Page 9 of 9
Organic & Biomolecular Chemistry
Please do not adjust margins
Journal Name
ARTICLE
4
5
6
(Becton Dickinson, Franklin Lakes, NJ, USA). The ModFit
software (Verity Software House, Topsham, ME, USA) and
CellQuest (Becton Dickinson) software were used to analyze
the data.
Zhan, Epigenomics, 2015, 7, 1379–139D6.OI: 10.1039/C6OB01483F
D. M. Schmidt, and D. G.McCafferty, Biochemistry, 2007, 46,
4408–4416.
M. Yang, J. C. Culhane, L. M. Szewczuk, P. Jalili, H. L. Ball, M.
Machius, P. A. Cole, and H. Yu, Biochemistry, 2007, 46, 8058–
8065.
S. Mimasu, N. Umezawa, S. Sato, T. Higuchi, T. Umehara, and
S. Yokoyama, Biochemistry, 2010, 49, 6494–6503.
Y. Liang, D. Quenelle, J. L. Vogel, C. Mascaro, M. A. Ortega,
and T. M. A. Kristie, mBio., 2013, 4, e00558.
M. A. Ortega, L. J. Castro-Palomino, and M. C. T. Fyfe, Patent
application WO 2011035941, 2011.
Western Blot Analysis. Cells were incubated with the indicated
concentrations of M1310 for 24 h. After washing with PBS, the
cells were lysed in lysis buffer containing 50 mM Tris-HCl
[pH8.0], 150 mM NaCl, 1% NP-40, 0.5% deoxycholic acid, 0.1%
SDS, 1 mM EDTA, 1 mM DTT, 0.5 mM PMSF, 2 mg/mL aprotinin,
2 mg/mL leupeptin, and phosphatase inhibitor cocktail (1/100
v/v, 07575-51, Nacalai Tesque, Kyoto, Japan). The lysate was
sonicated and centrifuged, and the supernatant was subjected
to SDS-PAGE. Rabbit monoclonal anti-histone H3K4me3
(#9751), histone H3K4me2 (#9725), histone H3K4me1 (#5326),
histone H3 (#4499, Cell Signaling Technology, Danvers, MA,
USA), and mouse monoclonal anti-β-actin (A5441, Sigma, St.
Louis, MO, USA) antibodies were used as the primary
antibodies. The blots were incubated with the appropriate
HRP-conjugated secondary antibody (GE Healthcare,
Piscataway, NJ, USA), and signals were detected with Chemi-
Lumi One L (Nacalai Tesque) or Immobilon Western (Millipore,
Bedford, MA, USA).
7
8
9
10 R. Neelamegam, E. L. Ricq, M. Malvaez, D. Patnaik, S. Norton,
S. M. Carlin, I. T. Hill, M. A. Wood, S. J. Haggarty, and J.
Hooker, ACS Chem. Neurosci., 2012, 3, 120–128.
11 M. Yang, J. C. Culhane, L. M. Szewczuk, C. B. Gocke, C. A.
Brautigam, D. R. Tomchick, M. Machius, P. A. Cole, and H. Yu,
Nat. Struct. Mol. Biol., 2007, 14, 535–539.
12 P. Vianello, O. A. Botrugno, A. Cappa, R. Dal Zuffo, P.
Dessanti, A. Mai, B. Marrocco, A. Mattevi, G. Meroni, S.
Minucci, G. Stazi, F. Thaler, P. Trifiró, S. Valente, M.
Villa, M. Varasi, and C. Mercurio, J. Med. Chem., 2016, 59,
1501.
13 R. Ueda, T. Suzuki, K. Mino, H. Tsumoto, H. Nakagawa, M.
Hasegawa, R. Sasaki, T. Mizukami, and N. Miyata, J. Am.
Chem. Soc., 2009, 131, 17536–17537.
Anti-tumor study in xenograft model. Five week-old male
BALB/c nu/nu mice were obtained from Shimizu Laboratory
Supplies Co., Ltd. (Kyoto, Japan). A total of 4.1 x 106 A549 cells
with Matrigel Basement Membrane Matrix (Corning, Corning,
NY, USA) was subcutaneously inoculated into the left flank of
the mice. When the mean tumor volume reached 100 mm3,
the mice were randomized into two groups, and M1310 solved
in PBS containing 15% NIKKOL HCO-60 (PHARM) or vehicle was
intravenously injected thrice a week for 6 weeks. NIKKOL
HCO-60 (PHARM) was provided from Nikko Chemicals Co., Ltd.
(Tokyo, Japan). Tumor volume was measured once a week
with a caliper and calculated by the following formula: 1/2 x
(length x width2). Body weight was also measured once a
week. All experiments and procedures were performed in
accordance with the Institutional Care Use Committee
guidelines.
14 D. Ogasawara, T. Suzuki, K. Mino, R. Ueda, M. N. Khan, T.
Matsubara, K. Koseki, M. Hasegawa, R. Sasaki, H. Nakagawa,
T. Mizukami, and N. Miyata, Bioorg. Med. Chem., 2011, 19,
3702–3708.
15 D. Ogasawara, Y. Itoh, H. Tsumoto, T. Kakizawa, K. Mino, K.
Fukuhara, H. Nakagawa, M. Hasegawa, R. Sasaki, T.
Mizukami, N. Miyata, and T. Suzuki, Angew. Chem. Int. Ed.,
2013, 52, 8620–8624.
16 D. M. Gooden, D. M. Schmidt, J. A. Pollock, A. M. Kabadi, and
D. G. McCafferty, Bioorg. Med. Chem. Lett., 2008, 18, 3047–
3051.
17 H. Benelkebir, C. Hodgkinson, P. J. Duriez, A. L. Hayden, R. A.
Bulleid, S. J. Crabb, G. Packham, and A. Ganesan, Bioorg.
Med. Chem., 2011, 19, 3709–3716.
18 S. Miyamura, M. Araki, T. Suzuki, J. Yamaguchi, and K.
Itami, Angew. Chem. Int. Ed., 2015, 54, 846–851.
19 Although the precise mechanism remains unclear,
epimerization occurred at the nitrogen-bound carbon atom
under N2 atmosphere. See details in ref. 17.
20 During removal of pivaloyl group and thereafter, the
epimerization also occurred to afford trans-ACPA derivatives.
21 T. Kan, and Fukuyama, Chem. Commun., 2004, 353–359.
22 D. M. Schmidt, and D. G. McCafferty, Biochemistry, 2007, 46,
4408–4416.
23 G. W. Humprey, Y. Wang, V. R. Russanova, T. Hirai, J. Qin, Y.
Nakatani, and B. H. Howard, J. Biol. Chem., 2001, 276, 6817–
6824.
24 T. Lv, D. Yuan, X.Miao, Y. Lv, P. Zhan, X. Shen, Y. Song, PLoS
ONE 2012, 7, e35065.
Statistical analysis. Statistical analysis was performed using
Dunnett’s test. P<0.05 was considered significant.
Acknowledgements
We thank Mie Morita for technical support. This work was
supported by KAKENHI (16H01140 and 16H04148 to J.Y.) from
MEXT, and JST CREST program (T.S.). ITbM is supported by the
World Premier Interntional Research Center (WPI) Initiative,
Japan.
25 S. Hayami, J. D. Kelly, H. S. Cho, M. Yoshimatsu, M. Unoki, T.
Tsunoda, H. I. Field, D. E. Neal, H. Yamaue, B. A. Ponder, Y.
Nakamura, and Hamamoto, R. Int. J. Cancer, 2011, 128, 574–
586.
26 C. T. Foster, O. M. Dovey, L. Lezina, J. L. Luo, T. W. Gant, N.
Barlev, A. Bradley, and S. M. Cowley, Mol. Cell Biol., 2010, 30,
4851–4863.
27 A. Adamo, B. Sesé, S. Boue, J. Castaño, I. Paramonov, and M.
J. Barrero, Nat. Cell Biol., 2011, 13, 652–659.
28 M. N. A. Khan, T. Suzuki, and N. Miyata, Med. Res. Rev., 2013,
33, 873–910.
Notes and references
1
Y. Shi, F. Lan, C. Matson, P. Mulligan, J. R. Whetstine, P. A.
Cole, R. A. Casero, and Y. Shi, Cell, 2004, 119, 941–953.
M. Yang, C. B. Gocke, X. Luo, D. Borek, D. R. Tomchick, M.
Machius, Z. Otwinowski, and H. Yu, Mol. Cell, 2006, 23, 377–
387.
2
3
P. Stavropoulos, G. Blobel, and A. Hoelz, Nat. Struct. Mol.
Biol., 2006, 13, 626–632.
This journal is © The Royal Society of Chemistry 20xx
J. Name., 2013, 00, 1-3 | 9
Please do not adjust margins