Steroidomimetic tetrahydroisoquinolines for the design of new microtubule disruptors

Article abstract of DOI:10.1021/ml200232c

Structure-activity relationship translation offers an expeditious means for discovery of new active series. This approach was applied to discover tetrahydroisoquinoline (THIQ)-based steroidomimetic microtubule disruptors. The two A-ring elements of a thre

Full text of DOI:10.1021/ml200232c

Letter
pubs.acs.org/acsmedchemlett
Steroidomimetic Tetrahydroisoquinolines for the Design of New
Microtubule Disruptors
Mathew P. Leese,† Fabrice Jourdan,† Wolfgang Dohle,† Meriel R. Kimberley,† Mark P. Thomas,†
Ruoli Bai,‡ Ernest Hamel,‡ Eric Ferrandis,§ and Barry V. L. Potter*,†
†Medicinal Chemistry, Department of Pharmacy & Pharmacology, University of Bath, Claverton Down, Bath, BA2 7AY, United
Kingdom
‡Screening Technologies Branch, Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis, National
Cancer Institute at Frederick, National Institutes of Health, Frederick, Maryland 21702, United States
§IPSEN, Institut de Recherche Henri Beaufour, 91966 Les Ulis Cedex, France
S
* Supporting Information
ABSTRACT: Structure−activity relationship translation of-
fers an expeditious means for discovery of new active series.
This approach was applied to discover tetrahydroisoquinoline
(THIQ)-based steroidomimetic microtubule disruptors. The
two A-ring elements of a three-point steroidal pharmacophore
were incorporated into a THIQ-based A,B-ring mimic to
which an H-bond acceptor was attached as the third motif.
Optimization of the representative 6c through conformational
biasing delivered a 10-fold gain in activity and a new series of
microtubule disruptors (e.g., 9c) with antiproliferative activity in the nanomolar range. The THIQ derivatives match, or surpass,
the activities of the steroidal series and exhibit improved physicochemical properties.
KEYWORDS: Tetrahydroisoquinoline, microtubule disruptor, structure−activity relationship, steroidomimetic
espite great promise, exploration of the anticancer
Dapplications of colchicine site binding microtubule
disruptors has yet to deliver an approved drug for clinical
use. In contrast, compounds binding to the vinca and taxane
sites on tubulin have found wide application and are arguably
the most successful class of anticancer agents available.1
Colchicine site binders such as the combretastatins, 2-
methoxyestradiol, and molecules inspired by, or derived from,
these and related compounds do, however, remain in various
stages of development.2 Our interest in this class arose from the
Figure 1. Design of prototypical tetrahydroisoquinoline-based micro-
tubule disruptors by SAR translation.
discovery that sulfamoylation of 2-methoxyestrone yields a
compound that, as well as inhibiting steroid sulfatase, a new
enzyme target for postmenopausal hormone-dependent breast
translating the key elements of the steroidal pharmacophore
cancer,3 displays far higher in vitro and in vivo antiproliferative
into alternate scaffolds. It seemed reasonable to commence by
activity against cancer cells than 2-substituted estradiols.4,5
constructing prototypical systems from appropriately function-
Further studies established that the various anticancer effects of
alized heterocyclic mimics of the A,B-ring system to which
the sulfamoylated 2-substituted estrogen derivatives can be
groups with an appropriate hydrogen bond acceptor to address
ascribed to their ability to disrupt normal microtubule
polymerization6 and that, unlike 2-methoxyestradiol, these
a third key interaction at the colchicine site could be appended.
sulfamates display high oral bioavailability.7 The steroidal
Tetrahydroisoquinoline (THIQ)-based systems appeared ideal
for this purpose since they possess a good degree of the rigidity
present in the steroidal system, the N-2 nitrogen offers an
achiral site of attachment for library generation, and a
derivatives bind to tubulin in a competitive manner versus
colchicine,8 and their activity is independent of estrogen
receptor status.4
Having established the structure−activity relationship for
antiproliferative activity in the sulfamoylated steroidal micro-
tubule disruptor series (Figure 1),5,8−14 we wished to explore
the potential to generate new series of anticancer agents by
Received: April 8, 2011
Accepted: October 31, 2011
Published: October 31, 2011
© 2011 American Chemical Society
5
dx.doi.org/10.1021/ml200232c|ACS Med. Chem. Lett. 2012, 3, 5−9

ACS Medicinal Chemistry Letters
Letter
a
Scheme 1. Synthesis of THIQ-Based Microtubule Disruptors
a
Reagents and conditions: (i) For 2a, MeNO2, HOAc, for 2b EtNO2; NH4OAc, reflux (ii) for 3a Dioxane/DMF, for 3b EtOH; NaBH4, 0 °C (iii)
Raney Ni, NH2NH2·H2O, MeOH, 50 °C (iv) Ac2O, TEA, DCM, 0 °C to rt (v) (CH2O)n, p-TsOH, PhMe, 120 °C (vi) KOH, EtOH/water, reflux
(vii) ArCH2Cl,* TEA, EtOH, 130 °C, microwave (viii) H2, Pd/C, EtOH/THF (ix) H2NSO2Cl, DMA, 0 °C to rt: *R2 a = H, b = 2′-MeO, c = 3′-
MeO, d = 4′-MeO.
convergent, flexible entry to screening candidates could be
envisaged (Figure 1). Furthermore, precedent for the successful
use of tetrahydroisoquinolines in the generation of selective
estrogen receptor modulators was available,15 and we have
recently applied the same template for the generation of
chimeric microtubule disruptors.16 We now describe how this
approach was used to discover a new series of microtubule
disruptors optimized to surpass, in many respects, the steroidal
template that inspired their design.
We required an efficient synthesis of 6-hydroxy-7-methoxy
tetrahydroisoquinoline and selected an approach based on the
Pictet−Spengler reaction (Scheme 1).17,18 Benzyl isovanillin 1
was subjected to a Henry aldol reaction with nitromethane, and
the resultant nitro alkene 2a was reduced in two steps to yield
the corresponding phenethylamine. To achieve good yields
from the key Pictet−Spengler cyclization, it proved expeditious
to acylate the phenethylamine to give 3a, since purification of
the cyclized product 4 was greatly simplified and higher isolated
yields were accordingly obtained. With the tetrahydroisoquino-
line core 4 in hand, conversion to the prototypical screening set
of N-benzyl and 2′-, 3′-, and 4′-methoxybenzyl derivatives was
carried out by cleaving the N-acetyl group and then benzylating
at N-2 with the appropriate benzyl halide. Deprotection of the
6-O-benzyl group gave the phenols 5, followed by sulfamoy-
lation with sulfamoyl chloride in DMA to deliver the desired
putative steroidomimetics 6.
Table 1. Activity of THIQ Derivatives against the
Proliferation of DU-145 Human Prostate Cancer Cells
a
DU-145 GI50
R1
X
R2
R3
R4
(μM)
5a
6a
5b
6b
5c
6c
5d
6d
8a
9a
8b
9b
8c
9c
8d
9d
11
H
H
H
H
H
H
H
H
H
H
H
H
H
H
>100
80.5
>100
7.8
SO2NH2
H
H
H
OMe
OMe
H
SO2NH2
H
H
H
OMe
OMe
H
>100
2.1
SO2NH2
H
H
H
H
H
OMe
OMe
H
>100
57.2
>100
2.2
SO2NH2
H
H
H
H
Me
Me
Me
Me
Me
Me
Me
Me
H
H
SO2NH2
H
H
H
H
OMe
OMe
H
H
H
>100
3.4
SO2NH2
H
H
H
OMe
OMe
H
H
3.73
0.222
>100
1.1
SO2NH2
H
H
H
H
OMe
OMe
SO2NH2
H
H
b
>100
a
b
SE ≤ 7%, n = 3. For the structure of 11, see Scheme 1.
The prototypical library of phenols 5a−d and sulfamates 6a−
d was then screened for antiproliferative activity against DU-
145 prostate cancer cells.14 The results obtained are presented
in Table 1. Phenol derivatives 5a−d show no significant
activity. In contrast, the sulfamates 6a−d display promising
activity, with the 2′- and 3′-methoxybenzyl derivatives 6b and 6c
causing 50% growth inhibition of DU-145 cells at concen-
trations of 7.8 and 2.1 μM, respectively. The unsubstituted
benzyl 6a and 4′-methoxybenzyl 6d compounds are inactive,
and it would appear that the 2′- and 3′-methoxy substituents
best map to the appropriate pharmacophore vector for
antiproliferative effect. To further evaluate this hypothesis, we
synthesized the analogous sulfamate derivative 11 lacking the 7-
methoxy group. THIQ 11 was synthesized in a similar manner
to that outlined for the conversion of 3−6 from commercially
available 3-methoxyphenethylamine 10 with variation only in
the deprotection chemistry applied.19 Screening of 11 for
antiproliferative activity revealed that, like 5a−d and 6a, the
compound is devoid of significant activity, and thus, when any
of the three pharmacophore elements of the design template is
deleted from prototypical lead 6c, so is any antiproliferative
effect. These results validate our hypothesis that translating the
key pharmacophoric elements of the steroidal series into a
nonsteroidal motif could deliver new series of active agents.
6
dx.doi.org/10.1021/ml200232c|ACS Med. Chem. Lett. 2012, 3, 5−9

ACS Medicinal Chemistry Letters
Letter
Clearly, however, the activity of 6c is still an order of
activities of 9a and 9c are analogous to those displayed by 2-
methoxy-17-deoxyestrone-3-O-sulfamate and 2-methoxyes-
trone-3-O-sulfamate in the steroidal series (GI50 values of 2.2
μM and 300 nM in MCF-7 breast cancer cells),5,10 indicating
the complementary nature of the SAR in steroidal and THIQ-
based agents. It seems reasonable to conclude that these THIQ
derivatives are good steroidomimetics and the strategy could
equally be applied to other targets interacting with steroidal
ligands.
With these excellent in vitro data in hand, we wished to
establish the microtubule disruptor activity of 9c, which was
compared with the potent microtubule disruptor combretasta-
tin A-4 (CA-4) and a representative steroid derivative 12a
(Table 2). As can be seen, 9c disrupts the polymerization of
magnitude below that of the parent steroidal compounds.11
A
number of factors could possibly explain this, including the
nonoptimal nature of the methoxy group as an H-bond
acceptor, the methylene group as a linker between the THIQ
core and our D-ring mimic, or indeed a possible intrinsic
reduction of activity due to the presence of the basic amine in
the THIQ core. One potential factor that we were keen to
explore first is the effect of free precession of the N-benzyl
group. This rotational freedom should logically result in only a
small population of conformations that project the H-bond
acceptor at the appropriate vector to mimic the steroidal
compound (Figure 2). We thus examined whether methylation
Table 2. Activity of THIQ Derivative 9c as an Inhibitor of
a
Tubulin Polymerization and Colchicine Binding to Tubulin
inhibition of colchicine
binding
inhibition of tubulin assembly
% inhibition ± SD
compd
IC50 (μM) ± SD
5 μM
50 μM
CA-4
12a
12b
9c
1.3 ± 0.1
2.5 ± 0.1
1.3 ± 0.1
15 ± 1
98 ± 0.6
24.5 ± 5
78 ± 0.9
13 ± 2
74 ± 4
59 ± 2
a
Data for 12b are taken from ref 8.
tubulin with an IC50 of 15 ± 1 μM. Interestingly, this is less
active than the steroidal derivatives with comparable anti-
proliferative activity [e.g., 12a, GI50 (DU-145) 340 nM, IC50
with tubulin 2.5 μM].8 Ultimately, the concentration required
in tubulin-based assays far exceeds the antiproliferative dose
and, most likely, it suffices to disrupt microtubule dynamics to
arrest the cell cycle rather than cause a catastrophic
depolymerization event. It should also be recalled that the
nominal concentration in antiproliferative assays is that added
to the culture medium, rather than the actual concentration of
the agent within the cells. We also determined that 9c inhibits
colchicine binding to tubulin, and its effect is similar to that of
the steroidal derivative 12a, with both agents significantly less
active than CA-4. It thus appears reasonable to suggest that the
interaction of the novel THIQ derivative 9c can at least
partially be ascribed to its ability to disrupt the normal dynamic
polymerization of tubulin by interaction at, or around, the
colchicine site. Combined with our determination that the
pharmacophore elements requisite for activity in the steroidal
series are required for optimal antiproliferative activity in
THIQs both with, and without, a substituent at C-3 these
results serve to validate our design hypothesis.
To shed more light on the mechanism of action of the novel
THIQ derivatives, 6c, 8c, and 9c were evaluated as inhibitors of
the proliferation of human CA46 Burkitt lymphoma cells, along
with CA-4 and the steroidal compound 12b. Because
antitubulin agents classically cause cells to accumulate in
mitosis, the mitotic index with each agent at 10 times the IC50
was also determined. In untreated cells, the mitotic index in
CA46 cells is always 2−4%.20 These data are presented in Table
3. With all three compounds, as well as CA-4 and 12b, a
marked increase in the mitotic index occurred, a strong
indication that at the cellular level the antiproliferative
mechanism involved an interaction with tubulin.
Figure 2. Introduction of C-3 substitution as a means to favor a
“steroid-like” conformation.
of the THIQ core at C-3 could favor the desired “steroid-like”
conformational populations and thus deliver enhanced activity.
A second generation of THIQs 9a−d was synthesized to test
this hypothesis (Scheme 1). Replacement of nitromethane with
nitroethane in the first step of the sequence gave nitroalkene
2b, which was transformed into C-3 methyl THIQ derivatives 8
and 9 in analogy to the synthesis of 5 and 6. THIQs 8a−d and
9a−d were then evaluated as antiproliferative agents and these
data are presented in Table 1.
Comparison of the C-3 methyl derivatives 8a−d and 9a−d
with their unsubstituted analogues 5a−d and 6a−d reveals a
profound difference in antiproliferative activity. As can be seen
8c, in contrast to the C-3 unsubstituted phenols 5a−d, exhibits
activity (3.8 μM) that approaches that of sulfamate 6c (2.1 μM)
and 2-methoxyestradiol (1.2 μM). The activities of the
sulfamoylated compounds are even more encouraging, with a
GI50 of 220 nM obtained for the 3′-methoxybenzyl derivative
9c. A 10-fold gain in activity is thus realized by incorporating a
C-3 methyl group as a conformational bias (cf. 9c and 6c).
Notable too are the activities of 9a and 9d, whose
antiproliferative activities are in the low micromolar range,
whereas the corresponding C-3 unsubstituted derivatives 6a
and 6d show no significant activities at 100 μM. The activities
of 9a (2.2 μM), which lacks the H-bond acceptor, and 9d (1.1
μM), which bears the H-bond acceptor, but apparently not at
the vector to the A-ring pharmacophore elements to confer
antiproliferative activity, suggests that the benzyl group is
picking up positive lipophilic interactions associated with the
steroidal D ring and thus serves to some extent as a D-ring
mimic. Interestingly, in support of this observation, the relative
7
dx.doi.org/10.1021/ml200232c|ACS Med. Chem. Lett. 2012, 3, 5−9

ACS Medicinal Chemistry Letters
Letter
material is available free of charge via the Internet at http://
pubs.acs.org.
Table 3. Activity of Compounds against the Proliferation of
Human CA46 Burkitt Lymphoma Cells
compd
IC50 (nM) ± SD
% mitotic cells at 10 × IC50
AUTHOR INFORMATION
■
CA-4
12b
6c
6 ± 1
68 ± 8
60 ± 10
66 ± 6
45 ± 10
71 ± 7
350 ± 100
2300 ± 300
2500 ± 700
230 ± 100
Corresponding Author
*Tel: +44 1225 386639. Fax: +44 1225 386114. E-mail: B.V.L.
Potter@bath.ac.uk.
8c
9c
Funding
This work was supported by Sterix Ltd., a member of the Ipsen
group, by VIP awards and a Programme Grant (082837) from
the Wellcome Trust.
The effects of C-3 methylation on the energy states of
conformers of a model system, 3-methyl-N-benzyl-1,2,3,4-
tetrahydroisoquinoline, arising from rotation of the N-benzyl
group, and precession of the phenyl ring of this group around
its axis, were assessed using computational energy calcula-
tions.21 These calculations indicate, as we had postulated, that
in the minimal energy conformation the N-benzyl group is
projected into the area of space close to that occupied by the
steroidal D ring in the estratriene series. This is illustrated in
Figure 3 where the minimal energy state of the (R)-enantiomer
REFERENCES
■
(1) Jordan, M. A.; Wilson, L. Microtubules as a target for anticancer
drugs. Nat. Rev. Cancer 2004, 4, 253−265.
(2) Hadfield, J. A.; Ducki, S.; Hirst, N.; McGown, A. T.; Meijer, L.;
Jezequel, A.; Roberge, M. Progress in Cell Cycle Research. Prog. Cell
Cycle Res. 2003, 309−325.
(3) Reed, M. J.; Purohit, A.; Newman, S. P.; Potter, B. V. L. Steroid
sulfatase: molecular biology, regulation, and inhibition. Endocr. Rev.
2005, 26, 171−202.
(4) Purohit, A.; Hejaz, H. A. M.; Walden, L.; MacCarthy-Morrogh,
L.; Packham, G.; Potter, B. V. L; Reed, M. J. The effect of 2-
methoxyoestrone-3-O-sulphamate on the growth of breast cancer cells
and induced mammary tumours. Int. J. Cancer 2000, 85, 584−589.
(5) Leese, M. P.; Hejaz, H. A. M.; Mahon, M. F.; Newman, S. P.;
Purohit, A.; Reed, M. J.; Potter, B. V. L. A-ring-substituted estrogen-3-
O-sulfamates: potent multitargeted anticancer agents. J. Med. Chem.
2005, 48, 5243−5256.
(6) MacCarthy-Morrogh, L.; Townsend, P. A.; Purohit, A.; Hejaz, H.
A. M.; Potter, B. V. L; Reed, M. J.; Packham, G. Differential effects of
estrone and estrone-3-O-sulfamate derivatives on mitotic arrest,
apoptosis, and microtubule assembly in human breast cancer cells.
Cancer Res. 2000, 60, 5441−5450.
(7) Ireson, C. R.; Chander, S. K.; Purohit, A.; Perera, S.; Newman, S.
P.; Parish, D.; Leese, M. P.; Smith, A. C.; Potter, B. V. L; Reed, R. J.
Pharmacokinetics and efficacy of 2-methoxyoestradiol and 2-
methoxyoestradiol-bis-sulphamate in vivo in rodents. Br. J. Cancer
2004, 90, 932−937.
(8) Jourdan, F.; Leese, M. P.; Dohle, W.; Hamel, E.; Ferrandis, E.;
Newman, S. P.; Purohit, A.; Reed, M. J.; Potter, B. V. L. Synthesis,
Antitubulin, and Antiproliferative SAR of Analogues of 2-Methox-
yestradiol-3,17-O,O-bis-sulfamate. J. Med. Chem. 2010, 53, 2942−2951.
(9) Leese, M. P.; Newman, S. P.; Purohit, A.; Reed, M. J.; Potter, B.
V. L. 2-Alkylsulfanyl estrogen derivatives: synthesis of a novel class of
multi-targeted anti-tumour agents. Bioorg. Med. Chem. Lett. 2004, 14,
3135−3138.
(10) Leese, M. P.; Leblond, B.; Newman, S. P.; Purohit, A.; Reed, M.
J.; Potter, B. V. L. Anti-cancer activities of novel D-ring modified 2-
substituted estrogen-3-O-sulfamates. J. Steroid Biochem. Mol. Biol.
2005, 94, 239−251.
(11) Leese, M. P.; Leblond, B.; Smith, A.; Newman, S. P.; Di Fiore,
A.; De Simone, G.; Supuran, C. T.; Purohit, A.; Reed, M. J.; Potter, B.
V. L. 2-Substituted estradiol bis-sulfamates, multitargeted antitumor
agents: synthesis, in vitro SAR, protein crystallography, and in vivo
activity. J. Med. Chem. 2006, 49, 7683−7696.
(12) Leese, M. P.; Jourdan, F. L.; Gaukroger, K.; Mahon, M. F.;
Newman, S. P.; Foster, P. A.; Stengel, C.; Regis-Lydi, S.; Ferrandis, E.;
Di Fiore, A.; De Simone, G.; Supuran, C. T.; Purohit, A.; Reed, M. J.;
Potter, B. V. L. Structure−activity relationships of C-17 cyano-
substituted estratrienes as anticancer agents. J. Med. Chem. 2008, 51,
1295−1308.
(13) Jourdan, F.; Bubert, C.; Leese, M. P.; Smith, A.; Ferrandis, E.;
Regis-Lydi, S.; Newman, S. P.; Purohit, A.; Reed, M. J.; Potter, B. V. L.
Effects of C-17 heterocyclic substituents on the anticancer activity of
2-ethylestra-1,3,5(10)-triene-3-O-sulfamates: synthesis, in vitro evalua-
Figure 3. Overlay of the minimum energy conformation of 3(R)-
methyl-2-benzyl-1,2,3,4-tetrahydroisoquinoline (cyan) with estradiol
(green) viewed from two perspectives.
is overlaid with the energy-minimized estradiol core (see the
Supporting Information for further detail). In contrast, in the
maximum energy conformation the N-benzyl group eclipses the
C-3 methyl group and is thus far away from the area of space
occupied by the steroidal D ring.21 The energy difference
between these states for either C-3 enantiomer is more than
120 kJ/mol. These calculations support our postulate that C-3
methylation favors adoption of a “steroid-like” conformation,
and thus, it seems reasonable to propose that the positive
effects of C-3 methylation on activity can, to some degree, be
ascribed to this conformational biasing.
We have thus demonstrated how translation of pharmaco-
phore elements can be successfully used to “series hop” and
deliver new series of microtubule disruptors with excellent
antiproliferative activity. These novel THIQ derivatives were
optimized by biasing the conformational population through
introduction of a steric buttress at C-3. In addition to an activity
profile that matches the steroidal compounds from which their
design was inspired, the new THIQ derivatives have low
molecular weight and an excellent solubility profile. They form
water-soluble salts and can be formulated as solutions in mild
aqueous acid. We are actively exploring further optimization
and preclinical development of this series and the application of
THIQ-based steroidomimetics to other therapeutic targets.
ASSOCIATED CONTENT
■
S
* Supporting Information
Experimental procedures and analytical data for compounds 8c
and 9c and details of computational energy calculations. This
8
dx.doi.org/10.1021/ml200232c|ACS Med. Chem. Lett. 2012, 3, 5−9

ACS Medicinal Chemistry Letters
Letter
tion and computational modelling. Org. Biomol. Chem. 2008, 6, 4108−
4119.
(14) Bubert, C.; Leese, M. P.; Mahon, M. F.; Ferrandis, E.; Regis-
Lydi, S.; Kasprzyk, P. G.; Newman, S. P.; Ho, Y. T.; Purohit, A.; Reed,
M. J.; Potter, B. V. L. 3,17-Disubstituted 2-alkylestra-1,3,5(10)-trien-3-
ol derivatives: synthesis, in vitro and in vivo anticancer activity. J. Med.
Chem. 2007, 50, 4431−4443.
(15) Chesworth, R.; Zawistoski, M. P.; Lefker, B. A.; Cameron, K.
O.; Day, R. F.; Mangano, M.; Rosati, R. L.; Colella, S.; Petersen, D. N.;
Brault, A.; Lu, B. H.; Pan, L. C.; Perry, P.; Ng, O.; Castleberry, T. A.;
Owen, T. A.; Brown, T. A.; Thompson, D. D.; DaSilva-Jardine, P.
Tetrahydroisoquinolines as subtype selective estrogen agonists/
antagonists. Bioorg. Med. Chem. Lett. 2004, 14, 2729−2733.
(16) Leese, M. P.; Jourdan, F.; Kimberley, M. R.; Cozier, G. E.;
Thiyagarajan, N.; Stengel, C.; Regis-Lydi, S.; Foster, P. A.; Newman, S.
P.; Acharya, K. R.; Ferrandis, E.; Purohit, A.; Reed, M. J.; Potter, B. V.
L. Chimeric microtubule disruptors. Chem. Comm. 2010, 46, 2907−
2909.
(17) Cox, E. D.; Cook, J. M. The Pictet−Spengler condensation: a
new direction for an old reaction. Chem. Rev. 1995, 95, 1797−1842.
(18) Schlosser, M.; Simig, G.; Geneste, H. Three complementary
methods offering access to 5-substituted 1,2,3,4-tetrahydroisoquino-
lines. Tetrahedron 1998, 54, 9023−9032.
(19) Zhong, H. M.; Villani, F. J.; Marzouq, R. Improved and Practical
Synthesis of 6-Methoxy-1,2,3,4-tetrahydroisoquinoline Hydrochloride.
Org. Process Res. Dev. 2007, 11, 463−465.
(20) Cruz-Monserrate, Z.; Vervoort, H. C.; Bai, R. L.; Newman, D.
J.; Howell, S. B.; Los, G.; Mullaney, J. T.; Williams, M. D.; Pettit, G. R.;
Fenical, W.; Hamel, E. Diazonamide A and a Synthetic Structural
Analog: Disruptive Effects on Mitosis and Cellular Microtubules and
Analysis of Their Interactions with Tubulin. Mol. Pharmacol. 2003, 63,
1273−1280.
(21) See the Supporting Information for full details.
9
dx.doi.org/10.1021/ml200232c|ACS Med. Chem. Lett. 2012, 3, 5−9