Journal of the American Chemical Society
COMMUNICATION
’ ACKNOWLEDGMENT
We are grateful for financial support of this research from the
National Cancer Institute (1R15CA152914-01) and UWM start-up
funds (X.P.). This research was supported in part by the National
Cancer Institute (-CA136411) Lymphoma SPORE (V.G.).
’ REFERENCES
(1) (a) Noll, D. M.; Mason, T. M.; Miller, P. S. Chem. Rev. 2006,
106, 277. (b) Dronkert, M. L. G.; Kanaar, R. Mutat. Res. 2001, 486, 217.
(2) (a) Hong, I. S.; Greenberg, M. M. J. Am. Chem. Soc. 2005,
127, 10510. (b) Hong, I. S.; Ding, H.; Greenberg, M. M. J. Am. Chem. Soc.
2006, 128, 485. (c) Peng, X.; Hong, I. S.; Li, H.; Seidman, M. M.;
Greenberg, M. M. J. Am. Chem. Soc. 2008, 130, 10299.
(3) (a) Veldhuyzen, W. F.; Pande, P.; Rokita, S. E. J. Am. Chem. Soc.
2003, 125, 14005. (b) Richter, S. N.; Maggi, S.; Mels, S. C.; Palumbo, M.;
Freccero, M. J. Am. Chem. Soc. 2004, 126, 13973. (c) Wang, H.; Rokita,
S. E. Angew. Chem., Int. Ed. 2010, 49, 5957.
(4) (a) Wang, P.; Liu, R.; Wu, X.; Ma, H.; Cao, X.; Zhou, P.; Zhang,
J.; Weng, X.; Zhang, X.; Qi, J.; Zhou, X.; Weng, L. J. Am. Chem. Soc. 2003,
125, 1116. (b) Weng, X.; Ren, L.; Weng, L.; Huang, J.; Zhu, S.; Zhou, X.;
Weng, L. Angew. Chem., Int. Ed. 2007, 46, 8020.
(5) (a)Shawn, M. H.;Marquis, J. C.;Zayas, B.;Wishnok, J. S.;Liberman,
R. G.; Skipper, P. L.; Tannenbaum, S. R.; Essigmann, J. M.; Croy, R. G. Mol.
Cancer Ther. 2006, 5, 977. (b) Rink, S. M.; Yarema, K. J.; Solomon, M. S.;
Paige, L. A.; Mitra Tadayoni-Rebek, B.; Essigmann, J. M.; Croy, R. G. Proc.
Natl. Acad. Sci. U.S.A. 1996, 93, 15063. (c) Kim, E.; Rye, P. T.; Essigmann,
J. M.; Croy, R. G. J. Inorg. Biochem. 2009, 103, 256.
Figure 4. Effect of compounds 1ꢀ3 on cancer cells and normal
lymphocytes: (A) four human cancer cells (SR, NCI-H460, CAKI-1,
and SN12C)) were incubated with 10 μM of compounds 1ꢀ3 for 48 h
(gray bar, 3; black bar, 1; lined bar, 2); (B) normal lymphocytes obtained
from healthy donors (n = 3) were incubated with 10 μM of 1 and 2 for 48
h. Time matched control samples are set up concurrently (gray bar,
control; black bar, compound 1; lined bar, compound 2).
boronic acid group in 2 inhibits the oxidative ability of H2O2. It is
precedented that the reaction between arylboronic acid and
hydrogen peroxide was pH-dependent.11,17
Having established that the prodrugs 1 and 2 could be
effectively passivated and activated by H2O2, the ability of these
compounds to inhibit cancer cell growth was evaluated. Both
compounds inhibited various types of cancer cells at 10 μM.
They showed about 90% inhibition toward SR cells (Leukemia
cell), 85% inhibition toward NCI-H460 (Nonsmall Cell Lung
Cancer cells), and 66% inhibition toward CAKI-1, and 57%
toward SN12C (Renal Cancer cells) (Figure 4A).18 However,
compound 3 is less toxic to these cells. The toxicity of 1 and 2 is
highly likely caused by the release of nitrogen mustard after
tumor-specific activation. To determine the selectivity, we eval-
uated the toxicity of 1 and 2 toward noncancer cells. Normal
lymphocytes obtained from three healthy donors were incubated
without or with 10 μM of compounds 1 and 2; untreated samples
were used as time-matched controls. In all the 3 samples studied,
compared to time-matched controls, there was no increase in
apoptosis observed at 24ꢀ72 h (Figure 4B and SI, Figure S11).
In conclusion, two prodrugs of nitrogen mustard coupled with an
arylboronate or boronic acid demonstrated an effective way to mask
the cytotoxicity of cancer chemotherapeutic agents and selectively
release them in the presence of H2O2. The activity and selectivity
were measured by cross-linking or alkylation of DNA as well as by
evaluating their ability to inhibit cancer cell growth and toxicity
toward normal cells. To the best of our knowledge, we are the first to
report anticancer prodrugs (1 and 2) that can be activated by ROS
to release DNA cross-linking agents. Such compounds are nontoxic
but are highly likely to undergo tumor-specific activation to generate
toxic species in cancer cell. They offer novel ways to improve the
therapeutic effectiveness and selectivity of current anticancer agents.
(6) Szatrowski, T. P.; Nathan, C. F. Cancer Res. 1991, 51, 794.
(7) Toyokuni, S.; Okamoto, K.; Yodoi, J.; Hiai, H. FEBS Lett. 1995,
358, 1.
(8) Kawanishi, S.; Hiraku, Y.; Pinlaor, S.; Ma, N. Biol. Chem. 2006, 365.
(9) (a) Pelicano, H.; Carney, D.; Huang, P. Drug Resist. Updates
2004, 7, 97. (b) Trachootham, D.; Alexandre, J.; Huang, P. Nat. Rev.
2009, 8, 579. (c) Lopez-Lazaro, M. Cancer Lett. 2007, 252, 1.
(10) (a) Zieba, M.; Suwalski, M.; Kwiatkowska, S.; Piasecka, G.;
Grzelewska-Rzymowska, I.; Stolarek, R.; Nowak, D. Respir. Med. 2000,
94, 800. (b) Lim, S. D.; Sun, C.; Lambeth, J. D.; Marshall, F.; Amin, M.;
Chung, L.; Petros, J. A.; Arnold, R. S. Prostate 2005, 62, 200.
(11) Kuivila, H. G.; Armour, A. G. J. Am. Chem. Soc. 1957, 79, 5659.
(12) (a) Miller, E. W.; Albers, A. E.; Pralle, A.; Isacoff, E. Y.; Chang,
C. J. J. Am. Chem. Soc. 2005, 127, 16652. (b) Dickinson, B. C.; Chang,
C. J. A. J. Am. Chem. Soc. 2008, 130, 9638. (c) Miller, E. W.; Tulyathan,
O.; Isacoff, E. Y.; Chang, C. J. Nat. Chem. Biol. 2007, 3, 263. (d) Lo, L. C.;
Chu, C. Y. Chem. Commun. 2003, 2728. (e) Srikun, D.; Miller, E. W.;
Domaille, D. W.; Chang, C. J. A. J. Am. Chem. Soc. 2008, 130, 4596.
(13) Yang, W. Q.; Gao, X.; Wang, B. H. In Boronic Acids; Hall, D. G.,
Ed.; Wiley-VCH: Weinheim, 2005; pp 481ꢀ512.
(14) (a) Brookes, P.; Lawley, P. D. Biochem. J. 1961, 80, 496. (b)
Rink, S. M.; Solomon, M. S.; Taylor, M. J.; Rajur, S. B.; McLaughlin,
L. W.; Hopkins, P. B. J. Am. Chem. Soc. 1993, 115, 2551. (c) Dong, Q.;
Barsky, D.; Colvin, M. E.; Melius, C. F.; Ludeman, S. M.; Moravek, J. F.;
Colvin, O. M.; Bigner, D. D.; Modrich, P. Proc. Natl. Acad. Sci. U.S.A.
1995, 92, 12170.
’ ASSOCIATED CONTENT
(15) Haraguchi, K.; Delaney, M. O.; Wiederholt, C. J.; Sambandam,
A.; Hantosi, Z.; Greenberg, M. M. J. Am. Chem. Soc. 2002, 124, 3263.
(16) Weinert, E. E.; Dondi, R.; Colloredo-Melz, S.; Frankenfield,
K. N.; Mitchell, C. H.; Freccero, M.; Rokita, S. E. J. Am. Chem. Soc. 2006,
128, 11940.
S
Supporting Information. Experimental procedures for all
b
reaction and analyses, preparation and characterization of 1ꢀ7,
bioactive evaluation, autoradiograms of Fe EDTA, and piperidine
3
treatment of cross-linked products and reacted single-stranded
DNA. This material is available free of charge via the Internet at
(17) Kuivila, H. G. J. Am. Chem. Soc. 1954, 76, 870.
(18) Data were obtained from Developmental Therapeutics Pro-
gram at the National Cancer Institute (NCI-60 DTP Human Tumor
Cell Line Screen).
’ AUTHOR INFORMATION
Corresponding Author
19281
dx.doi.org/10.1021/ja2073824 |J. Am. Chem. Soc. 2011, 133, 19278–19281