Journal of the American Chemical Society
Page 4 of 5
(4) (a) Spokoyny, A. M.; Zou, Y.; Ling, J. J.; Yu, H.; Lin, Y. S.; Pen-
(KD = 151 ± 2 nM), while the biotinylated pDI 12c, which serves
as a reference, falls in the same range as 14c (KD = 66 ± 2 nM).
telute, B. L. J. Am. Chem. Soc. 2013, 135, 5946. (b) Brown, S.; Smith, A.
J. Am. Chem. Soc. 2015, 137, 4034. (c) Vinogradova, E. V.; Zhang, C.;
Spokoyny, A. M.; Pentelute, B. L.; Buchwald, S. L. Nature 2015, 526,
687. (d) Kalhor-Monfared, S.; Jafari, M. R.; Patterson, J. T.; Kitov, P. I.;
Dwyer, J. J.; Nuss, J. M.; Derda, R. Chem. Sci. 2016, 7, 3785.
(5) Lau, Y. H.; de Andrade, P.; Wu, Y.; Spring, D. R. Chem. Soc. Rev.
2015, 44, 91.
(6) (a) Walensky, L. D.; Kung, A. L.; Escher, I.; Malia, T. J.; Barbuto, S.;
Wright, R. D.; Wagner, G.; Verdine, G. L.; Korsmeyer, S. J. Science 2004,
305, 1466. (b) Walensky, L. D.; Pitter, K.; Morash, J.; Oh, K. J.; Barbuto,
S.; Fisher, J.; Smith, E.; Verdine, G. L.; Korsmeyer, S. J. Mol. Cell 2006,
24, 199. (c) Gavathiotis, E.; Reyna, D. E.; Davis, M. L.; Bird, G. H.; Wa-
lensky, L. D. Mol. Cell 2010, 40, 481. (d) Grossmann, T. N.; Yeh, J. T.
H.; Bowman, B. R.; Chu, Q.; Moellering, R. E.; Verdine, G. L. Proc. Natl.
Acad. Sci. U.S.A. 2012, 109, 17942. (e) Chang, Y. S.; Graves, B.; Guer-
lavais, V.; Tovar, C.; Packman, K.; To, K. H.; Olson, K. A.; Kesavan, K.;
Gangurde, P.; Mukherjee, A.; Baker, T.; Darlak, K.; Elkin, C.; Filipovic,
Z.; Qureshi, F. Z.; Cai, H.; Berry, P.; Feyfant, E.; Shi, X. E.; Horstick, J.;
Annis, D. A.; Manning, A. M.; Fotouhi, N.; Nash, H.; Vassilev, L. T.;
Sawyer, T. K. Proc. Natl. Acad. Sci. U. S. A. 2013, 110, E3445. (f) Wa-
lensky, L. D.; Bird, G. H. J. Med. Chem. 2014, 57, 6275.
1
2
3
4
5
6
7
8
In summary, we present the first lysine N-arylation of unpro-
tected peptides and its application to macrocyclization. This
methodology enables the efficient access to a large variety of
macrocyclized scaffolds under mild conditions using numerous
electrophiles. Dimethylformamide was used as the primary reac-
tion solvent to solubilize both the unprotected peptide and elec-
trophile. The use of organic solvent will hinder efforts to modify
proteins in one step and our future efforts are to design soluble
electrophiles for N-arylation of proteins. Since there is much need
for new macrocyclization methodologies that produce shelf and
solution stable constructs, our approach expands this chemical
toolkit and addresses the possible chemical stability issues with
cysteine arylation. Building on these findings, we evolved a
known MDM2 inhibitor into a perfluoroaromatic N-arylated mac-
rocyclic peptide that displayed many desirable characteristics that
may prompt additional cancer studies.
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
ASSOCIATED CONTENT
(7) (a) Blackwell, H. E.; Grubbs, R. H. Angew. Chem., Int. Ed. 1998, 37,
3281. (b) Schafmeister, C. E.; Po, J.; Verdine, G. L. J. Am. Chem. Soc.
2000, 122, 5891. (c) Verdine, G.; Hilinski, G. Methods in Enzymology.
2012, 503, 3.
(8) (a) Felix, A. M.; Heimer, E. P.; Wang, G. T.; Lambros, T. J.; Four-
nier, A. J.; Mowles, T. F.; Maines, S.; Campbell, R. M.; Wegrzynski, B.
B.; Toomer, V.; Fry, D.; Madison, V. S. Int. J. Peptide Protein Res. 1988,
21, 441. (b) Taylor, J. W. Pept. Sci. 2002, 66, 49. (c) de Araujo, A. D.;
Hoang, H. N.; Kok, W. M.; Diness, F.; Gupta, P.; Hill, T. A.; Driver, R.
W.; Price, D. A.; Liras, S.; Fairlie, D. P. Angew. Chem., Int. Ed. 2014, 53,
6965.
(9) (a) Scrima, M.; Le Chevalier-Isaad, A.; Rovero, P.; Papini, A. M.;
Chorev, M.; Ursi, A. M. D. Eur. J. Org. Chem. 2010, 446. (b) Lau, Y.; de
Andrade, P.; Quah, S.-T.; Rossmann, M.; Laraia, L.; Skold, N.; Sum, T.;
Rowling, P.; Joseph, T.; Verma, C.; Hyvonen, M.; Itzhaki, L.; Venkitara-
man, A.; Brown, C.; Lane, D.; Spring, D. Chem. Sci. 2014, 5, 1804.
(10) Jackson, D.; King, D.; Chmielewski, J.; Singh, S.; Schultz, P. J. Am.
Chem. Soc. 1991, 113, 9391.
(11) (a) Muppidi, A.; Doi, K.; Edwardraja, S.; Drake, E.; Gulick, A.;
Wang, H.; Lin, Q. J. Am. Chem. Soc. 2012, 134, 14734. (b) Jo, H.; Mein-
hardt, N.; Wu, Y.; Kulkarni, S.; Hu, Z.; Low, K. E.; Davies, P. L.;
DeGrado, W. F.; Greenbaum, D. C. J. Am. Chem. Soc. 2012, 134, 17704.
(c) Wang, Y.; Chou, D. Angew. Chem., Int. Ed. 2015, 54, 10931. (d)
Assem, N.; Ferreira, D.; Wolan, D.; Dawson, P. Angew. Chem., Int. Ed.
2015, 54, 8665.
(12) (a) S.W.Baertschi,P.J.Jansen, K. M. Alsante in Pharmaceutical Stress
Testing:Predicting Drug Degradation, 2nd. ed. (Eds.: S. W. Baertschi, K.
M. Alsante,R.A.Reed), Informa Healthcare,London, 2011. (b) Hovorka, S.
W.; Schoneich, C. J. Pharm. Sci. 2001, 90, 253. (c) Shen, B. Q.; Xu, K.;
Liu, L.; Raab, H.; Bhakta, S.; Kenrick, M.; Parsons-Reponte, K. L.; Tien,
J.; Yu, S. F.; Mai, E.; Li, D.; Tibbitts, J.; Baudys, J.; Saad, O. M.; Scales,
S. J.; McDonald, P. J.; Hass, P. E.; Eigenbrot, C.; Nguyen, T.; Solis, W.
A.; Fuji, R. N.; Flagella, K. M.; Patel, D.; Spencer, S. D.; Khawli, L. A.;
Ebens, A.; Wong, W. L.; Vandlen, R.; Kaur, S.; Sliwkowski, M. X.;
Scheller, R. H.; Polakis, P.; Junutula, J. R. Nat. Biotechnol. 2012, 30, 184.
(d) Toda, N.; Asano, S.; and Barbas, C. F. Angew. Chem., Int. Ed. 2013,
52, 12592.
(13) Phan, J.; Li, Z.; Kasprzak, A.; Li, B.; Sebti, S.; Guida, W.;
Schonbrunn, E.; Chen, J. J. Biol. Chem. 2010, 285, 2174.
(14) Fahey, D. R.; Ash, C. E. Macromolecules 1991, 24, 4242.
(15) (a) Blotny, G. Tetrahedron 2006, 62, 9507. (b) Shannon, D. A.;
Banerjee, R.; Webster, E. R.; Bak, D. W.; Wang, C.; Weerapana, E. J. Am.
Supporting Information
Full experimental and characterization details. This material is
AUTHOR INFORMATION
Corresponding Author
*blp@mit.edu
Notes
The authors declare no competing financial interests.
ACKNOWLEDGMENT
We thank Prof. Stephen L. Buchwald (S.L.B.) for his encourage-
ment and support. This work was supported by an MIT start-up
fund, the National Institutes of Health (NIH; R01GM110535) and
the Sontag Foundation Distinguished Scientist Award (to B.L.P.).
This work was partially funded by HFSPO (F.T.), and by Bio-
physics grant (P.D.). The authors acknowledge the Biological
Instrument Facility of MIT for providing the Octet BioLayer In-
terferometry System (NIH S10 OD016326), the Koch Institute for
providing the flow cytometer. We thank Dr. Amy Rabideau for
MDM2 protein expression, Colin Fadzen for his help to use FACS
instrument, Chi Zhang and Ethan Evans for technical assistance
and fruitful discussions throughout the course of the work.
REFERENCES
(1) (a) Hili, R.; Yudin, A. K. Nat. Chem. Biol. 2006, 2, 284. (b) Fischer,
C.; Koenig, B. Beilstein J. Org. Chem. 2011, 7, 59. (c) Bariwal, J.; Van
der Eycken, E. V. Chem. Soc. Rev. 2013, 42, 9283.
(2) (a) Cano, R.; Ramon, D. J.; Yus, M. J. Org. Chem. 2011, 76, 654. (b)
Diness, F.; Fairlie, D. P. Angew. Chem., Int. Ed. 2012, 51, 8012.
(3) (a) Yang, B. H.; Buchwald, S. L. J. Organomet. Chem. 1999, 576,
125. (b) Beletskaya, I.; Cheprakov, A. Coord. Chem. Rev. 2004, 248,
2337.
Chem.
Soc.
2014,
136,
3330.
Insert Table of Contents artwork here
ACS Paragon Plus Environment