10.1002/chem.201700874
Chemistry - A European Journal
FULL PAPER
characterized by in vitro acquired resistance to doxorubicin
(DX).
Keywords: tubulysins • anticancer • aza-Michael • in vitro
tests • in vivo tests
In contrast to paclitaxel and in analogy with vinorelbine,
compounds 24e and 24g inhibited tubulin polymerization in
STO, MESOII, LoVo, and LoVo/DX cell lines. A vinorelbine-like
mechanism of action of these tubulysin derivatives was proved
by assessing the capacity of 24e and 24g to antagonize the
paclitaxel tubulin polymerization effect. Moreover, as for
vinorelbine and paclitaxel, mitotic arrest and apoptosis in
cancer cells were detected upon 24e and 24g cell treatments.
These tubulysin derivatives exerted the same effect of the
reference chemotherapeutics, but at significantly lower
concentrations.
In contrast to previously reported studies on natural
tubulysins, these analogues showed effective therapeutic
windows in vivo. In fact, assays in animal models (mice)
of DMPM tumors, evidenced a significant antitumor
activity of 24e at the dose of 0.125 mg/kg (maximum tumor
volume inhibition of 64% and 77% compared to untreated
animals for STO and MESOII, respectively). In STO
xenografts, tumor growth delay induced by i.v.
administration of 24e was less marked than that elicited
by vinorelbine at the dose of 5 mg/kg. It is important to
note that in contrast to vinorelbine (5 mg/kg), 24e (0.125
mg/kg) was able to inhibit also MESOII xenografts. In
addition, the compound was well tolerated with no sign of
general toxicity and a restrained effect (<10%) in terms of
body weight loss.
[1] F. Sasse, H. Steinmetz, J. Heil, G. Höfle, H. Reichenbach, J.
Antibiotics 2000, 53, 879-885.
[2] A. Ullrich, J. Herrmann, R. Muller, U. Kazmaier, Eur. J. Org. Chem.
2009, 6367-6378.
[3] H. Steinmetz, N. Glaser, E. Herdtweck, F. Sasse, H. Reichenbach,
G. Höfle, Angew. Chem. Int. Ed. 2004, 43, 4888-4892.
[4] A. W. Patterson, H. M. Peltier, F. Sasse, J. A. Ellman, Chem. Eur. J.
2007, 13, 9534-9541.
[5] T. Shibue, I. Okamoto, N. Morita, H. Morita, Y. Hirasawa, T. Hosoya,
O. Tamura, Bioorg. Med. Chem. Lett. 2011, 21, 431-434.
[6] G. Kaur, M. Hollingshead, S. Holbeck, V. Schauer-Vukašinović, R.
F. Camalier, A. Dömling, S. Agarwal, Biochem. J. 2006, 396, 235-
242.
[7] R. Balasubramanian, B. Raghavan, A. Begaye, D. L. Sackett, R. A.
Fecik, J. Med. Chem. 2009, 52, 238-240.
[8] O. F. Lamidi, M. Sani, P. Lazzari, M. Zanda, I. N. Fleming, J. Cancer
Res. Clin. Oncol. 2015, 141, 1575-1583.
[9] B. C. Murray, M. T. Peterson, R. A. Fecik, Nat. Prod. Rep. 2015, 32,
654-662.
[10] A. Sandmann, F. Sasse, R. Muller, Chem. Biol. 2004, 11, 1071-
1079.
[11] F. Sasse, D. Menche, Nat. Chem. Biol. 2007, 3, 87-89.
[12] H. M. Peltier, J. P. McMahon, A. W. Patterson, J. A. Ellman, J. Am.
Chem. Soc. 2006, 128, 16018-16019.
[13] O. Pando, S. Dörner, R. Preusentanz, A. Denkert, A. Porzel, W.
Richter, L. Wessjohann, Org. Lett. 2009, 11, 5567-5569.
[14] a) R. Wang, P. Tian, G. Lin, Chin. J. Chem. 2013, 31, 40-48. b) W.
Tao, W. Zhou, Z. Zhou, C.-M. Si, X. Sun, B.-G. Wei, Tetrahedron
2016, 72, 5928-5933.
These results highlight the potential of these tubulysin
derivatives as chemotherapeutics, particularly for treating
currently untreatable tumors, such as diffuse malignant
peritoneal mesotheliomas. In addition, these highly potent
tubulysins may represent very promising payloads in ADCs for
targeted cancer therapy.
[15] M. Sani, G. Fossati, F. Huguenot, M. Zanda, Angew. Chem. Int. Ed.
2007, 46, 3526-3529.
[16] X. D. Yang, C. M. Dong, J. Chen, Y. H. Ding, Q. Liu, X. Y. Ma, Q.
Zhang, Y. Chen, Chem. Asian J. 2013, 8, 1213-1222.
[17] A. Dömling, B. Beck, U. Eichelberger, S. Sakamuri, S. Menon, Q. Z.
Chen, Y. Lu, L. A. Wessjohann, Angew. Chem. Int. Ed. 2006, 45,
7235-7239. Corrigendum: Angew. Chem. Int. Ed. 2007, 46, 2347.
[18] W. Richter, WO 2008/138561 A1, 2008.
[19] O. Pando, S. Stark, A. Denkert, A. Porzel, R. Preusentanz, L. A.
Wessjohann, J. Am. Chem. Soc. 2011, 133, 7692-7695.
[20] S. Rath, J. Liebl, R. Fűrst, A. Ullrich, J. L. Burkhart, U. Kazmaier, J.
Herrmann, R. Műller, M. Gűnther, L. Schreiner, E. Wagner, A. M.
Vollmar, S. Zahler, Br. J. Pharmacol. 2012, 167, 1048-1061.
[21] P. S. Shankar, M. Jagodzinska, L. Malpezzi, P. Lazzari, I. Manca, I.
R. Greig, M. Sani, M. Zanda, Org. Biomol. Chem. 2013, 11, 2273-
2287.
Experimental Section
Chemistry. All the synthetic procedures, compounds
characterizations and copies of 1H, 13C, 19F NMR spectra and
MS analyses are included in the Supporting Information.
Biology. Procedures and materials on in vitro assays, cell lines
and antiproliferative activity assays, tubulin polymerization
assays, in vivo studies, as well as details on methodologies
[22] P. S. Shankar, S. Bigotti, P. Lazzari, I. Manca, M. Spiga, M. Sani, M.
Zanda, Tetrahedron Lett. 2013, 54, 6137-6141.
[23] Z. Wang, P. A. McPherson, B. S. Raccor, R. Balachandran, G. Zhu,
B. W. Day, A. Vogt, P. Wipf, Chem. Biol. Drug Des. 2007, 70, 75-86.
[24] T. Shibue, I. Okamoto, N. Morita, H. Morita, Y. Hirasawa, T. Hosoya,
O. Tamura, Bioorg. Med. Chem. Lett. 2011, 21, 431-434.
[25] X. Yang, C. Dong, J. Chen, Q. Liu, B. Han, Q. Zhang, Y. Chen,
Tetrahedron Lett. 2013, 54, 2986-2988.
(western
immunoblotting
flow
cytometer
analyses,
fluorescence microscopy) are described in the Supporting
information.
[26] S. P. Shankar, M. Sani, M F. R. Saunders, H. M. Wallace, M. Zanda,
Synlett 2011, 1673-1676.
Acknowledgements
[27] T. Schluep, P. Gunawan, L. Ma, G. S. Jensen, J. Duringer, S.
Hinton, W. Richter, J. Hwang, Clin. Cancer Res. 2009, 15, 181-189.
[28] C. P. Leamon, J. A. Reddy, M. Vetzel, R. Dorton, E. Westrick, N.
Parker, Y. Wang, I. Vlahov, Cancer Res. 2008, 68, 9839-9844.
We thank Regione Autonoma della Sardegna RAS (Italy) for
economic support by covering in part the costs of this research.
I.U. acknowledges RAS for his fellowship (for grant numbers
see the Supporting Information).
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