Development of highly cytotoxic and actin-depolymerizing biotin derivatives of aplyronine A

Article abstract of DOI:10.1002/anie.201103802

Tied up: A PEG-linked biotin derivative of marine macrolide aplyronine A (ApA; see scheme) is shown to exhibit potent cytotoxicity and cause actin disassembly in tumor cells. This method of introducing a PEG linker at the end of the aliphatic tail should offer perspectives for developing and using versatile actin-targeting molecular probes. PEG=poly(ethylene glycol) Copyright

Full text of DOI:10.1002/anie.201103802

DOI: 10.1002/anie.201103802
Natural Products
Development of Highly Cytotoxic and Actin-Depolymerizing Biotin
Derivatives of Aplyronine A**
Masaki Kita,* Yuichiro Hirayama, Miyuki Sugiyama, and Hideo Kigoshi*
Aplyronine A (ApA, 1), a 24-membered macrolide isolated
from the sea hare Aplysia kurodai, exhibits potent antitumor
activities in vivo against P388 murine leukemia (T/C 545%,
0.08 mgkgÀ1) and several cancers.[1,2] This macrolide has been
shown to depolymerize fibrous actin (F-actin) and inhibit the
polymerization of actin by forming a 1:1 complex with the
monomeric globular molecule (G-actin, Kd = 100 nm).[3] Actin
is one of the most abundant proteins in the cytoskeleton and is
essential for the regulation of various functions, such as
muscle contraction, cell motility, and cell division. Recently,
various actin-depolymerizing agents have been found in
marine invertebrates,[4] and some compounds, such as ula-
pualides,[5] mycalolides,[6] kabiramides,[7] sphinxolides/reidi-
spongiolides,[8] swinholides,[9] and bistramides,[10] which are
similar to 1, show extremely strong cytotoxicities. However,
when comparing the amount of abundant actin molecules
with the amount of 1 incorporated into the cells, the
significant antitumor activities of 1 may not be accounted
for only by its F-actin-severing properties. The modes of
action and the target proteins of ApA and related actin-
targeting natural products in tumor cells have rarely been
clarified, despite their great potential as preclinical candidates
for use in cancer chemotherapy.[11,12]
Figure 1. a) Structure of 1 and b) binding of 1 to actin (PDB code:
1wua). Hydrophobic amino acid residues (T, Y, M, L) at the binding
site of actin are shown in color. L=leucine, M=methionine, T=
threonine, Y=tyrosine.
The structure of aplyronine A can be divided into two
characteristic parts: the C1–C23 macrolactone and the
C24–C34 aliphatic tail (Figure 1a). Structure–activity rela-
tionship studies[13] and photoaffinity labeling experiments[14]
have established that the tail part of 1 is important for potent
actin-depolymerizing activity. In fact, the C21–C34 synthetic
analogue of aplyronine A specifically binds to actin at the
same position as 1. Meanwhile, the functional groups of 1 that
were important for cytotoxicity were found to be the
trimethylserine group on C7, the hydroxy group on C9, and
a conjugated diene moiety on the macrolactone ring.
Recently, an X-ray analysis of the actin/aplyronine A
complex was performed at a resolution of 1.45 ꢀ, and showed
that the tail of 1 intercalates into the hydrophobic cleft
between subdomains (SD) 1 and 3 of actin (Figure 1b).[15] The
amino acid residues of actin, such as Y133, Y143, T148, Y169,
L346, L349, T351, and M355, participate in the hydrophobic
interaction with 1. In contrast, the terminal N-formyl enamide
moiety of 1 is located in a very hydrophilic environment and
interacts with the water molecules inside the actin. The
complex structures of actin with other macrolides, such as
kabiramide C,[16] sphinxolide B,[17] reidispongiolides A and
C,[17] and swinholide A,[18] as well as a polyketide bistra-
mide A[19] have also been reported. The contacts between
actin and each macrolide were similar to those observed in the
actin/aplyronine A complex. On the other hand, the trime-
thylserine moiety of aplyronine A, which is a unique func-
tional group among actin-binding macrolides, and is also
important for cytotoxicity, protrudes from the binding posi-
tion of the macrolactone ring of 1 toward the bulk solvent.
This molecular arrangement implies that binding protein(s)
(other than actin) would be able to attach to this moiety of 1
in the actin/aplyronine A complex and thus may contribute to
the potent antitumor effect of aplyronine A. To clarify this
hypothesis, we have performed chemical and biological
studies on aplyronine A and related actin-targeting macro-
lides.[20] Herein we describe the synthesis and biological
activities of aplyronine A derivatives that possess a biotin
[*] Dr. M. Kita,[+] Y. Hirayama, M. Sugiyama, Prof. Dr. H. Kigoshi[+]
Graduate School of Pure and Applied Sciences
University of Tsukuba
1-1-1 Tennodai, Tsukuba 305-8571 (Japan)
E-mail: mkita@chem.tsukuba.ac.jp
kigoshi@chem.tsukuba.ac.jp
[+] These authors contributed equally to this work.
[**] Support was provided by JSPS through Grants-in-Aid for Scientific
Research (21681028 and 21651091 for M.K., and 20310129 and
23102014 for H.K.), the Kato Memorial Bioscience Foundation, the
Naito Foundation, the Uehara Memorial Foundation, and the
Takeda Science Foundation. We thank Dr. Y. Miwa (University of
Tsukuba) for valuable suggestions.
Supporting information for this article is available on the WWW
under http://dx.doi.org/10.1002/anie.201103802.
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9871

Communications
Table 1: Biological activities of biotinylated ApA derivatives.
moiety, as well as an investigation of their target proteins in
tumor cells.
It has been suggested that the N-formyl enamide moiety,
Cytotoxicity (HeLa S3)
Actin-depolymerizing
IC50 [nm]
activity[a] EC50 [mm]
which is common to various actin-depolymerizing macrolides,
is essential for their potent cytotoxicities.[2,4a,13] However,
because there is a cavity structure inside the 1,3-cleft of the
actin molecule (Figure 1), we anticipated that ApA could be
modified at C34 without a significant loss of activity by
elongating a tethered biotin moiety with hydrophilic groups.
Acidic hydrolysis of the N-formyl enamide moiety of natural
aplyronine A (1)[1i] gave aldehyde 2 (Scheme 1). Reductive
aminoalkylation of 2 with biocytin (5) gave ApA derivative 3.
Similarly, condensation of 2 with a PEG-linked biotin
hydrazide gave another derivative (4).[21] For comparison,
PEG-linked biotin analogue 6 was also prepared from
3-phenylpropionaldehyde.
ApA (1)
0.010
15
0.096
>10000
>10000
1.6
2.1
1.8
>100
>100
3
4
5
6
[a] Activity was monitored by measuring the fluorescence intensity of
pyrenyl actin. Values indicate the concentrations required to depoly-
merize F-actin (3 mm) to 50% of its control amplitude.
Figure 2. Confocal fluorescence images of HeLa S3 cells treated with 1
and 4 for 2 h. a–c) Cells were immunostained with anti-b-tubulin
antibody (green), and co-stained with DAPI (blue) and rhodamine-
phalloidin (red). Scale bars=20 mm. d–g) Monochroic and bright-field
images of the cells in (c). Cells were also stained with Alexa Fluor 647
streptavidin (yellow, pseudo color), and four colors (BGRY) were
merged in (g). Scale bar=20 mm. DAPI=4’,6-diamidino-2-phenylin-
dole, DIC=differential interference contrast, DMSO=dimethyl-
sulfoxide.
compound 4 that had been incorporated into the cells,
intracellular biotin was visualized with fluorescently labeled
streptavidin. In fact, most of the F-actin and biotin in the
cytoplasm were differentially localized (Figure 2d–g). These
significant morphological changes in HeLa S3 cells strongly
indicated that compound 4 may be distributed in the
cytoplasm as a 1:1 complex with G-actin after the rapid
disassembly of multiple filaments. Thus, the actin-severing
mechanism and intracellular behaviour of 4 may be identical
to that of natural 1.
The biotinylated ApA derivatives were then examined to
determine their specificities for binding to actin. When the
binding proteins were competitively eluted by an excess of 1,
only the actins that were attached to biotinylated ApA
analogues 3 or 4 were eluted (Figure 3a). In contrast, actins
were eluted in all lanes by boiling the resins in SDS buffer
(Figure 3b). Thus, considerable amounts of actins were
nonspecifically absorbed to the hydrophobic agarose resin
because of the presence of the lipophilic parts of the actin
molecule (i.e., the 1,3-cleft). Despite this behaviour, resin-
bound compounds 3 and 4 were found to exhibit high affinity
for actin and to have the same binding properties as 1.
Scheme 1. Synthesis of biotinylated ApA derivatives. a) 2m HCl/1,4-
dioxane=1:3, 508C, 80 min; b) 5, MeOH/H2O=1:1, RT, 1 h;
NaBH3CN, RT, 36 h, 24% from 1; c) EZ-Link biotin-PEG4-hydrazide,
MeOH/AcOH=4:1, RT, 48 h, 38% from 1.
The cytotoxicities and in vitro actin-depolymerizing activ-
ities of ApA analogues and model compounds were com-
pared to those of 1 (Table 1). The biocytin-bearing analogue 3
was found to be 1500 times less cytotoxic than 1,[22] whereas
the PEG-linked analogue 4 was only approximately 10 times
less active than 1 (IC50 96 pm). Meanwhile, analogues 3 and 4
showed potent actin-depolymerizing properties, which were
comparable to those of natural ApA. In contrast, model
compounds 5 and 6, which lack the whole parts of 1, did not
exhibit either cytotoxicity or actin-depolymerizing properties.
We also examined the ability of biotinylated ApA
analogue 4 to depolymerize F-actin in living cells (Figure 2).
After treatment with 4 (5 mm) for 2 h, actin disassembly and
the formation of bubble-like blebs in the cytoplasm were
observed, as with 1 (Figure 2b,c). For the detection of
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treatment with 4, a weak band corresponding to actin was
observed, along with nonspecific bands (120 and 74–72 kDa,
biotin carboxylase). In the same experiments, biotin-contain-
ing compounds, namely 4 and/or compounds derived from 4,
were detected in the region of the lowest molecular weight.
Meanwhile, the amounts of bound actins that were eluted by
boiling of the resins were mostly the same as those with ApA
elution (Figure 4a, lanes 2 and 4). These results suggested
that most of the actins bound to 4 were competitively eluted
by ApA, and that the ApA analogue 4 was noncovalently but
highly specifically bound to actin, as in an actin-binding assay.
In addition, two proteins with molecular masses of 40 and
47 kDa were purified with abundant actin from the lysate of
HeLa S3 cells by using a PEG-linked biotinylated ApA
analogue 4 (Figure 4a, lane 4), but not when a biocytinylated
ApA analogue 3 or model compounds 5 and 6 were used.[23]
By a detailed MS/MS analysis of the tryptic peptide fragments
obtained by the in-gel digestion, we confirmed that the 40 and
47 kDa proteins were actin-related proteins 2 and 3 (Arp2
and Arp3), respectively. Notably, as with actin, both Arp2 and
Arp3 were also competitively eluted from the resin by 1
(Figure 4a, lane 2), a result that was established by immuno-
blot analysis (Figure 4e,f). These results suggested the
presence of specific interactions between ApA and actin-
related proteins. Arp2 and Arp3 are the key proteins of the
Arp2/3 complex, which binds to the sites of an existing actin
filament and initiates growth of a new actin filament to form
branched-actin-filament networks.[24,25] The amino acid
sequences and three-dimensional structures of Arp2 and
Arp3 are highly similar (46% and 36% identity) to those of
monomeric actin.[26,27] Moreover, the remaining components
of the Arp2/3 complex, ARPC1–5 (16–40 kDa), were not
purified by the pull-down assay with 4. Thus, it was suggested
that ApA (1) may bind to Arp2 or Arp3 to give 1:1 complexes,
as with the actin/aplyronine A complex, and inhibit the ability
of the Arp2/3 complex to bind to and branch F-actin.[28] These
properties may help to enhance the potent actin filament
disassembly caused by 1.
Figure 3. Interaction of biotinylated ApA derivatives with actin. Actin in
G buffer was treated with compounds 3–6 or DMSO (shown as À)
that were preloaded on NeutrAvidin agarose. Binding actin was eluted
by a) treatment with 1 (5 mm) or by b) boiling in SDS buffer, and
detected with silver stain.
Furthermore, we investigated the interactions of biotin-
ylated ApA analogue 4 with whole cellular proteins. To
minimize interference with intrinsic biotin-binding molecules,
an excess amount of 4 was pretreated with NeutrAvidin
agarose, on which the lysate of HeLa S3 cells was loaded.
When the binding proteins were eluted by boiling in SDS
buffer, two bands with molecular masses of 43 and 200 kDa
were clearly detected along with several nonspecific bands
(Figure 4a, lane 4). Through the use of peptide mass finger-
printing (PMF) and immunoblot analyses, the two main bands
were identified to be b actin and nonmuscle myosin II (heavy
chain), respectively (Figure 4a,d).[23] Myosin II is an F-actin
binding protein with ATPase activity. However, no significant
differences in the staining patterns of myosin II were
observed between 4 and other biotinylated compounds 3, 5,
and 6.[23] Thus, myosin II may bind not directly to the ApA
derivatives, but rather to the abundant actin molecules that
were trapped on the resin directly or through biotinylated
compounds.
To confirm the interactions of 4 and actin, biotinylated
proteins were purified by a pull-down assay and subsequently
detected by immunoblotting analysis using a horseradish
peroxidase (HRP) conjugated streptavidin (Figure 4b). Upon
Two small aromatic molecules, CK-636 and CK-548, were
recently discovered and were shown to bind to the Arp2/3
complex and inhibit its ability to nucleate actin filaments.[29]
Molecules CK-636 and CK-548 bind between Arp2 and Arp3
or insert into the hydrophobic core of Arp3 in the Arp2/3
complex, but do not depolymerize F-actin. Meanwhile, to the
best of our knowledge, this is the first example in which actin-
depolymerizing molecules can also bind to actin-related
proteins.
In summary, highly bioactive biotinylated derivatives of
aplyronine A (1) have been developed. Since the N-formyl
enamide moiety in 1 is a conserved structure on various actin-
depolymerizing marine macrolides, this method of introduc-
ing a PEG linker at the end of the aliphatic tail should offer
perspectives for developing and using versatile actin-targeting
molecular probes.[30] Further structural and functional anal-
yses of ApA-binding proteins as well as their roles in the
antitumor activities of 1 are currently underway.
Figure 4. Pull-down assay of ApA-binding proteins in cell lysate.
a) HeLa S3 cell lysate (lane 1) was treated with 4 (probe +) or DMSO
(probe À), which were preloaded on NeutrAvidin agarose. Binding
proteins were eluted by treatment with 1 (5 mm) or by boiling in SDS
buffer, subjected to 10% SDS-PAGE, and detected with silver stain.
Arrowheads indicate Arp2 and Arp3. b–f) Immunoblotting analysis.
Proteins in lanes 4 and 5 in (a) were detected with b) HRP-conjugated
streptavidin, c) anti-myosin II, or d) anti-b-actin, respectively. Similarly,
proteins in lanes 2 and 3 in (a) were detected with e) anti-Arp2 and
anti-Arp3 (1:1 mix) or f) anti-ACTR3.
Received: June 4, 2011
Revised: August 4, 2011
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9873

Communications
Chem. 2003, 3, 593; b) J. R. Peterson, T. J. Mitchison, Chem.
Biol. 2002, 9, 1275.
[12] For reviews on cancer chemotheraphy, see: a) I. Spector, F.
Braet, N. R. Shochet, M. R. Bubb, Microsc. Res. Tech. 1999, 47,
18; b) A. Giganti, E. Friederich, Prog. Cell Cycle Res. 2003, 5,
511.
[13] a) K. Suenaga, N. Kamei, Y. Okugawa, M. Takagi, A. Akao, H.
Kigoshi, K. Yamada, Bioorg. Med. Chem. Lett. 1997, 7, 269;
b) H. Kigoshi, K. Suenaga, M. Takagi, A. Akao, K. Kanematsu,
N. Kamei, Y. Okugawa, K. Yamada, Tetrahedron 2002, 58, 1075.
[14] T. Kuroda, K. Suenaga, A. Sakakura, T. Handa, K. Okamoto, H.
Kigoshi, Bioconjugate Chem. 2006, 17, 524.
[15] K. Hirata, S. Muraoka, K. Suenaga, K. Kuroda, K. Kato, H.
Tanaka, M. Yamamoto, M. Takata, K. Yamada, H. Kigoshi,
J. Mol. Biol. 2006, 356, 945.
[16] V. A. Klenchin, J. S. Allingham, R. King, J. Tanaka, G. Marriott,
I. Rayment, Nat. Struct. Biol. 2003, 10, 1058.
[17] J. S. Allingham, A. Zampella, M. V. DꢁAuria, I. Rayment, Proc.
Natl. Acad. Sci. USA 2005, 102, 14527.
[18] V. A. Klenchin, R. King, J. Tanaka, G. Marriott, I. Rayment,
Chem. Biol. 2005, 12, 287.
[19] S. A. Rizvi, V. Tereshko, A. A. Kossiakoff, S. A. Kozmin, J. Am.
Chem. Soc. 2006, 128, 3882.
Keywords: actin depolymerization · antitumor agents ·
chemical probes · natural products · proteins
.
[1] For the isolation and structures of aplyronines, see: a) K.
Yamada, M. Ojika, T. Ishigaki, Y. Yoshida, H. Ekimoto, M.
Arakawa, J. Am. Chem. Soc. 1993, 115, 11020; b) M. Ojika, H.
Kigoshi, T. Ishigaki, I. Tsukada, T. Tsuboi, T. Ogawa, K.
Yamada, J. Am. Chem. Soc. 1994, 116, 7441; c) K. Suenaga, T.
Ishigaki, A. Sakakura, H. Kigoshi, K. Yamada, Tetrahedron Lett.
1995, 36, 5053; d) M. Ojika, H. Kigoshi, T. Ishigaki, K. Yamada,
Tetrahedron Lett. 1993, 34, 8501; e) M. Ojika, H. Kigoshi, T.
Ishigaki, M. Nishiwaki, I. Tsukada, K. Mizuta, K. Yamada,
Tetrahedron Lett. 1993, 34, 8505; for the synthesis of aplyronines,
see: f) H. Kigoshi, M. Ojika, T. Ishigaki, K. Suenaga, T. Mutou,
A. Sakakura, T. Ogawa, K. Yamada, J. Am. Chem. Soc. 1994,
116, 7443; g) H. Kigoshi, M. Ojika, K. Suenaga, T. Mutou, J.
Hirano, A. Sakakura, T. Ogawa, M. Nisiwaki, K. Yamada,
Tetrahedron Lett. 1994, 35, 1247; h) H. Kigoshi, K. Suenaga, T.
Mutou, T. Ishigaki, T. Atsumi, H. Ishiwata, A. Sakakura, T.
Ogawa, M. Ojika, K. Yamada, J. Org. Chem. 1996, 61, 5326; i) M.
Ojika, H. Kigoshi, Y. Yoshida, T. Ishigaki, M. Nishiwaki, I.
Tsukada, M. Arakawa, H. Ekimoto, K. Yamada, Tetrahedron
2007, 63, 3138.
[2] For reviews, see: a) K. Yamada, M. Ojika, H. Kigoshi, K.
Suenaga, Nat. Prod. Rep. 2009, 26, 27; b) K. Yamada, M. Ojika,
H. Kigoshi, K. Suenaga, Proc. Jpn. Acad. Ser. B 2010, 86, 176;
c) K. Suenaga, H. Kigoshi, J. Synth. Org. Chem. Jpn. 2006, 64,
1273 (in Japanese).
[3] S. Saito, S. Watabe, H. Ozaki, H. Kigoshi, K. Yamada, N.
Fusetani, H. Karaki, J. Biochem. (Tokyo) 1996, 120, 552.
[4] For general reviews on actin-depolymerizing natural products,
see: a) J. S. Allingham, V. A. Klenchin, I. Rayment, Cell. Mol.
Life Sci. 2006, 63, 2119; b) K. S. Yeung, I. Paterson, Angew.
Chem. 2002, 114, 4826; Angew. Chem. Int. Ed. 2002, 41, 4632, and
references therein.
[5] E. Vincent, J. Saxton, C. Baker-Glenn, I. Moal, J. D. Hirst, G.
Pattenden, P. E. Shaw, Cell. Mol. Life Sci. 2007, 64, 487.
[6] a) M. Hori, S. Saito, Y. Z. Shin, H. Ozaki, N. Fusetani, H. Karaki,
FEBS Lett. 1993, 322, 151; b) S. Saito, S. Watabe, H. Ozaki, N.
Fusetani, H. Karaki, J. Biol. Chem. 1994, 269, 29710; c) S. Wada,
S. Matsunaga, S. Saito, N. Fusetani, S. Watabe, J. Biochem. 1998,
123, 946.
[7] a) J. Tanaka, Y. Yan, J. Choi, J. Bai, V. A. Klenchin, I. Rayment,
Proc. Natl. Acad. Sci. USA 2003, 100, 13851; b) C. Petchprayoon,
K. Suwanborirux, J. Tanaka, Y. Yan, T. Sakata, G. Marriott,
Bioconjugate Chem. 2005, 16, 1382.
[8] a) X. Zhang, L. Minale, A. Zampella, C. D. Smith, Cancer Res.
1997, 57, 3751; b) R. D. Perrins, G. Cecere, I. Paterson, G.
Marriott, Chem. Biol. 2008, 15, 287.
[9] a) M. R. Bubb, I. Spector, A. D. Bershadsky, E. D. Korn, J. Biol.
Chem. 1995, 270, 3463; b) S. Saito, S. Watabe, H. Ozaki, M.
Jobayashi, T. Suzuki, H. Kobayashi, N. Fusetani, H. Karaki,
J. Biochem. (Tokyo) 1998, 123, 571.
[20] M. Kita, H. Watanabe, T. Ishitsuka, Y. Mogi, H. Kigoshi,
Tetrahedron Lett. 2010, 51, 4882, and references therein.
[21] Stereoisomers of the imines in 4 and 6 were not separable (E/Z =
2:1), and therefore they were used as mixtures in biochemical
studies.
[22] Biocytinylated compound 3 bears a carboxyl group, which may
influence its ability to permeate into living cells or give it lower
affinity for targeted molecules, thus causing a moderate decrease
in cytotoxicity.
[23] See the Supporting Information for details.
[24] a) R. D. Mullins, J. A. Heuser, T. D. Pollard, Proc. Natl. Acad.
Sci. USA 1998, 95, 6181; b) L. Blanchoin, K. J. Amann, H. N.
Higgs, J. B. Marchand, D. A. Kaiser, T. D. Pollard, Nature 2000,
404, 1007.
[25] For a review on the Arp2/3 complex, see: E. D. Goley, M. D.
Welch, Nat. Rev. Mol. Cell Biol. 2006, 7, 713.
[26] R. C. Robinson, K. Turbedsky, D. A. Kaiser, J. B. Marchand,
H. N. Higgs, S. Choe, T. D. Pollard, Science 2001, 294, 1679.
[27] The hydrophobic cleft between SD1 and SD3 of actin, to which 1
binds, includes the helixes aD and aK and the sheets b10–b12.
These secondary structures are highly conserved among actin,
Arp2, and Arp3. See the Supporting Information for details.
[28] The interactions of ApA with Arp2/Arp3 were shown only by
pull-down experiments at this stage, and we could not eliminate
the possibility that ApA targets both actin and the Arp2/3
complex to form the ternary complex. We hope to verify this
behaviour in future studies.
[29] B. J. Nolen, N. Tomasevic, A. Russell, D. W. Pierce, Z. Jia, C. D.
McCormick, J. Hartman, R. Sakowicz, T. D. Pollard, Nature
2009, 460, 1031.
[30] There is one report on the biocytinylated mycalolide and
kabiramide derivatives (see Ref. [6c]). However, the cytotox-
icities and actin-depolymerizing activities of these compounds
have not been described. Biocytinylated mycalolide B was
shown to bind to some cellular proteins including actin, but its
binding properties were nonspecific because of the formation of
covalent bonds at the unsaturated ketone moiety.
[10] a) A. V. Statsuk, R. Bai, J. L. Baryza, V. A. Verma, E. Hamel,
P. A. Wender, S. A. Kozmin, Nat. Chem. Biol. 2005, 1, 383;
b) S. A. Rizvi, D. S. Courson, V. A. Keller, R. S. Rock, S. A.
Kozmin, Proc. Natl. Acad. Sci. USA 2008, 105, 4088; c) S. A.
Rizvi, S. Liu, Z. L. Chen, C. Skau, M. Pytynia, D. R. Kovar, S. J.
Chmura, S. A. Kozmin, J. Am. Chem. Soc. 2010, 132, 7288.
[11] For reviews on small molecules that affect actin dynamics and
the cytoskeleton, see: a) G. Fenteany, S. Zhu, Curr. Top. Med.
9874 www.angewandte.org
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