast cancer cells. The potency of noscapine (4, EC : 11.9�
maintained against drug-resistant (NCI/Adr ) cells and demon-
strates the potential of these N-sulfonyl and N-sulfamoyl
compounds to evade common mechanisms of efflux.
RES
)
50
ChemMedChem 2019, 14, 1–15
www.chemmedchem.org
5
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
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These are not the final page numbers!

Full Papers
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2
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7
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9
0
1
2
3
4
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7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
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3
3
4
4
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4
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4
4
4
4
5
5
5
5
5
5
5
5
Figure 4. FACS analysis of MCF-7 cells treated for 18 h with (A) 0.05% DMSO, (B) vincristine (100 nM), (C) noscapine (4) (10 μM), (D) noscapine (4) (50 μM), (E)
ethyl sulfonamide (14b) (10 μM) and (F) N-methylimidazol-4-yl sulfonamide (14q) (10 μM). FACS data was processed with FlowJo (v10) and % cells were
obtained at the G (blue), S (yellow) and G /M (green) phases of the cell proliferation cycle.
1 2
Table 2. Efficacies of vinblastine (2), noscapine (4) and compounds 14j,
4m, 14q and 15b in drug-sensitive (MCF-7) and drug-resistant (NCI/
ADR ) breast cancer cell lines.
1
determine if the antimitotic effect of 14q is due to interaction
with tubulin subunits, tubulin polymerisation studies were done
in the absence or presence of known tubulin ligands (10 μM) –
RES
[a]
Cpd
EC50 [μM]
RES
[c]
MCF-7
NCI/Adr
FR
paclitaxel (3) and noscapine (4) (Figure 5). The control (Vmax
=
[b]
2
0.52�0.05
11.9�1.0
228�28
438
2.1
1.1
3.0
2.2
2.4
2
4.52�1.23 mOD/min) represents a standard polymerisation
4
24.6�1.70
2.15�0.40
4.65�1.01
1.39�0.26
3.48�0.69
curve observed at 37°C over a period of 60 minutes. Paclitaxel
(3), a microtubule-stabilising agent, was found to enhance the
Vmax by approximately three-fold (Vmax =70.67�6.19 mOD/min).
1
1
1
4j
4m
4q
1.93�0.44
1.54�0.34
0.62�0.09
1.47�0.20
15b
[12]
Noscapine (4), similar to previous findings, led to a reduction
of the polymerisation rate (Vmax =18.05�0.98 mOD/min). At
[a]
The potency (EC50) of noscapinoids to cause antiproliferative effects was
estimated from non-linear least squares regression of the general dose-
1
0 μM, compound 14q significantly reduced the initial polymer-
response relationship. All values correspond to the mean�SEM obtained
[
b]
from at least three independent observations.
The EC50 values for
isation rate to 3.241�0.048 mOD/min. Moreover, 14q remained
in the growth phase of the polymerisation curve throughout
the duration of the experiment, whilst the other tubulin ligands
[c]
vinblastine are represented in nM. The fold-resistance (FR) was calculated
as the ratio of EC50 values between the two cell lines.
(paclitaxel and noscapine) reached a steady state at t=20
onwards.
Inhibition of Tubulin Polymerisation by 14q
The antiproliferative effect of antitubulin agents are typically
due to interference with MT dynamics, through either inhibition
of polymerisation or depolymerisation of tubulin subunits.
NCI-60 Human Tumour Cell Lines Screen
[35]
Given the positive antimitotic discoveries with noscapinoid 14q
thus far, we further explored the anticancer potential of 14q
against different cancer types. Testing was performed by the
Developmental Therapeutics Program, Division of Cancer Treat-
Tubulin-binding studies have previously discovered that nosca-
pine reduces the rate of polymerisation, represented as
maximal initial velocity (Vmax), in a concentration-dependant
[
12]
manner.
This microtubule-destabilising effect of noscapine
ment and Diagnosis, National Cancer Institute (http://dtp.can-
[23,34,36]
[37],[38]
has also been retained by several noscapinoids.
To
cer.gov).
Compound 14q was evaluated against a panel of
ChemMedChem 2019, 14, 1–15
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6
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
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fold. The findings relating to 14q are noteworthy as, to the best
of our knowledge, noscapine derivatives that have been
reported to date are predominantly in the low micromolar
range.
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
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3
4
5
6
7
8
9
0
1
2
3
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6
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8
9
0
1
2
3
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9
0
1
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7
Experimental Section
Chemistry
1
19
13
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
H, F and C NMR spectra were recorded on a Bruker Avance
Nanobay III 400 MHz Ultrashield Plus spectrometer at 400.13,
76.85 and 100.62 MHz, respectively coupled to a BACS 60
3
automatic sample changer at 25°C. Chemical shifts (δ) are recorded
in parts per million (ppm) by correction with reference to the
chemical shift of the solvent, according to the procedure described
Figure 5. Tubulin-polymerisation in the absence (*) and presence of tubulin
ligands, paclitaxel ( ), noscapine (&) and 14q (~) at 10 μM. Paclitaxel, as a
TDI, resulted in an increased Vmax by approximately three-fold
◆
[23]
by Gottlieb. Coupling constants (J) are recorded in Hz, and the
(
V
max =70.67�6.19 mOD/min). Noscapine and 14q exhibited characteristic
significant multiplicities described by singlet (s), doublet (d), triplet
TPI effect on the rate of tubulin polymerisation (Vmax =18.05�0.98 mOD/min
(
(
t), quadruplet (q), broad (br), multiplet (m), doublet of doublets
dd), and doublet of triplets (dt). LC-MS were run to verify reaction
and 3.241�0.048 mOD/min, respectively).
outcome and purity using an Agilent 6120 series Single Quad
coupled to an Agilent 1260 series HPLC. The following buffers were
used: buffer A, 0.1% formic acid in H O; buffer B, 0.1% formic acid
2
6
0 cancer cells, which holds cell lines representing leukemia,
in MeCN. The following gradient was used with a Phenomenex
Luna 3 μM C8(2) 15 mm ×4.6 mm column, and a flow rate of
non-small cell lung carcinoma, melanoma and cancers of the
colon, brain, ovary, kidney, prostate and breast. Compound 14q
displayed significant growth inhibition activities of <500 nM
against five different cancer types: leukemia (K-562), melanoma
0
.5 mL/min and total run time of 12 min; 0–4 min 95% buffer A and
5% buffer B, 4–7 min 0% buffer A and 100% buffer B, 7–12 min
5% buffer A and 5% buffer B. Mass spectra were acquired in
9
positive and negative ion mode with a scan range of 0–1000 m/z at
5 V. UV detection was carried out at 214 nm and 254 nm. All
screening compounds were of >95% purity. Thin layer chromatog-
raphy was conducted on 0.2 mm plates using Merck silica gel 60
F254. Column chromatography was achieved using Merck silica gel
60 (particle size 0.063–0.200 μm, 70–230 mesh).
(MDA-MB-435 and SK-MEL-5), non-small cell lung (NCI-H226 &
NCI-H522), colon (HCT-116), renal (RXF) and breast (MCF-7 and
MDA-MB-468) cancers (refer to Supp. Info – Figure S2 and S3).
Through this screening procedure, we have identified 14q as a
potent tumour growth inhibitor for melanoma cells, with GI50
values of 271 nM and 443 nM against MDA-MB-435 and SK-
MEL-5 cells, respectively. More notably, 14q also displayed
significant antiproliferative activity against SK-MEL-5 with an
LC50 of 38.6 μM.
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
[18]
thoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]isoquinoline (11). To
a cooled (À 5°C) solution of NaBH (219 mg, 5.78 mmol, 3 eq.) in
4
THF (7 mL) was added a solution of N-nornoscapine (10, 770 mg,
1
.93 mmol) in BF ·Et O (2.38 mL, 19.3 mmol, 10 eq.) dropwise. The
3 2
mixture was allowed to warm to 25°C for 1 h and then heated at
reflux for 2 h. The reaction was recharged with NaBH (219 mg,
4
Conclusions
5
.78 mmol, 3 eq.) and BF ·Et O (2.38 mL, 19.3 mmol, 10 eq.) and
3 2
heated at reflux for another 2 h. The reaction was then quenched
with the dropwise addition of cold 10% HCl (~20 mL) and allowed
In summary, a library of 42 N-sulfonyl and N-sulfamoyl nosca-
pine derivatives (13, 14a–aa and 15a–n) was synthesised.
These derivatives were readily accessed through reaction of
cyclic ether nor-noscapine (11) with the corresponding sulfonyl
or sulfamoyl chlorides. The vast majority of N-sulfonyl nosca-
pine derivatives displayed greater ability to promote mitotic
arrest compared to noscapine (4). Twenty of these compounds
to stir for 30 min before being extracted with CHCl (3×25 mL). The
3
organic layer was then washed with 10% NaOH (2×20 mL) to yield
the free base form. The organic layer was dried over MgSO and
4
evaporated under reduced pressure. The crude product was
purified with column chromatography (CHCl /MeOH/NH ; 85:14:1)
3
3
1
to afford 7 (286 mg, 38%) as a colourless foam. H NMR (CDCl ): δ
3
6.60 (d, J=8.2 Hz, 1H), 6.33 (s, 1H), 5.93 (d, J=1.4 Hz, 1H), 5.92 (d,
J=1.4 Hz, 1H), 5.84 (d, J=8.2 Hz, 1H), 5.77 (s, 1H), 5.36 (dd, J=12.3,
2.8 Hz, 1H), 5.18 (d, J=12.2 Hz, 1H), 4.62 (d, J=4.0 Hz, 1H), 4.00 (s,
3H), 3.85 (s, 3H), 3.80 (s, 3H), 2.70–2.52 (m, 3H), 2.38–2.23 (m, 1H).
displayed G /M arrest activity of >200% and were further
2
evaluated in cell viability studies, whereby 14j, 14m, 14q and
1
5b demonstrated anticancer activities of <1.5 μM against
(
S)-6,7-Dimethoxy-3-((R)-4-methoxy-6-(methylsulfonyl)-5,6,7,8-
MCF-7 cells. Noscapinoid 14q, bearing an N-methylimidazol-4-yl
substituent, possesses sub-micromolar activity (EC50 values:
tetrahydro[1,3]dioxolo[4,5-g]isoquinolin-5-yl)isobenzofuran-
1(3H)-one (13). To a solution of 10 (100 mg, 0.25 mmol, 1 eq.)
5
2
60 nM against MCF-7 cells; 980 nM against PANC-1 cells;
71 nM against MDA-MB-435 cells; 443 nM against SK-MEL-5
dissolved in DCM (1 mL) was added Et N (105 μL, 0.75 mmol, 3 eq.),
3
and the mixture was left to stir at 25°C for 15 min. Then,
cells). Unlike many clinical anticancer agents, this potency is
methanesulfonyl chloride (21 μL, 0.28 mmol, 1.1 eq.) was added
RES
°
C for 16 h. Water (~5 mL) was
also maintained against drug-resistant NCI/Adr cells, evidence
and the reaction was stirred at 25
added to the reaction, and the solution was extracted with DCM
of circumvention of common efflux mechanisms. Furthermore,
(3×10 mL), dried with MgSO , filtered and filtrate evaporated under
4
1
4q demonstrated a strong MT destabilising profile, where the
reduced pressure to yield the crude product. The crude was purified
rate of tubulin polymerisation was significantly reduced by 8-
ChemMedChem 2019, 14, 1–15
www.chemmedchem.org
7
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
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These are not the final page numbers!

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+
by flash chromatography (1:1; EtOAc: pet. spirits) to afford 13 as
13.1. HR-ESMS calcd. for C H NO S [M+H] 492.1687, found
492.1688.
2
4
30
8
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
1
pale yellow foam (75 mg, 63%). H NMR (CDCl ): δ 7.03 (d, J=
3
8
.3 Hz, 1H), 6.34 (s, 1H), 6.31 (dd, J=8.2, 0.8 Hz, 1H), 5.97–5.92 (m,
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-6-(iso-
3H), 5.71 (d, J=4.5 Hz, 1H), 4.07 (s, 3H), 4.01 (s, 3H), 3.87 (s, 3H),
3.55–3.46 (m, 1H), 3.00 (s, 3H), 2.79 (ddd, J=17.5, 8.8, 4.1 Hz, 1H),
propylsulfonyl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]
isoquinoline (14d). Compound 14d was synthesised from 11
(99 mg, 0.26 mmol, 1 eq.) according to General Procedure A, with
DBU (192 μL, 1.28 mmol, 5 eq.) and 2-propanesulfonyl chloride
(64 μL, 0.57 mmol, 2.2 eq.) in DCM at 50
was obtained as a colourless oil (75 mg, 59%). H NMR (CDCl ): δ
2
.44 (ddd, J=16.6, 4.1, 2.8 Hz, 1H), 2.19 (ddd, J=14.0, 11.2, 4.2 Hz,
13
1H). C NMR (CDCl ): δ 167.7, 152.9, 149.4, 148.5, 139.9, 139.9,
3
1
34.4, 129.6, 118.9, 118.8, 118.2, 114.7, 103.3, 101.2, 79.4, 62.6, 59.8,
+
°
C for 16 h. The product
56.9, 53.2, 40.6, 40.1, 28.0. HR-ESMS calcd. for C H NO S [M+H]
22
24
9
1
3
478.1166, found 478.1171.
6
.69 (d, J=8.2 Hz, 1H), 6.34 (s, 1H), 6.25 (d, J=8.2 Hz, 1H), 5.90 (s,
General Procedure A: Sulfonamide or Sulfamide Synthesis. To a
solution of 11 (typically 100 mg, 1.0 eq.) dissolved in DCM or DMF
2H), 5.64 (s, 1H), 5.40 (d, J=4.1 Hz, 1H), 5.15–5.03 (m, 2H), 3.85 (s,
3H), 3.82 (s, 3H), 3.82 (s, 3H), 3.51 (dd, J=17.3, 10.2 Hz, 1H), 3.33 (dt,
J=13.6, 6.8 Hz, 1H), 2.90–2.73 (m, 2H), 2.61–2.49 (m, 1H), 1.35 (d,
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
(1 mL) was added the Et N or DBU (typically 3.0 eq.), and the
3
1
3
mixture was left to stir at 25°C for 15 min. The appropriate sulfonyl
or sulfamoyl chloride (typically 1.5 eq.) was added and the reaction
was stirred at the described conditions. Water (~5 mL) was added
to the reaction, the solution extracted with DCM (3×10 mL), dried
J=6.9 Hz, 3H), 1.20 (d, J=6.8 Hz, 3H). C NMR (CDCl ): δ 151.6,
148.7, 143.2, 139.9, 134.2, 132.9, 132.2, 129.4, 117.9, 117.2, 112.3,
103.0, 100.9, 85.5, 71.6, 60.1, 59.4, 56.4, 55.6, 53.8, 40.6, 28.1, 16.9,
16.7. HR-ESMS calcd. for C H NO S [M+H] 492.1687, found
3
+
2
4
30
8
with MgSO , filtered and the filtrate evaporated under reduced
492.1692.
4
pressure to yield the crude product. The crude residue was purified
by flash chromatography (1:1; EtOAc: pet. spirits) to afford the
desired sulfonamides (14a–aa) and sulfamides (15a–n).
(
R)-6-(Butylsulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihydroisobenzofur-
an-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]isoquino-
line (14e). Compound 14e was synthesised from 11 (99 mg,
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
0.26 mmol, 1 eq.) according to General Procedure A, with Et N
(108 μL, 0.77 mmol, 3 eq.) and 1-butanesulfonyl chloride (50 μL,
3
thoxy-6-(methylsulfonyl)-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]iso-
quinoline (14a). Compound 14a was synthesised from 11 (99 mg,
0.39 mmol, 1.5 eq.) in DCM at 25°C for 16 h. The product was
1
0.26 mmol, 1 eq.) according to General Procedure A, with Et
N
3
obtained as a colourless oil (85 mg, 66%). H NMR (CDCl
): δ 6.68 (d,
3
(
108 μL, 0.77 mmol, 3 eq.) and methanesulfonyl chloride (30 μL,
.39 mmol, 1.5 eq.) in DCM at 25°C for 16 h. The product was
J=8.2 Hz, 1H), 6.32 (s, 1H), 6.22 (d, J=8.2 Hz, 1H), 5.96–5.82 (m, 2H),
5.68–5.57 (m, 1H), 5.37 (d, J=4.3 Hz, 1H), 5.09 (d, J=1.7 Hz, 2H),
3.87 (s, 3H), 3.81 (s, 6H), 3.62–3.43 (m, 1H), 3.08–2.86 (m, 2H), 2.82–
2.67 (m, 2H), 2.61–2.45 (m, 1H), 1.85–1.55 (m, 2H), 1.47–1.28 (m, 2H),
0
1
obtained as a colourless oil (90 mg, 75%). H NMR (CDCl ): δ 6.68 (d,
3
J=8.2 Hz, 1H), 6.32 (s, 1H), 6.23 (d, J=8.2 Hz, 1H), 5.89 (s, 2H), 5.65
1
3
(d, J=4.2 Hz, 1H), 5.37 (d, J=4.3 Hz, 1H), 5.09 (d, J=1.4 Hz, 2H),
0.88 (t, J=7.4 Hz, 3H). C NMR (CDCl ): δ 151.5, 148.7, 143.1, 139.9,
134.1, 132.7, 132.2, 129.3, 117.7, 116.8, 112.2, 102.9, 100.8, 85.2,
71.7, 60.1, 59.3, 56.3, 55.3, 52.1, 40.0, 27.8, 25.4, 21.7, 13.7. HR-ESMS
3
3.88 (s, 3H), 3.81 (s, 6H), 3.59–3.50 (m, 1H), 2.84 (s, 3H), 2.79–2.68 (m,
13
2
1
7
H), 2.60–2.50 (m, 1H). C NMR (CDCl ): δ 151.6, 148.9, 143.1, 140.0,
34.1, 132.5, 132.2, 129.3, 117.7, 116.4, 112.2, 102.9, 100.9, 85.1,
1.8, 60.1, 59.4, 56.3, 55.4, 39.9, 39.2, 27.5. HR-ESMS calcd. for
3
+
calcd. for C H NO S [M+H] 506.1843, found 506.1843.
2
5
32
8
+
(5R)-6-(sec-Butylsulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihydroisoben-
zofuran-1-yl)-4-methoxy-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]iso-
quinoline (14f). Compound 14f was synthesised from 11 (85 mg,
0.22 mmol, 1 eq.) according to General Procedure A, with DBU
(147 μL, 0.99 mmol, 4.5 eq.) and 2-butanesulfonyl chloride (59 μL,
C H NO S [M+H] 464.1374, found 464.1372.
2
2
26
8
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-6-(ethyl-
sulfonyl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]isoqui-
noline (14b). Compound 14b was synthesised from 11 (99 mg,
0
.26 mmol, 1 eq.) according to General Procedure A, with Et N
0.46 mmol, 2.1 eq.) in DCM at 50°C for 24 h. The product was
3
1
(108 μL, 0.77 mmol, 3 eq.) and ethanesulfonyl chloride (37 μL,
obtained as a white foam (29 mg, 26%). H NMR (CDCl ) (rotamers:
3
#
0.39 mmol, 1.5 eq.) in DCM at 25°C for 16 h. The product was
0.4 :0.6*): δ 6.69 (d, J=8.2 Hz, 1H), 6.34 (s, 1H), 6.27–6.18 (m, 1H),
1
obtained as a yellow oil (87 mg, 71%). H NMR (CDCl ): δ 6.69 (d,
5.94–5.85 (m, 2H), 5.65 (s, 1H), 5.40 (dd, J=9.8, 4.1 Hz, 1H), 5.12–
5.06 (m, 2H), 3.85 (s, 3H), 3.82 (s, 6H), 3.57–3.45 (m, 1H), 3.09 (ddd,
J=10.1, 7.8, 5.0 Hz, 1H), 2.88–2.72 (m, 2H), 2.54 (ddd, J=11.4, 8.3,
3
J=8.2 Hz, 1H), 6.34 (s, 1H), 6.24 (d, J=8.1 Hz, 1H), 5.91 (s, 2H), 5.72–
5
3
2
.57 (m, 1H), 5.39 (d, J=4.1 Hz, 1H), 5.24–5.05 (m, 2H), 3.89 (s, 3H),
.83 (s, 6H), 3.61–3.45 (m, 1H), 3.11–2.93 (m, 2H), 2.84–2.70 (m, 2H),
.64–2.45 (m, 1H), 1.28 (t, J=7.4 Hz, 3H). C NMR (CDCl ): δ 151.6,
#
#
3.7 Hz, 1H), 2.12–1.75 * (m, 1H), 1.63–1.41 * (m, 1H), 1.33* (d, J=
# #
13
6.9 Hz, 2H), 1.18 (d, J=6.8 Hz, 1H), 1.00 (t, J=7.5 Hz, 1H), 0.95* (t,
3
1
3
148.8, 143.2, 140.0, 134.2, 132.8, 132.2, 129.4, 117.8, 116.9, 112.2,
103.0, 100.9, 85.2, 71.8, 60.2, 59.4, 56.4, 55.4, 46.8, 40.2, 27.8, 8.2.
HR-ESMS calcd. for C H NO S [M+H] 464.1374, found 464.1372.
J=7.5 Hz, 2H). C NMR (CDCl ): δ 151.5, 148.7, 143.1, 139.9, 134.2,
132.9, 132.2, 129.4, 117.8, 117.2, 112.3, 103.0, 100.9, 85.4, 71.6, 60.1,
59.7, 59.4, 56.4, 55.6, 40.5, 28.1, 23.5, 13.3, 11.2. HR-ESMS calcd. for
3
+
23 28
8
+
C H NO S [M+H] 506.1843, found 506.1848.
2
5
32
8
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-(propylsulfonyl)-5,6,7,8-tetrahydro-[1,3]dioxolo[4,5-g]iso-
quinoline (14c). Compound 14c was synthesised from 11 (99 mg,
(R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-((2,2,2-trifluoroethyl)sulfonyl)-5,6,7,8-tetrahydro[1,3]di-
oxolo[4,5-g]isoquinoline (14g). Compound 14g was synthesised
from 11 (99 mg, 0.26 mmol, 1 eq.) according to General Procedure
A, with DBU (107 μL, 0.77 mmol, 3 eq.) and 2,2,2-trifluoroethanesul-
fonyl chloride (43 μL, 0.38 mmol, 1.5 eq.) in DCM at 25°C for 16 h.
0
.26 mmol, 1 eq.) according to General Procedure A, with Et N
3
(108 μL, 0.77 mmol, 3 eq.) and 1-propanesulfonyl chloride (43 μL,
0
.39 mmol, 1.5 eq.) in DCM at 25°C for 16 h. The product was
1
obtained as a light brown oil (108 mg, 85%). H NMR (CDCl ): δ 6.67
(d, J=8.2 Hz, 1H), 6.31 (s, 1H), 6.20 (d, J=8.2 Hz, 1H), 5.89 (d, J=
1.4 Hz, 1H), 5.88 (d, J=1.4 Hz, 1H), 5.68–5.57 (m, 1H), 5.36 (d, J=
3
1
The product was obtained as a colourless oil (58 mg, 43%). H NMR
(CDCl ): δ 6.68–6.61 (m, 1H), 6.35 (s, 1H), 5.99 (d, J=8.2 Hz, 1H),
5.95–5.91 (m, 2H), 5.75–5.69 (m, 1H), 5.53 (d, J=4.4 Hz, 1H), 5.18–
5.03 (m, 2H), 4.18–4.04 (m, 1H), 3.97 (s, 3H), 3.94–3.83 (m, 1H), 3.83
(s, 3H), 3.81 (s, 3H), 3.62 (dd, J=13.3, 6.2 Hz, 1H), 2.87–2.75 (m, 1H),
2.73–2.62 (m, 1H), 2.55–2.45 (m, 1H). C NMR (CDCl ): δ 151.8,
149.0, 143.3, 139.8, 134.4, 132.2, 132.0, 129.0, 122.1 (q, J=277.4 Hz),
117.7, 115.9, 112.4, 103.0, 101.1, 84.3, 71.2, 60.2, 59.6, 56.3, 55.2,
3
4
3
1
1
1
.3 Hz, 1H), 5.09 (d, J=1.6 Hz, 2H), 3.87 (s, 3H), 3.80 (s, 6H), 3.56–
.44 (m, 1H), 3.04–2.84 (m, 2H), 2.80–2.65 (m, 2H), 2.57–2.46 (m, 1H),
.87–1.63 (m, 2H), 0.98 (t, J=7.5 Hz, 3H). C NMR (CDCl ): δ 151.5,
48.7, 143.0, 139.8, 134.1, 132.6, 132.1, 129.3, 117.7, 116.7, 112.2,
02.9, 100.8, 85.1, 71.6, 60.0, 59.3, 56.3, 55.2, 54.1, 40.0, 27.8, 17.1,
13
3
13
3
ChemMedChem 2019, 14, 1–15
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8
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
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1
9
13
5
4.6 (q, J=31.1 Hz), 40.3, 28.2. F NMR (377 MHz, CDCl ) δ À 62.00.
1.57–1.41 (m, 2H), 1.33–1.10 (m, 2H). C NMR (CDCl ): δ 151.5,
3
3
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
+
HR-ESMS calcd. for C H F NO S
[M+H] 532.1247, found
148.7, 143.2, 139.9, 134.2, 132.9, 132.2, 129.5, 117.9, 117.2, 112.3,
103.0, 100.9, 85.4, 71.5, 61.9, 60.1, 59.4, 56.4, 55.4, 40.6, 28.3, 26.6,
26.5, 25.5, 25.4, 25.4. HR-ESMS calcd. for C H NO S [M+H]
23
25
3
8
5
32.1248.
+
2
7
34
8
(
R)-6-(Cyclopropylsulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihydroiso-
532.2000, found 532.2001.
benzofuran-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]
isoquinoline (14h). Compound 14h was synthesised from 11
(107 mg, 0.28 mmol, 1 eq.) according to General Procedure A, with
(R)-6-((3,3-Difluorocyclobutyl)sulfonyl)-5-((S)-4,5-dimethoxy-1,3-
dihydroisobenzofuran-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]di-
oxolo[4,5-g]isoquinoline (14l). Compound 14l was synthesised
from 11 (94 mg, 0.24 mmol, 1 eq.) according to General Procedure
A, with Et N (102 μL, 0.73 mmol, 3 eq.) and (3,3-dimethylcyclobutyl)
methanesulfonyl chloride (75 mg, 0.37 mmol, 1.5 eq.) in DCM at
Et N (116 μL, 0.83 mmol, 3 eq.) and cyclopropanesulfonyl chloride
3
(
51 μL, 0.42 mmol, 1.5 eq.) in DCM at 50°C for 16 h. The product
1
was obtained as a white foam (58 mg, 42%). H NMR (CDCl ): δ 6.70
3
3
(
d, J=8.2 Hz, 1H), 6.35 (d, J=8.2 Hz, 1H), 6.32 (s, 1H), 5.89 (d, J=
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
1
.4 Hz, 1H), 5.88 (d, J=1.4 Hz, 1H), 5.62–5.56 (m, 1H), 5.38 (d, J=
25°C for 16 h. The product was obtained as a white foam (97 mg,
1
4.3 Hz, 1H), 5.16–5.04 (m, 2H), 3.84 (s, 3H), 3.81 (s, 3H), 3.80 (s, 3H),
72%). H NMR (CDCl ): δ 6.69 (d, J=8.2 Hz, 1H), 6.34 (s, 1H), 6.20 (d,
3
3.66–3.52 (m, 1H), 2.96–2.73 (m, 2H), 2.64–2.50 (m, 1H), 2.22 (tt, J=
8.0, 4.9 Hz, 1H), 1.21–1.02 (m, 2H), 0.92–0.67 (m, 2H). C NMR
J=8.2 Hz, 1H), 5.92 (s, 2H), 5.70–5.61 (m, 1H), 5.37 (d, J=4.3 Hz, 1H),
5.14–5.01 (m, 2H), 3.90 (s, 3H), 3.83 (s, 3H), 3.83 (s, 3H), 3.59–3.47 (m,
1H), 3.21 (dd, J=13.6, 8.2 Hz, 1H), 3.09 (dd, J=13.7, 6.7 Hz, 1H),
1
3
(
1
CDCl ): δ 151.5, 148.7, 143.0, 140.0, 134.0, 132.9, 132.2, 129.2, 117.7,
17.0, 112.1, 102.8, 100.8, 85.3, 71.8, 60.0, 59.3, 56.3, 55.7, 40.2, 29.6,
3
1
3
2.86–2.69 (m, 4H), 2.64–2.53 (m, 2H), 2.53–2.31 (m, 2H). C NMR
+
27.4, 5.4, 5.1. HR-ESMS calcd. for C H NO S [M+H] 490.1530,
(CDCl ): δ 151.7, 148.9, 143.2, 139.9, 134.3, 132.5, 132.2, 129.3, 121.3
24
28
8
3
found 490.1534.
(d, J=274.6 Hz), 117.8, 116.5, 112.4, 103.0, 101.0, 85.1, 71.7, 60.2,
5
9.5, 56.6, 56.4, 55.3, 41.0 (d, J=22.3 Hz), 40.6 (d, J=22.4 Hz), 40.1,
(
R)-6-(Cyclobutylsulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihydroiso-
19
27.9, 18.6. F NMR (377 MHz, CDCl
) δ À 83.31 (d, J=195.1 Hz),
3
benzofuran-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]
isoquinoline (14i). Compound 14i was synthesised from 11
+
À 95.27 (d, J=195.2 Hz). HR-ESMS calcd. for C H F NO S [M+H]
26 30
2
8
5
54.1655, found 554.1661.
(
89 mg, 0.23 mmol, 1 eq.) according to General Procedure A, with
DBU (155 μL, 1.04 mmol, 4.5 eq.) and cyclobutanesulfonyl chloride
(R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-(thiophen-2-ylsulfonyl)-5,6,7,8-tetrahydro[1,3]dioxolo
[4,5-g]isoquinoline (14m). Compound 14m was synthesised from
11 (83 mg, 0.21 mmol, 1 eq.) according to General Procedure A,
(54 μL, 0.49 mmol, 2.1 eq.) in DCM at 50
was obtained as a colourless oil (17 mg, 15%) H NMR (CDCl ): δ
°
C for 72 h. The product
1
3
6
.71 (d, J=8.2 Hz, 1H), 6.34 (d, J=8.2 Hz, 1H), 6.32 (s, 1H), 5.91 (d,
J=1.4 Hz, 1H), 5.89 (d, J=1.4 Hz, 1H), 5.58 (dd, J=3.0, 1.3 Hz, 1H),
with Et N (90 μL, 0.64 mmol, 3 eq.) and 2-thiophenesulfonyl
3
5
.34–5.28 (m, 1H), 5.09 (d, J=1.5 Hz, 2H), 3.84 (s, 3H), 3.83 (s, 3H),
.82 (s, 3H), 3.80–3.69 (m, 1H), 3.60–3.52 (m, 1H), 2.90–2.69 (m, 2H),
chloride (41 mg, 0.23 mmol, 1.05 eq.) in DCM at 25°C for 16 h. The
1
3
product was obtained as a colourless oil (92 mg, 81%). H NMR
2.59–2.48 (m, 2H), 2.41 (dq, J=17.7, 9.0 Hz, 1H), 2.25–2.13 (m, 1H),
(CDCl ): δ 7.46 (dd, J=3.7, 1.3 Hz, 1H), 7.43 (dd, J=5.0, 1.3 Hz, 1H),
3
13
2
1
7
.05–1.85 (m, 3H). C NMR (CDCl ): δ 151.6, 148.7, 143.1, 140.0,
6.95 (dd, J=5.0, 3.7 Hz, 1H), 6.73 (d, J=8.2 Hz, 1H), 6.48 (d, J=
8.2 Hz, 1H), 6.19 (s, 1H), 5.87 (d, J=1.4 Hz, 1H), 5.84 (d, J=1.4 Hz,
1H), 5.63–5.58 (m, 1H), 5.54 (d, J=4.1 Hz, 1H), 5.03 (d, J=12.3 Hz,
1H), 4.90 (dd, J=12.4, 2.7 Hz, 1H), 3.83 (s, 3H), 3.79 (s, 3H), 3.79 (s,
3
34.1, 133.0, 132.2, 129.2, 117.8, 117.3, 112.2, 102.9, 100.9, 85.4,
2.0, 60.1, 59.3, 56.4, 55.6, 54.3, 40.1, 27.6, 24.5, 24.4, 17.2. HR-ESMS
+
calcd. for C H NO S [M+H] 504.1687, found 504.1690.
25
30
8
3
H), 3.47–3.37 (m, 1H), 3.10–2.98 (m, 1H), 2.60 (dt, J=16.3, 5.3 Hz,
(
R)-6-(Cyclopentylsulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihydroiso-
13
1H), 2.50–2.40 (m, 1H). C NMR (CDCl ): δ 151.6, 148.6, 143.1, 140.7,
3
benzofuran-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]
isoquinoline (14j). Compound 14j was synthesised from 11
140.1, 134.0, 132.8, 132.3, 131.8, 131.5, 129.6, 127.1, 117.8, 116.4,
1
12.2, 102.5, 100.8, 86.1, 71.9, 60.1, 59.3, 56.4, 56.1, 41.0, 26.9. HR-
(
92 mg, 0.24 mmol, 1 eq.) according to General Procedure A, with
+
ESMS calcd. for C H NO S [M+H] 532.1094, found 532.1095.
2
5
26
8 2
Et N (161 μL, 1.08 mmol, 4.5 eq.) and cyclopentanesulfonyl chloride
3
(66 μL, 0.50 mmol, 2.1 eq.) in DCM at 50°C for 72 h. The product
(R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-(thiophen-3-ylsulfonyl)-5,6,7,8-tetrahydro[1,3]dioxolo
[4,5-g]isoquinoline (14n). Compound 14n was synthesised from
11 (51 mg, 0.13 mmol, 1 eq.) according to General Procedure A,
1
was obtained as a yellow oil (21 mg, 17%). H NMR (CDCl ): δ 6.70
3
(
d, J=8.2 Hz, 1H), 6.33 (s, 1H), 6.30 (d, J=8.2 Hz, 1H), 5.91 (d, J=
1
4
3
.1 Hz, 1H), 5.90 (d, J=1.3 Hz, 1H), 5.64–5.59 (m, 1H), 5.39 (d, J=
.3 Hz, 1H), 5.15–5.05 (m, 2H), 3.85 (s, 3H), 3.82 (s, 3H), 3.82 (s, 3H),
.62–3.49 (m, 2H), 2.91–2.74 (m, 2H), 2.55 (dd, J=12.1, 3.4 Hz, 1H),
with Et N (55 μL, 0.40 mmol, 3 eq.) and 3-thiophenesulfonyl
3
chloride (25 mg, 0.14 mmol, 1.05 eq.) in DCM at 25°C for 16 h. The
1
2.05 (dt, J=15.2, 7.5 Hz, 1H), 1.99–1.80 (m, 2H), 1.74 (ddd, J=14.9,
9.7, 5.0 Hz, 3H), 1.61–1.48 (m, 2H). C NMR (CDCl
product was obtained as a pale brown oil (48 mg, 69%). H NMR
13
): δ 151.6, 148.7,
(CDCl ): δ 7.83 (dd, J=3.1, 1.3 Hz, 1H), 7.22 (dd, J=5.1, 3.1 Hz, 1H),
3
3
1
1
2
5
43.1, 139.9, 134.2, 133.0, 132.2, 129.4, 117.8, 117.4, 112.2, 102.9,
7.14 (dd, J=5.1, 1.3 Hz, 1H), 6.72 (d, J=8.2 Hz, 1H), 6.43 (d, J=
8.2 Hz, 1H), 6.18 (s, 1H), 5.87 (d, J=1.4 Hz, 1H), 5.85 (d, J=1.4 Hz,
1H), 5.63–5.57 (m, 1H), 5.56 (d, J=4.3 Hz, 1H), 5.03 (d, J=12.4 Hz,
1H), 4.90 (dd, J=12.4, 2.7 Hz, 1H), 3.82 (s, 3H), 3.82 (s, 3H), 3.79 (s,
00.9, 85.5, 71.8, 62.0, 60.1, 59.4, 56.4, 55.7, 40.3, 28.3, 28.1, 27.8,
+
5.9, 25.8. HR-ESMS calcd. for C H NO S [M+H] 518.1843, found
18.1835.
26
32
8
3
H), 3.41 (ddd, J=12.9, 6.1, 5.1 Hz, 1H), 3.01–2.90 (m, 1H), 2.53 (dt,
(
R)-6-(Cyclohexylsulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihydroiso-
13
J=16.4, 5.1 Hz, 1H), 2.45–2.35 (m, 1H). C NMR (CDCl ): δ 151.5,
3
benzofuran-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]
isoquinoline (14k). Compound 14k was synthesised from 11
148.6, 143.1, 140.1, 140.0, 134.0, 132.8, 132.3, 130.1, 129.4, 127.4,
1
25.5, 117.8, 116.6, 112.1, 102.6, 100.8, 85.8, 71.8, 60.0, 59.3, 56.4,
(
67 mg, 0.17 mmol, 1 eq.) according to General Procedure A, with
+
55.8, 40.5, 26.8. HR-ESMS calcd. for C H NO S [M+H] 532.1094,
2
5
26
8 2
DBU (78 μL, 0.52 mmol, 3 eq.) and cyclohexanesulfonyl chloride
found 532.1086.
(52 μL, 0.36 mmol, 2.05 eq.) in DCM at 25°C for 120 h. The product
1
was obtained as a colourless oil (13 mg, 14%). H NMR (CDCl ): δ
(R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-6-(furan-
2-ylsulfonyl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]iso-
quinoline (14o). Compound 14o was synthesised from 11 (76 mg,
3
6
1
4
3
.74–6.62 (m, 1H), 6.34 (s, 1H), 6.21 (d, J=8.2 Hz, 1H), 5.91 (d, J=
.4 Hz, 1H), 5.90 (d, J=1.4 Hz, 1H), 5.69–5.61 (m, 1H), 5.40 (d, J=
.0 Hz, 1H), 5.09 (d, J=1.9 Hz, 2H), 3.86 (s, 3H), 3.82 (s, 3H), 3.82 (s,
H), 3.57–3.42 (m, 1H), 3.15–3.01 (m, 1H), 2.86–2.73 (m, 2H), 2.61–
0.20 mmol, 1 eq.) according to General Procedure A, with Et N
3
(83 μL, 0.59 mmol, 3 eq.) and 2-furansulfonyl chloride (23 μL,
0.21 mmol, 1.05 eq.) in DCM at 25°C for 16 h. The product was
2.46 (m, 1H), 2.24–2.13 (m, 1H), 1.93–1.76 (m, 3H), 1.72–1.59 (m, 2H),
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© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
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1
1
obtained as a white foam (45 mg, 44%). H NMR (CDCl ): δ 7.33 (s,
obtained as a white foam (100 mg, 78%). H NMR (CDCl ): δ 7.73
3
3
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
1
H), 6.92 (d, J=3.4 Hz, 1H), 6.70 (d, J=8.2 Hz, 1H), 6.40–6.31 (m,
H), 6.22 (s, 1H), 5.88 (d, J=1.0 Hz, 1H), 5.85 (d, J=1.1 Hz, 1H), 5.63–
(dd, J=5.2, 3.3 Hz, 2H), 7.43 (ddd, J=6.7, 3.9, 1.2 Hz, 1H), 7.39–7.31
(m, 2H), 6.68 (d, J=8.2 Hz, 1H), 6.33 (d, J=8.2 Hz, 1H), 6.12 (s, 1H),
5.85 (d, J=1.4 Hz, 1H), 5.83 (d, J=1.4 Hz, 1H), 5.58 (s, 2H), 4.98 (d,
J=12.4 Hz, 1H), 4.82 (d, J=13.8 Hz, 1H), 3.83 (s, 3H), 3.80 (s, 3H),
3.76 (s, 3H), 3.40 (ddd, J=13.5, 6.5, 4.2 Hz, 1H), 2.83 (ddd, J=13.7,
2
5.59 (m, 1H), 5.48 (d, J=4.1 Hz, 1H), 5.02 (dt, J=12.4, 7.5 Hz, 2H),
3.82 (s, 3H), 3.81 (s, 3H), 3.80 (s, 3H), 3.55 (dt, J=13.5, 5.5 Hz, 1H),
13
2
.95–2.81 (m, 1H), 2.57–2.42 (m, 2H). C NMR (CDCl ): δ 151.6,
3
1
3
148.7, 148.6, 145.7, 143.2, 140.0, 134.1, 132.7, 132.3, 129.2, 117.7,
9.5, 5.6 Hz, 1H), 2.39 (ddd, J=16.3, 12.8, 9.2 Hz, 2H). C NMR
1
5
16.4, 116.2, 112.1, 111.2, 102.6, 100.8, 85.4, 72.0, 60.1, 59.3, 56.4,
6.0, 40.6, 27.0. HR-ESMS calcd. for C H NO S [M+H] 516.1323,
(CDCl ): δ 151.4, 148.5, 143.0, 140.4, 139.9, 133.9, 132.7, 132.3, 132.2,
3
+
129.1, 128.7 (2×), 127.1 (2×), 117.6, 116.4, 112.1, 102.5, 100.7, 85.5,
25
26
9
found 516.1329.
71.6, 59.9, 59.2, 56.3, 55.6, 40.1, 26.7. HR-ESMS calcd. for
+
C
H
28NO
S
[M+H] 526.1530, found 526.1527.
27
8
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-6-(furan-
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
3
-ylsulfonyl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]iso-
(R)-6-(Benzylsulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihydroisobenzo-
furan-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]isoqui-
noline (14t). Compound 14t was synthesised from 11 (68 mg,
quinoline (14p). Compound 14p was synthesised from 11 (95 mg,
0.25 mmol, 1 eq.) according to General Procedure A, with Et N
(103 μL, 0.74 mmol, 3 eq.) and 3-furansulfonyl chloride (28 μL,
3
0.18 mmol, 1 eq.) according to General Procedure A, with Et N
3
0
.26 mmol, 1.05 eq.) in DCM at 25°C for 16 h. The product was
(74 μL, 0.53 mmol, 3 eq.) and phenylmethanesulfonyl chloride
1
obtained as a yellow oil (42 mg, 33%). H NMR (CDCl ): δ 7.80 (d,
J=0.5 Hz, 1H), 7.31 (t, J=1.7 Hz, 1H), 6.74 (d, J=8.2 Hz, 1H), 6.50 (d,
J=8.1 Hz, 1H), 6.41 (d, J=1.1 Hz, 1H), 6.23 (s, 1H), 5.87 (d, J=1.2 Hz,
(50 mg, 0.26 mmol, 1.5 eq.) in DCM at 50°C for 6 h. The product
3
1
was obtained as a colourless oil (25 mg, 26%). H NMR (CDCl ): δ
3
7.31–7.22 (m, 5H), 6.74 (d, J=8.2 Hz, 1H), 6.38 (d, J=8.2 Hz, 1H),
6.33 (s, 1H), 5.94 (d, J=1.4 Hz, 1H), 5.91 (d, J=1.4 Hz, 1H), 5.61 (dd,
J=2.5, 1.4 Hz, 1H), 5.32 (d, J=3.9 Hz, 1H), 5.16–5.02 (m, 2H), 4.31 (d,
J=13.8 Hz, 1H), 4.16 (d, J=13.8 Hz, 1H), 3.86 (s, 3H), 3.85 (s, 3H),
3.79 (s, 3H), 3.52–3.33 (m, 1H), 2.93 (ddd, J=13.7, 8.4, 6.8 Hz, 1H),
1
H), 5.85 (d, J=1.3 Hz, 1H), 5.61–5.55 (m, 1H), 5.50 (d, J=4.2 Hz,
1H), 5.04 (d, J=12.3 Hz, 1H), 4.92 (dd, J=12.4, 2.5 Hz, 1H), 3.82 (s,
3
H), 3.79 (s, 3H), 3.79 (s, 3H), 3.39–3.27 (m, 1H), 3.11–2.99 (m, 1H),
13
2.65 (dt, J=16.0, 5.5 Hz, 1H), 2.59–2.47 (m, 1H). C NMR (CDCl ): δ
3
1
3
1
1
5
51.5, 148.7, 145.4, 144.4, 143.0, 139.9, 134.0, 132.7, 132.2, 129.6,
2.61 (dd, J=9.0, 6.3 Hz, 2H). C NMR (CDCl ): δ 151.5, 148.7, 143.1,
3
26.7, 117.8, 116.6, 112.1, 108.3, 102.5, 100.8, 86.0, 71.8, 60.0, 59.2,
139.9, 133.9, 132.9, 132.1, 130.8 (2×), 129.4, 129.1, 128.5 (2×), 128.4,
117.8, 116.9, 112.2, 102.7, 100.8, 85.8, 71.7, 60.1, 59.1, 58.3, 56.4,
+
6.3, 55.7, 40.9, 27.0. HR-ESMS calcd. for C H NO S [M+H]
25
26
9
+
516.1323, found 516.1327.
55.6, 40.6, 27.6. HR-ESMS calcd. for C H NO S [M+H] 540.1687,
2
8
30
8
found 540.1692.
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-((1-methyl-1H-imidazol-4-yl)sulfonyl)-5,6,7,8-tetrahydro
1,3]dioxolo[4,5-g]isoquinoline (14q). Compound 14q was syn-
thesised from 11 (79 mg, 0.21 mmol, 1 eq.) according to General
(R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-tosyl-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]isoquinoline
(14u). Compound 14u was synthesised from 11 (76 mg, 0.20 mmol,
[
Procedure A, with Et N (86 μL, 0.62 mmol, 3 eq.) and 1-methyl-1H-
imidazole-4-sulfonyl chloride (24 mg, 0.22 mmol, 1.05 eq.) in DCM
1 eq.) according to General Procedure A, with Et N (82 μL,
3
3
0.59 mmol, 3 eq.) and 4-toluenesulfonyl chloride (56 mg,
at 25°C for 16 h. The product was obtained as a white foam
0.30 mmol, 1.5 eq.) in DCM at 50°C for 6 h. The product was
1
1
(
49 mg, 45%). H NMR (CDCl ): δ 7.43 (s, 1H), 7.41 (s, 1H), 6.68 (d,
obtained as a colourless oil (69 mg, 65%). H NMR (CDCl ): δ 7.62 (d,
3
3
J=8.2 Hz, 1H), 6.28–6.17 (m, 2H), 5.92–5.87 (m, 2H), 5.62 (s, 1H),
5.56 (s, 1H), 5.03 (q, J=12.4 Hz, 2H), 3.84 (s, 3H), 3.80 (s, 6H), 3.66 (s,
3H), 3.44–3.34 (m, 1H), 2.89–2.74 (m, 1H), 2.58–2.46 (m, 1H), 2.46–
J=8.3 Hz, 2H), 7.15 (d, J=8.1 Hz, 2H), 6.70 (d, J=8.2 Hz, 1H), 6.39
(d, J=8.2 Hz, 1H), 6.15 (s, 1H), 5.87 (d, J=1.4 Hz, 1H), 5.85 (d, J=
1.4 Hz, 1H), 5.62–5.55 (m, 2H), 5.01 (d, J=12.4 Hz, 1H), 4.88 (dd, J=
12.4, 2.4 Hz, 1H), 3.83 (s, 3H), 3.82 (s, 3H), 3.79 (s, 3H), 3.37 (ddd, J=
13.3, 6.4, 4.6 Hz, 1H), 2.92–2.82 (m, 1H), 2.46 (dt, J=16.3, 5.1 Hz, 1H),
13
2
1
1
.33 (m, 1H). C NMR (CDCl ): δ 151.6, 149.0, 143.2, 141.8, 139.7,
36.0, 134.1, 132.4, 132.2, 129.2, 129.1, 117.6, 116.1, 112.2, 102.8,
01.0, 85.4, 71.8, 60.1, 59.3, 56.4, 55.5, 40.0, 33.1, 27.0. HR-ESMS
3
1
3
2.39 (dd, J=9.2, 6.8 Hz, 1H), 2.34 (s, 3H). C NMR (CDCl ): δ 151.5,
3
+
calcd. for C H N O S [M+H] 530.1592, found 530.1600.
148.5, 143.1, 143.0 (2×), 140.0, 137.6, 134.0, 132.9, 132.3, 129.4 (2×),
25
28
3
8
1
27.3 (2×), 117.8, 116.7, 112.1, 102.6, 100.8, 85.8, 71.8, 60.0, 59.3,
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-((1-methyl-1H-pyrazol-4-yl)sulfonyl)-5,6,7,8-tetrahydro
1,3]dioxolo[4,5-g]isoquinoline (14r). Compound 14r was synthes-
+
56.4, 55.7, 40.3, 26.9, 21.6. HR-ESMS calcd. for C H NO S [M+H]
2
8
30
8
540.1687, found 540.1688.
[
ised from 11 (86 mg, 0.22 mmol, 1 eq.) according to General
Procedure A, with Et N (93 μL, 0.66 mmol, 3 eq.) and 1-methyl-1H-
pyrazole-4-sulfonyl chloride (26 μL, 0.23 mmol, 1.05 eq.) in DCM at
(R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-((2-methoxyphenyl)sulfonyl)-5,6,7,8-tetrahydro[1,3]diox-
olo[4,5-g]isoquinoline (14v). Compound 14v was synthesised from
11 (97 mg, 0.25 mmol, 1 eq.) according to General Procedure A,
3
2
4
1
5
4
5°C for 16 h. The product was obtained as a white foam (49 mg,
1
2%). H NMR (CDCl ): δ 7.62 (s, 1H), 7.57 (s, 1H), 6.76 (d, J=8.2 Hz,
with Et N (105 μL, 0.75 mmol, 3 eq.) and 2-methoxybenzenesulfonyl
3
3
H), 6.60 (d, J=8.1 Hz, 1H), 6.22 (s, 1H), 5.85 (d, J=10.9 Hz, 2H),
.60–5.53 (m, 1H), 5.46 (d, J=4.2 Hz, 1H), 5.01 (d, J=12.3 Hz, 1H),
.85 (dd, J=12.3, 2.5 Hz, 1H), 3.83 (s, 3H), 3.81 (s, 3H), 3.79 (s, 3H),
chloride (78 mg, 0.38 mmol, 1.5 eq.) in DCM at 50°C for 6 h. The
1
product was obtained as a colourless oil (68 mg, 49%). H NMR
(CDCl ): δ 7.93 (dd, J=7.8, 1.7 Hz, 1H), 7.41 (td, J=8.4, 1.7 Hz, 1H),
3
3.76 (s, 3H), 3.32–3.21 (m, 1H), 3.19–3.09 (m, 1H), 2.71 (dt, J=15.8,
6.97 (t, J=7.3 Hz, 1H), 6.78 (d, J=8.0 Hz, 1H), 6.64 (d, J=8.2 Hz, 1H),
6.24–6.14 (m, 2H), 5.90 (d, J=1.4 Hz, 1H), 5.88 (d, J=1.4 Hz, 1H),
5.75–5.66 (m, 2H), 5.19 (dd, J=12.4, 2.7 Hz, 1H), 5.09 (d, J=12.4 Hz,
1H), 3.91 (s, 3H), 3.83 (s, 3H), 3.81 (s, 3H), 3.44 (s, 3H), 3.36 (ddd, J=
13
5.9 Hz, 1H), 2.57–2.45 (m, 1H). C NMR (CDCl
): δ 151.5, 148.6, 143.0,
40.0, 138.4, 134.0, 132.8, 132.2, 131.7, 129.9, 121.7, 118.0, 116.7,
12.2, 102.5, 100.8, 86.4, 71.7, 60.1, 59.2, 56.4, 55.7, 41.2, 39.5, 27.1.
3
1
1
+
HR-ESMS calcd. for C H N O S [M+H] 530.1592, found 530.1597.
13.9, 6.6, 2.8 Hz, 1H), 2.64–2.54 (m, 1H), 2.29–2.19 (m, 1H), 2.16–2.04
25
28
3
8
13
(
m, 1H). C NMR (CDCl ): δ 156.8, 151.4, 148.2, 143.3, 139.8, 134.2,
3
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
1
33.9, 133.2, 132.5, 131.7, 129.3, 128.6, 120.1, 117.6, 117.5, 112.0,
thoxy-6-(phenylsulfonyl)-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]iso-
quinoline (14s). Compound 14s was synthesised from 11 (94 mg,
111.6, 102.5, 100.8, 85.4, 72.0, 60.1, 59.2, 56.4, 56.3, 55.2, 39.5, 26.7.
HR-ESMS calcd. for C H NO S [M+H] 556.1636, found 556.1643.
+
28 30
9
0
.25 mmol, 1 eq.) according to General Procedure A, with Et N
3
(102 μL, 0.74 mmol, 3 eq.) and benzenesulfonyl chloride (47 μL,
(R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-((3-methoxyphenyl)sulfonyl)-5,6,7,8-tetrahydro[1,3]diox-
0.37 mmol, 1.5 eq.) in DCM at 50°C for 6 h. The product was
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10
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olo[4,5-g]isoquinoline (14w). Compound 14 w was synthesised
from 11 (87 mg, 0.23 mmol, 1 eq.) according to General Procedure
71.7, 60.1, +59.3, 56.4, 55.8, 40.4, 26.7. HR-ESMS calcd. for
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
C H ClNO S [M+H] 560.1140, found 560.1141.
2
7
27
8
A, with Et N (94 μL, 0.68 mmol, 3 eq.) and 3-methoxybenzenesul-
3
(
R)-6-((4-Chlorophenyl)sulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihy-
droisobenzofuran-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo
4,5-g]isoquinoline (14aa). Compound 14aa was synthesised from
1 (68 mg, 0.18 mmol, 1 eq.) according to General Procedure A,
with Et N (73 μL, 0.53 mmol, 3 eq.) and 4-chlorobenzenesulfonyl
fonyl chloride (48 μL, 0.34 mmol, 1.5 eq.) in DCM at 50°C for 16 h.
1
The product was obtained as a colourless oil (56 mg, 45%). H NMR
[
1
(
CDCl ): δ 7.35–7.31 (m, 1H), 7.25 (t, J=8.0 Hz, 1H), 7.23–7.20 (m,
3
1
H), 6.97 (ddd, J=8.2, 2.6, 1.0 Hz, 1H), 6.72 (d, J=8.2 Hz, 1H), 6.43
d, J=8.2 Hz, 1H), 6.15 (s, 1H), 5.86 (d, J=1.4 Hz, 1H), 5.85 (d, J=
.4 Hz, 1H), 5.63–5.55 (m, 2H), 5.02 (d, J=12.4 Hz, 1H), 4.88 (dd, J=
3
(
1
chloride (56 mg, 0.26 mmol, 1.5 eq.) in DCM at 50
product was obtained as a white foam (72 mg, 73%). H NMR
°
C for 6 h. The
1
12.4, 2.1 Hz, 1H), 3.82 (d, J=1.6 Hz, 6H), 3.79 (s, 3H), 3.73 (s, 3H),
3.41 (ddd, J=13.4, 6.6, 4.5 Hz, 1H), 2.99–2.88 (m, 1H), 2.49 (dt, J=
(
CDCl ): δ 7.70–7.66 (m, 2H), 7.36–7.31 (m, 2H), 6.71 (d, J=8.2 Hz,
3
1
3
1H), 6.36 (d, J=8.2 Hz, 1H), 6.17 (s, 1H), 5.88 (d, J=1.4 Hz, 1H), 5.87
1
6.4, 5.1 Hz, 1H), 2.38 (ddd, J=16.2, 9.1, 6.9 Hz, 1H). C NMR
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
(
(
d, J=1.4 Hz, 1H), 5.63–5.53 (m, 2H), 5.01 (d, J=12.4 Hz, 1H), 4.84
dd, J=12.4, 2.7 Hz, 1H), 3.85 (s, 3H), 3.82 (s, 3H), 3.79 (s, 3H), 3.44–
(
CDCl ): δ 159.7, 151.6, 148.6, 143.1, 141.6, 140.0, 134.0, 132.9, 132.3,
29.8, 129.4, 119.5, 119.2, 117.8, 116.7, 112.2, 111.6, 102.6, 100.8,
3
1
8
3.34 (m, 1H), 2.89 (ddd, J=13.6, 9.3, 5.6 Hz, 1H), 2.50 (dt, J=16.5,
5.0 Hz, 1H), 2.40 (ddd, J=16.2, 9.2, 6.7 Hz, 1H). C NMR (CDCl
5.8, 71.8, 60.1, 59.3, 56.4, 55.9, 55.6, 40.4, 26.7. HR-ESMS calcd. for
1
3
+
3
): δ
C H NO S [M+H] 556.1636, found 556.1640.
2
8
30
9
1
51.6, 148.7, 143.1, 134.0, 139.1, 138.8, 134.0, 132.7, 132.2, 129.1,
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
129.1 (2×), 128.7 (2×), 117.7, 116.4, 112.2, 102.7, 100.9, 85.6, 71.7,
60.1, 59.3, 56.4, 55.8, 40.4, 26.9. HR-ESMS calcd. for C H ClNO S
+
thoxy-6-((4-methoxyphenyl)sulfonyl)-5,6,7,8-tetrahydro[1,3]diox-
2
7
27
8
olo[4,5-g]isoquinoline (14x). Compound 14x was synthesised from
[M+H] 560.1140, found 560.1138.
1
1 (116 mg, 0.30 mmol, 1 eq.) according to General Procedure A,
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
with Et N (126 μL, 0.90 mmol, 3 eq.) and 4-methoxybenzenesulfonyl
3
thoxy-7,8-dihydro[1,3]dioxolo[4,5-g]isoquinoline-6(5H)-sulfona-
mide (15a). To a chilled (À 5°C), stirring solution of chlorosulfony-
lisocyanate (43 μL, 0.49 mmol, 1.2 eq.) in anhydrous DCM (2 mL)
was added benzyl alcohol (51 μL, 0.49 mmol, 1.2 eq.) in a dropwise
chloride (93 mg, 0.45 mmol, 1.5 eq.) in DCM at 50°C for 6 h. The
1
product was obtained as a white foam (95 mg, 57%). H NMR
(
1
CDCl ): δ 7.70–7.63 (m, 2H), 6.85–6.79 (m, 2H), 6.71 (d, J=8.2 Hz,
H), 6.40 (d, J=8.2 Hz, 1H), 6.15 (s, 1H), 5.87 (d, J=1.4 Hz, 1H), 5.85
3
manner. The reaction was left to stir at À 5
°
C for 1 h before addition
(
d, J=1.4 Hz, 1H), 5.61–5.54 (m, 2H), 5.02 (d, J=12.3 Hz, 1H), 4.91
of mixture 11 (159 mg, 0.41 mmol, 1 eq.) and Et N (115 μL,
3
(dd, J=12.4, 2.6 Hz, 1H), 3.83 (s, 3H), 3.82 (s, 3H), 3.80 (s, 3H), 3.79 (s,
3H), 3.37 (ddd, J=13.1, 6.2, 4.7 Hz, 1H), 2.94–2.83 (m, 1H), 2.47 (dt,
0.82 mmol, 2 eq.) in anhydrous DCM (1 mL). The reaction mixture
1
3
was then left to stir at 25°C for 16 h. The reaction was washed with
J=16.3, 5.0 Hz, 1H), 2.37 (ddd, J=16.2, 9.2, 6.7 Hz, 1H). C NMR
CDCl ): δ 162.7, 151.5, 148.5, 143.1, 140.1, 134.0, 133.0, 132.3, 132.3,
0
.1 M HCl (3×10 mL), dried over MgSO , filtered and the filtrate
4
(
3
evaporated under reduced pressure, yielding the crude as a
colourless oil. In a three-neck flask, Pd/C (10%) was dispersed in
1
7
29.4 (2×), 129.3, 117.8, 116., 113.9 (2×), 112.1, 102.6, 100.8, 85.8,
1.8, 60.1, 59.3, 56.4, 55.7, 55.7, 40.2, 26.8. HR-ESMS calcd. for
+
MeOH (5 mL) and AcOH (1 mL) under N . The mixture was degassed
2
C H NO S [M+H] 556.1636, found 556.1634.
2
8
30
9
and saturated with H2 thrice. The crude was then added to the
reaction, and left to stir at 25°C under hydrogen atmosphere
(balloon) for 16 h. The catalyst was removed by gravity filtration,
and the filtrate was evaporated under reduced pressure. Purifica-
(
R)-6-((2-Chlorophenyl)sulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihy-
droisobenzofuran-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo
4,5-g]isoquinoline (14y). Compound 14y was synthesised from 11
(74 mg, 0.19 mmol, 1 eq.) according to General Procedure A, with
Et N (80 μL, 0.57 mmol, 3 eq.) and 2-chlorobenzenesulfonyl chloride
[
tion via gradient column chromatography (DCM: 1 to 5% MeOH)
1
afforded 15a as a white solid (22 mg, 58%). H NMR (CDCl
): δ 6.66
3
3
(
39 μL, 0.29 mmol, 1.5 eq.) in DCM at 50°C for 6 h. The product was
(d, J=8.2 Hz, 1H), 6.34 (s, 1H), 6.10 (d, J=8.1 Hz, 1H), 5.92 (d, J=
1.4 Hz, 1H), 5.91 (d, J=1.4 Hz, 1H), 5.74–5.68 (m, 1H), 5.46 (d, J=
4.5 Hz, 1H), 5.25–5.05 (m, 2H), 3.93 (s, 3H), 3.83 (s, 3H), 3.82 (s, 3H),
1
obtained as a colourless oil (64 mg, 60%). H NMR (CDCl ): δ 7.73 (t,
J=1.7 Hz, 1H), 7.63 (d, J=7.9 Hz, 1H), 7.44–7.39 (m, 1H), 7.30 (t, J=
3
7
1
.9 Hz, 1H), 6.72 (d, J=8.2 Hz, 1H), 6.40 (d, J=8.2 Hz, 1H), 6.14 (s,
H), 5.92–5.82 (m, 2H), 5.63–5.52 (m, 2H), 5.00 (d, J=12.4 Hz, 1H),
3.62–3.50 (m, 1H), 2.88–2.76 (m, 1H), 2.69–2.59 (m, 1H), 2.53 – 2.41
13
(m, 1H). C NMR (CDCl ): δ 151.6, 148.8, 143.2, 140.0, 134.2, 132.5,
3
4.81 (dd, J=12.4, 2.6 Hz, 1H), 3.87 (s, 3H), 3.83 (s, 3H), 3.79 (s, 3H),
3.44 (ddd, J=13.3, 6.7, 4.4 Hz, 1H), 3.01–2.90 (m, 1H), 2.51 (dt, J=
132.1, 129.6, 117.7, 116.3, 112.3, 102.9, 101.0, 84.9, 71.8, 60.2, 59.5,
+
56.4, 55.8, 40.4, 27.6. HR-ESMS calcd. for C21
465.1326, found 465.1333.
H
N
2
O S
8
[M+H]
25
1
3
1
6.4, 5.2 Hz, 1H), 2.43 (ddd, J=16.4, 9.0, 7.1 Hz, 1H). C NMR
(
CDCl ): δ 151.5, 148.6, 143.3, 139.9, 138.5, 134.2, 133.3, 132.7, 132.4,
32.2, 132.2, 131.8, 129.1, 126.9, 117.6, 116.9, 112.1, 102.8, 100.9,
3
(R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-N-methyl-7,8-dihydro[1,3]dioxolo[4,5-g]isoquinoline-6(5H)-
sulfonamide (15b). Compound 15b was synthesised from 11
1
8
5.0, 71.8, 60.1, 59.4, 56.4, 56.0, 39.9, 27.5. HR-ESMS calcd. for
+
C H ClNO S [M+H] 560.1140, found 560.1138.
2
7
27
8
(
103 mg, 0.27 mmol, 1 eq.) according to General Procedure A, with
3
(
R)-6-((3-Chlorophenyl)sulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihy-
droisobenzofuran-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo
4,5-g]isoquinoline (14z). Compound 14z was synthesised from 11
(80 mg, 0.21 mmol, 1 eq.) according to General Procedure A, with
Et N (87 μL, 0.62 mmol, 3 eq.) and 3-chlorobenzenesulfonyl chloride
Et N (111 μL, 0.80 mmol, 3 eq.) and methylsulfamoyl chloride
(35 μL, 0.40 mmol, 1.5 eq.) in DCM at 25°C for 16 h. The product
1
[
was obtained as a colourless oil (63 mg, 50%). H NMR (CDCl ): δ
3
6.68 (d, J=8.2 Hz, 1H), 6.33 (s, 1H), 6.19 (d, J=8.3 Hz, 1H), 5.89 (s,
2H), 5.70–5.62 (m, 1H), 5.39 (d, J=4.3 Hz, 1H), 5.19–5.05 (m, 2H),
3
(
44 μL, 0.31 mmol, 1.5 eq.) in DCM at 50°C for 6 h. The product was
3.87 (s, 3H), 3.81 (s, 6H), 3.52–3.44 (m, 1H), 2.87–2.69 (m, 2H), 2.62 (s,
1
13
obtained as a colourless oil (57 mg, 49%). H NMR (CDCl ): δ 8.11 (d,
J=7.5 Hz, 1H), 7.46–7.40 (m, 2H), 7.39–7.33 (m, 1H), 6.63 (d, J=
3H), 2.51 (dt, J=7.0, 3.8 Hz, 1H). C NMR (CDCl ): δ 151.5, 148.7,
3
3
143.1, 140.0, 134.1, 132.8, 132.2, 129.5, 117.7, 116.9, 112.2, 102.8,
100.9, 85.2, 71.8, 60.1, 59.3, 56.3, 56.0, 40.4, 29.4, 27.4. HR-ESMS
8
5
.2 Hz, 1H), 6.25 (s, 1H), 6.08 (d, J=8.2 Hz, 1H), 5.95–5.88 (m, 2H),
.68–5.62 (m, 2H), 5.08 (dd, J=12.4, 2.5 Hz, 1H), 4.99 (d, J=12.4 Hz,
+
calcd. for C H N O S [M+H] 479.1483, found 479.1487.
2
2
27
2
8
1H), 3.89 (s, 3H), 3.83 (s, 3H), 3.81 (s, 3H), 3.50 (ddd, J=13.6, 6.2,
1
3
(R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-N-ethyl-
-methoxy-7,8-dihydro[1,3]dioxolo[4,5-g]isoquinoline-6(5H)-sul-
2
1
1
.8 Hz, 1H), 2.75–2.64 (m, 1H), 2.48–2.32 (m, 2H). C NMR (CDCl ): δ
3
4
51.6, 148.7, 143.1, 142.1, 140.0, 134.9, 134.0, 132.7, 132.4, 132.2,
30.0, 129.0, 127.5, 125.3, 117.8, 116.4, 112.2, 102.6, 100.9, 85.6,
fonamide (15c). Compound 15cc was synthesised from 11
(113 mg, 0.27 mmol, 1 eq.) according to General Procedure A, with
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11
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Et N (122 μL, 0.88 mmol, 3 eq.) and ethylsulfamoyl chloride (45 μL,
2H), 5.67–5.60 (m, 1H), 5.39 (d, J=4.3 Hz, 1H), 5.15–5.04 (m, 2H),
3.85 (s, 3H), 3.82 (s, 3H), 3.81 (s, 3H), 3.55–3.36 (m, 1H), 2.90–2.66 (m,
4H), 2.60–2.46 (m, 1H), 1.70 (dd, J=13.4, 6.7 Hz, 1H), 0.87 (d, J=
3
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
0
.44 mmol, 1.5 eq.) in DMF at 25°C for 16 h. The product was
1
obtained as a yellow oil (17 mg, 13%). H NMR (CDCl ): δ 6.68 (d,
J=8.2 Hz, 1H), 6.33 (s, 1H), 6.21 (d, J=8.1 Hz, 1H), 5.90 (s, 2H), 5.69–
5.62 (m, 1H), 5.40 (d, J=4.2 Hz, 1H), 5.17–5.04 (m, 2H), 3.87 (s, 3H),
3
1
3
6.7 Hz, 6H). C NMR (CDCl ): δ 151.5, 148.7, 143.1, 140.0, 134.1,
3
132.9, 132.2, 129.6, 117.8, 117.0, 112.2, 102.8, 100.8, 85.3, 71.8, 60.1,
59.3, 56.4, 55.9, 50.7, 40.5, 28.5, 27.4, 20.1, 20.1. HR-ESMS calcd. for
3
2
.82 (s, 6H), 3.54–3.45 (m, 1H), 3.11–2.89 (m, 2H), 2.87–2.68 (m, 2H),
.52 (dt, J=6.6, 3.6 Hz, 1H), 1.12 (t, J=7.2 Hz, 3H). C NMR (CDCl3):
13
+
C H N O S [M+H] 521.1952, found 521.1946.
25
33
2
8
δ 151.6, 148.7, 143.2, 140.0, 134.1, 132.9, 132.2, 129.6, 117.7, 116.9,
12.3, 102.9, 100.9, 85.3, 71.8, 60.2, 59.4, 56.4, 55.9, 40.5, 38.3, 27.5,
(R)-N-cyclopropyl-5-((S)-4,5-dimethoxy-1,3-dihydroisobenzofuran-
1-yl)-4-methoxy-7,8-dihydro[1,3]dioxolo[4,5-g]isoquinoline-6(5H)-
sulfonamide (15h). Compound 15h was synthesised from 11
1
+
15.3. HR-ESMS calcd. for C H N O S [M+H] 493.1639, found
23
29
2
8
493.1643.
(
78 mg, 0.20 mmol, 1 eq.) according to General Procedure A, with
3
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-N,N-di-
Et N (85 μL, 0.61 mmol, 3 eq.) and cyclopropylsulfamoyl chloride
ethyl-4-methoxy-7,8-dihydro[1,3]dioxolo[4,5-g]isoquinoline-
6(5H)-sulfonamide (15d). Compound 15d was synthesised from 11
(102 mg, 0.26 mmol, 1 eq.) according to General Procedure A, with
DBU (79 μL, 0.79 mmol, 3 eq.) and diethylsulfamoyl chloride (53 μL,
(21 μL, 0.21 mmol, 1.05 eq.) in DCM at 25°C for 16 h. The product
1
was obtained as a colourless oil (44 mg, 43%). H NMR (CDCl ): δ
3
6.72 (d, J=8.2 Hz, 1H), 6.40 (d, J=8.2 Hz, 1H), 6.34 (s, 1H), 5.89 (d,
J=1.4 Hz, 1H), 5.88 (d, J=1.4 Hz, 1H), 5.66 (dd, J=3.7, 2.8 Hz, 1H),
5.46 (d, J=3.9 Hz, 1H), 5.06 (d, J=12.3 Hz, 1H), 4.99 (dd, J=12.3,
2.6 Hz, 1H), 4.86 (br s, 1H), 3.83 (s, 3H), 3.80 (s, 3H), 3.77 (s, 3H),
3.50–3.42 (m, 1H), 3.04–2.93 (m, 1H), 2.83–2.74 (m, 1H), 2.70 (dt, J=
16.1, 5.3 Hz, 1H), 2.41–2.32 (m, 1H), 0.70–0.61 (m, 1H), 0.60–0.49 (m,
0
.40 mmol, 1.5 eq.) in DMF at 60°C for 16 h. The product was
1
obtained as a colourless oil (17 mg, 10%). H NMR (CDCl ): δ 6.71 (d,
J=8.2 Hz, 1H), 6.35 (d, J=8.2 Hz, 1H), 6.33 (s, 1H), 5.88 (d, J=1.4 Hz,
3
1
H), 5.87 (d, J=1.4 Hz, 1H), 5.65–5.60 (m, 1H), 5.34 (d, J=3.9 Hz,
1
3
1H), 5.10–5.01 (m, 2H), 3.83 (s, 3H), 3.81 (s, 3H), 3.79 (s, 3H), 3.41–
3H). C NMR (CDCl ): δ 151.5, 148.7, 143.1, 140.1, 134.1, 132.9,
3
3
1
1
7
.34 (m, 1H), 3.27–3.12 (m, 4H), 2.89–2.75 (m, 2H), 2.64–2.54 (m, 1H),
132.1, 130.1, 117.9, 117.1, 112.3, 102.7, 100.9, 86.2, 71.7, 60.1, 59.3,
13
+
.12 (t, J=7.1 Hz, 6H). C NMR (CDCl ): δ 151.6, 148.8, 143.2, 139.9,
56.4, 56.1, 41.1, 27.9, 24.4, 6.6, 6.4. HR-ESMS calcd. for C H N O S
3
24 29
2
8
34.1, 132.9, 132.2, 129.3, 117.6, 116.8, 112.3, 102.8, 100.9, 85.4,
[M+H] 505.1639, found 505.1641.
1.9, 66.5 (2×), 60.1, 59.3, 56.4, 56.3, 46.4, 40.6 (2×), 27.3. HR-ESMS
+
(R)-N-cyclopentyl-5-((S)-4,5-dimethoxy-1,3-dihydroisobenzofuran-
calcd. for C H N O S [M+H] 521.1952, found 521.1956.
25
33
2
8
1
-yl)-4-methoxy-7,8-dihydro[1,3]dioxolo[4,5-g]isoquinoline-6(5H)-
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
sulfonamide (15i). Compound 15c was synthesised from 11
(113 mg, 0.29 mmol, 1 eq.) according to General Procedure A, with
DBU (87 μL, 0.88 mmol, 3 eq.) and cyclopentylsulfamoyl chloride
thoxy-N-propyl-7,8-dihydro[1,3]dioxolo[4,5-g]isoquinoline-6(5H)-
sulfonamide (15e). Compound 15e was synthesised from 11
(100 mg, 0.26 mmol, 1 eq.) according to General Procedure A, with
(57 μL, 0.44 mmol, 1.5 eq.) in DCM at 50 C for 16 h. The product
°
1
Et N (109 μL, 0.78 mmol, 3 eq.) and 1-propylsulfamoyl chloride
was obtained as a colourless oil (56 mg, 36%). H NMR (CDCl
): δ
3
3
(
47 μL, 0.39 mmol, 1.5 eq.) in DCM at 25°C for 16 h. The product
6.69 (d, J=8.2 Hz, 1H), 6.33 (s, 1H), 6.29 (d, J=8.2 Hz, 1H), 5.88 (s,
2H), 5.64 (d, J=3.4 Hz, 1H), 5.41 (d, J=4.1 Hz, 1H), 5.06 (s, 2H), 3.82
1
was obtained as a light yellow oil (81 mg, 61%). H NMR (CDCl ): δ
3
6
2
3
.68 (d, J=8.2 Hz, 1H), 6.32 (s, 1H), 6.23 (d, J=8.1 Hz, 1H), 5.88 (s,
H), 5.70–5.60 (m, 1H), 5.39 (d, J=4.2 Hz, 1H), 5.15–5.04 (m, 2H),
.86–3.83 (m, 3H), 3.81 (s, 3H), 3.81 (s, 3H), 3.54–3.43 (m, 1H), 3.02–
(s, 6H), 3.80 (s, 3H), 3.65–3.45 (m, 2H), 2.88–2.75 (m, 2H), 2.62–2.51
13
(m, 1H), 1.93–1.76 (m, 2H), 1.67–1.37 (m, 6H). C NMR (CDCl ): δ
3
151.5, 148.6, 143.0, 140.0, 134.0, 133.0, 132.1, 129.7, 117.7, 117.1,
2.71 (m, 4H), 2.58–2.44 (m, 1H), 1.55–1.42 (m, 2H), 0.87 (t, J=7.4 Hz,
112.2, 102.8, 100.8, 85.6, 71.7, 60.1, 59.3, 56.3, 55.9, 55.3, 40.4, 33.8,
1
3
+
3H). C NMR (CDCl
): δ 151.5, 148.6, 143.1, 140.0, 134.0, 132.9,
32.2, 129.6, 117.7, 117.0, 112.2, 102.8, 100.8, 85.3, 71.8, 60.1, 59.3,
6.3, 55.9, 45.0, 40.4, 27.4, 23.1, 11.3. HR-ESMS calcd. for
33.5, 27.5, 23.4, 23.3. HR-ESMS calcd. for C26
533.1952, found 533.1945.
H
N
2
O S
8
[M+H]
3
33
1
5
+
(R)-N-cyclohexyl-5-((S)-4,5-dimethoxy-1,3-dihydroisobenzofuran-
1-yl)-4-methoxy-7,8-dihydro[1,3]dioxolo[4,5-g]isoquinoline-6(5H)-
sulfonamide (15j). Compound 15j was synthesised from 11
(97 mg, 0.25 mmol, 1 eq.) according to General Procedure A, with
DBU (75 μL, 0.76 mmol, 3 eq.) and cyclohexylsulfamoyl chloride
C H N O S [M+H] 507.1796, found 507.1793.
2
4
31
2
8
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-N-
isopropyl-4-methoxy-7,8-dihydro[1,3]dioxolo[4,5-g]isoquinoline-
(5H)-sulfonamide (15f). Compound 15f was synthesised from 11
115 mg, 0.30 mmol, 1 eq.) according to General Procedure A, with
6
(
(58 μL, 0.38 mmol, 1.5 eq.) in DMF at 60°C for 16 h. The product
1
DBU (89 μL, 0.90 mmol, 3 eq.) and isopropylsulfamoyl chloride
was obtained as a colourless oil (16 mg, 12%). H NMR (CDCl ): δ
3
(54 μL, 0.45 mmol, 1.5 eq.) in DCM at 50°C for 16 h. The product
6.71 (d, J=8.2 Hz, 1H), 6.39–6.26 (m, 2H), 5.91–5.82 (m, 2H), 5.65–
5.58 (m, 1H), 5.41 (d, J=4.4 Hz, 1H), 5.08 (d, J=1.7 Hz, 2H), 3.84 (s,
3H), 3.83 (s, 3H), 3.82 (s, 3H), 3.54–3.45 (m, 1H), 3.17–3.04 (m, 1H),
2.92–2.77 (m, 2H), 2.64–2.51 (m, 1H), 1.68–1.48 (m, 5H), 1.28–1.07
1
was obtained as a yellow oil (56 mg, 37%). H NMR (CDCl
): δ 6.70
3
(
d, J=8.2 Hz, 1H), 6.33 (s, 1H), 6.29 (d, J=8.2 Hz, 1H), 5.89 (s, 2H),
.67–5.62 (m, 1H), 5.41 (d, J=4.2 Hz, 1H), 5.11–5.03 (m, 2H), 3.83 (s,
5
3
2
1
3
H), 3.82 (s, 3H), 3.81 (s, 3H), 3.53–3.43 (m, 2H), 2.90–2.75 (m, 2H),
(m, 5H). C NMR (CDCl ): δ 151.6, 148.7, 143.1, 140.1, 134.1, 133.1,
3
13
.62–2.51 (m, 1H), 1.16 (d, J=6.4 Hz, 3H), 1.11 (d, J=6.5 Hz, 3H).
C
132.2, 129.7, 117.8, 117.3, 112.3, 102.8, 100.8, 85.5, 71.7, 60.2, 59.3,
56.4, 55.9, 52.9, 40.6, 34.4, 34.2, 27.4, 25.5, 25.0, 25.0. HR-ESMS calcd.
NMR (CDCl ): δ 151.5, 148.6, 143.1, 140.0, 134.0, 133.0, 132.1, 129.7,
3
+
117.8, 117.1, 112.2, 102.8, 100.8, 85.5, 71.7, 60.1, 59.3, 56.4, 55.8,
for C H N O S [M+H] 547.2109, found 547.2114.
2
7
35
2
8
+
46.3, 40.4, 27.5, 24.1, 23.9. HR-ESMS calcd. for C24
07.1796, found 507.1799.
H
N
31
O S
8
[M+H]
2
(
R)-6-(Azetidin-1-ylsulfonyl)-5-((S)-4,5-dimethoxy-1,3-dihydroiso-
5
benzofuran-1-yl)-4-methoxy-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-g]
isoquinoline (15k). Compound 15k was synthesised from 11
(88 mg, 0.23 mmol, 1 eq.) according to General Procedure A, with
(R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-N-isobu-
tyl-4-methoxy-7,8-dihydro[1,3]dioxolo[4,5-g]isoquinoline-6(5H)-
sulfonamide (15g). Compound 15g was synthesised from 11
Et N (96 μL, 0.69 mmol, 3 eq.) and azetidine-1-sulfonyl chloride
3
(
99 mg, 0.26 mmol, 1 eq.) according to General Procedure A, with
(23 μL, 0.24 mmol, 1.05 eq.) in DCM at 25°C for 16 h. The product
1
DBU (76 μL, 0.77 mmol, 3 eq.) and isobutylsulfamoyl chloride
was obtained as a colourless oil (68 mg, 59%). H NMR (CDCl ): δ
3
(
52 μL, 0.38 mmol, 1.5 eq.) in DCM at 50°C for 16 h. The product
6.70 (d, J=8.2 Hz, 1H), 6.33 (s, 1H), 6.31 (d, J=8.2 Hz, 1H), 5.94–5.85
(m, 2H), 5.62–5.55 (m, 1H), 5.36 (d, J=4.3 Hz, 1H), 5.19–5.05 (m, 2H),
3.84 (s, 3H), 3.82 (s, 3H), 3.82–3.80 (m, 3H), 3.80–3.68 (m, 4H), 3.55–
1
was obtained as a colourless oil (87 mg, 65%). H NMR (CDCl ): δ
6.69 (d, J=8.2 Hz, 1H), 6.33 (s, 1H), 6.25 (d, J=8.1 Hz, 1H), 5.89 (s,
3
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12
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
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These are not the final page numbers!

Full Papers
3
2
1
.46 (m, 1H), 2.91–2.69 (m, 2H), 2.57–2.47 (m, 1H), 2.11 (p, J=7.6 Hz,
016) supplemented with 10% foetal bovine serum (FBS) (Invitrogen
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
13
RES
H). C NMR (CDCl ): δ 151.5, 148.7, 143.2, 140.0, 134.8, 133.1,
10100-147), respectively. MDR cell line NCI/Adr was a gift from
3
[39]
32.3, 129.4, 117.7, 117.2, 112.2, 102.8, 100.9, 85.3, 72.0, 60.1, 59.3,
Prof. Kenneth H. Cowan in the National Cancer Institute. Cells
56.4, 56.2, 50.9 (2×), 40.4, 27.2, 15.4. HR-ESMS calcd. for
C
were cultured in a humidified atmosphere containing 5% CO at
2
+
H
N
2
O S
8
[M+H] 505.1639, found 505.1645.
37
°
C. Before use, cell lines were tested for the presence of
24
29
mycoplasma by using a MycoAlert Mycoplasma detection kit as per
manufacturer’s instructions; cells employed in the assays tested
negative for mycoplasma.
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-(piperidin-1-ylsulfonyl)-5,6,7,8-tetrahydro[1,3]dioxolo
[4,5-g]isoquinoline (15l). Compound 15l was synthesised from 11
4
(
98 mg, 0.25 mmol, 1 eq.) according to General Procedure A, with
Cell-cycle assay: Cells were seeded at 4×10 cells per well in 24-well
DBU (76 μL, 0.76 mmol, 3 eq.) and piperidine-1-sulfonyl chloride
plates for 48 h. Cells were then treated with vehicle control,
noscapine analogues (10 μM), vincristine (100 nM), noscapine
(10 μM) (reference controls) or vehicle control for 18 h. Cells were
washed with phosphate-buffered saline (PBS) and detached using
0.05% trypsin (Invitrogen) for 10 min, post-incubation. Following
deactivation of trypsin, cells were stained with Hoechst 33342
working solution (10 mg/mL) (Invitrogen H3570) for 60 min. The
different stages of the cell proliferation cycle were detected by a
FACSCanto II Fluorescence-Activated Cell Sorting (FACS) analyser
(BD Biosciences, Australia) using the Pacific blue channel (450 nm).
The number of cells present at G , S and G /M phases of the cell
(
53 μL, 0.38 mmol, 1.5 eq.) in DCM at 50°C for 16 h. The product
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
4
4
4
5
5
5
5
5
5
5
5
1
was obtained as a colourless oil (52 mg, 39%). H NMR (CDCl ): δ
6
3
.71–6.65 (m, 1H), 6.32 (s, 1H), 6.27 (d, J=8.2 Hz, 1H), 5.92–5.86 (m,
2H), 5.62 (s, 1H), 5.33 (d, J=4.0 Hz, 1H), 5.17–5.04 (m, 2H), 3.82 (s,
3H), 3.81 (s, 6H), 3.45–3.37 (m, 1H), 3.19–3.00 (m, 4H), 2.84–2.70 (m,
1
3
2
H), 2.61–2.46 (m, 1H), 1.58–1.50 (m, 4H), 1.50–1.42 (m, 2H).
C
NMR (CDCl ): δ 151.5, 148.6, 143.1, 134.0, 134.0, 133.1, 132.2, 129.5,
3
1
17.6, 117.1, 112.2, 102.8, 100.8, 85.5, 71.8, 60.1, 59.2, 56.4, 56.3,
7.0 (2×), +40.4, 27.2, 25.5 (2×), 23.9. HR-ESMS calcd. for
4
C H N O S [M+H] 533.1952, found 533.1954.
2
6
33
2
8
1
2
cycle were obtained as percentage values following data processing
with FlowJo flow cytometry analysis software v10. The percentage
increase in arrested cells by noscapine analogues were calculated
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
thoxy-6-(morpholinosulfonyl)-5,6,7,8-tetrahydro[1,3]dioxolo[4,5-
g]isoquinoline (15m). Compound 15m was synthesised from 11
(99 mg, 0.26 mmol, 1 eq.) according to General Procedure A, with
DBU (76 μL, 0.77 mmol, 3 eq.) and morpholine-4-sulfonyl chloride
with reference to the percentage of cells at the G /M phase in the
2
vehicle control using the following formula: [(%cells in G /M
2
(
noscapine analogues) – (%cells in G /M (vehicle control))/(%cells in
2
(
48 μL, 0.38 mmol, 1.5 eq.) in DMF at 60°C for 16 h. The product
G /M (vehicle control))×100%.
2
1
was obtained as a colourless oil (41 mg, 30%). H NMR (CDCl ): δ
6
3
3
.69 (d, J=8.2 Hz, 1H), 6.33 (s, 1H), 6.24 (d, J=8.2 Hz, 1H), 5.91 (d,
Cell viability assay: MCF-7 and PANC-1 cells were seeded at 5×10
J=1.3 Hz, 1H), 5.90 (d, J=1.3 Hz, 1H), 5.65–5.58 (m, 1H), 5.35 (d, J=
.1 Hz, 1H), 5.17–5.06 (m, 2H), 3.84 (s, 3H), 3.82 (s, 3H), 3.82 (s, 3H),
3.65 (t, J=4.7 Hz, 4H), 3.51–3.40 (m, 1H), 3.24–3.01 (m, 4H), 2.86–
cells per well in 96-well plates 16 h before treatment. Cells were
then treated with various concentration of noscapine analogues.
After 72 h of treatment, cell viability was determined following
incubation with resazurin dye (CellTiter-Blue assay – Promega
G8080). The emitted fluorescence was detected using Envision
microplate reader (PerkinElmer) at 560 nm excitation/590 nm
emission wavelengths. The obtained fluorescence values were
plotted against log concentration of the inhibitor, and EC50 values
were calculated using GraphPad Prism statistical software (v7).
4
13
2
1
1
.73 (m, 2H), 2.61–2.50 (m, 1H). C NMR (CDCl ): δ 151.5, 148.6,
3
43.1, 140.0, 134.0, 133.3, 132.2, 129.8, 117.7, 117.3, 112.2, 102.8,
00.8, 85.8, 71.8, 60.1, 59.2, 56.4, 56.1, 41.9 (2×), 40.4, 27.3, 13.8
+
(2×). HR-ESMS calcd. for C H N O S [M+H] 535.1745, found
25
31
2
9
5
35.1751.
(
R)-5-((S)-4,5-Dimethoxy-1,3-dihydroisobenzofuran-1-yl)-4-me-
RES
thoxy-6-((4-methylpiperazin-1-yl)sulfonyl)-5,6,7,8-tetrahydro[1,3]
dioxolo[4,5-g]isoquinoline (15n). Compound 15n was synthesised
from 11 (95 mg, 0.25 mmol, 1 eq.) according to General Procedure
A, with DBU (73 μL, 0.74 mmol, 3 eq.) and 4-methylpiperazine-1-
Cell antiproliferative assay: MCF-7 and NCI/Adr cells were both
cultured in high-glucose (25 mM) DMEM supplemented with 10%
FBS and 100 U/mL penicillin and 100 μg/mL streptomycin. Every
RES
third passage, the NCI/Adr
cells were treated with 3 μM
sulfonyl chloride (73 mg, 0.37 mmol, 1.5 eq.) in DMF at 60°C for
doxorubicin to maintain selection pressure for the resistance
1
3
1
6 h. The product was obtained as a yellow oil (46 mg, 34%). H
phenotype. Cells were seeded at a density of 3×10 cells per well
NMR (CDCl ): δ 6.69 (d, J=8.2 Hz, 1H), 6.32 (s, J=5.4 Hz, 1H), 6.26
in 96-well plates and recovered in standard culture condition for
48 hours prior to drug treatment. Following the addition of drugs,
cells were incubated for a further 96 h before measuring the
number of live cells using a standard MTT assay. The percentage of
live cells was plotted as a function of drug concentration and fitted
with the general dose response relationship. The extent of cell
death and the drug potency EC50 were estimated from the non-
linear regression using GraphPad Prism (v5). The EC50 values of
noscapine and the derivatives were compared using One-way
ANOVA with the Bonferroni post-hoc test. A value of p<0.05 was
considered statistically significant.
3
(
1
d, J=8.2 Hz, 1H), 5.91–5.85 (m, 2H), 5.61 (s, 1H), 5.34 (d, J=4.0 Hz,
H), 5.18–4.98 (m, 2H), 3.81 (s, 3H), 3.81 (s, 3H), 3.80 (s, 3H), 3.46–
3.36 (m, 1H), 3.29–3.09 (m, 4H), 2.84–2.72 (m, 2H), 2.53 (dt, J=10.4,
13
8.5 Hz, 1H), 2.47–2.36 (m, 4H), 2.27 (s, 3H). C NMR (CDCl
1
1
4
): δ 151.5,
3
48.7, 143.2, 139.9, 134.0, 133.0, 132.2, 129.4, 117.6, 116.9, 112.2,
02.8, 100.9, 85.5, 71.9, 60.1, 59.3, 56.4, 56.3, 54.4 (2×), 46.0 (2×),
+
5.9, 40.4, 27.3. HR-ESMS calcd. for C H N O S [M+H] 548.2061,
26
34
3
8
found 548.2066.
Pharmacology
Tubulin polymerisation assay. Tubulin polymerisation was recorded
turbidimetrically at 340 nm in PerkinElmer EnVision2101 Multilabel
Reader equipped with temperature controllers. Compounds of
interest (2 mM in sterile-filtered DMSO) were dissolved in General
À 2
Compound preparation and storage. A 10 M stock solution was
prepared for each compound by dissolving the compound in
sterile-filtered DMSO (Sigma D2650), and the solution was stored at
À 20°C until use.
Tubulin Buffer (80 mM PIPES pH 6.9, 2 mM MgCl , 0.5 mM EGTA) to
2
a final concentration of 10 μM and were added to 96-half area well
Cell culture and reagents: The human breast cancer MCF-7 (HTB-22)
and pancreatic cancer PANC-1 (CRL-1469) cell lines were purchased
from American Type Culture Collection (ATCC) and cultured in
Minimum Essential Medium alpha (MEMα) (Invitrogen 32561-037)
and Dulbecco’s Modified Eagle Medium (DMEM) (Invitrogen 10566-
plate and kept at 37°C. Tubulin was dissolved to
a final
concentration of 3 mg/mL in 80 mM PIPES pH 6.9, 0.5 mM MgCl ,
2
0
.5 mM EGTA, 1 mM GTP, 10.2% glycerol and was kept at 4°C prior
to addition to plate. Kinetic absorbance reading started immedi-
ChemMedChem 2019, 14, 1–15
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© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
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Full Papers
ately following addition of tubulin to the plate, and was recorded
at 37°C over a period of 60 min. The tubulin polymerisation assay
kit (BK006P) used to conduct this study was supplied by
Cytoskeleton Inc.
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Acknowledgements
The authors thank the Australian government for their support
through an Australian Postgraduate Award to C.Y. The authors
also thank the US National Cancer Institute (NCI) for screening
selected compounds in their NCI-60 Human Tumour Cell Lines
Screen. Thanks also to Professor Colin Pouton for helpful
discussions regarding the biological assays and Sun Pharmaceut-
ical Industries for the kind donation of noscapine.
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Manuscript received: August 15, 2019
Revised manuscript received: October 12, 2019
Version of record online: ■■■, ■■■■
ChemMedChem 2019, 14, 1–15
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© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
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FULL PAPERS
Noscapine displays weak anticancer
efficacy, and numerous research
efforts have attempted to generate
more potent noscapine analogues.
Herein, we report the synthesis and
pharmacological evaluation of a
series of N-sulfonyl and N-sulfamoyl
noscapine derivatives. A number of
these sulfonyl-containing noscapi-
noids demonstrated improved activ-
ities relative to noscapine.
C. Yong, Dr. S. M. Devine, X. Gao, A.
Yan, Prof. R. Callaghan, Dr. B.
Capuano, Prof. P. J. Scammells*
1
– 15
A Novel Class of N-Sulfonyl and N-
Sulfamoyl Noscapine Derivatives
that Promote Mitotic Arrest in
Cancer Cells
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Compound 14q displayed sub-micro-
molar potency against MCF-7, PANC-
1, MDA-MB-435 and SK-MEL-5 cells,
and the antiproliferative effect was
also maintained against drug-
RES
resistant NCI/Adr cells despite high
expression of the multidrug efflux
pump, P-glycoprotein.