ACS Medicinal Chemistry Letters
Letter
Synthetic schemes, procedure, and analytical data for 15
and 19; analytical data of 1−14, 16−18, and 20−21;
general animal study protocols; high-throughput sol-
ubility assay protocols (PDF)
AUTHOR INFORMATION
Corresponding Author
■
Notes
The authors declare no competing financial interest.
ABBREVIATIONS
■
ADMET, absorption, distribution, metabolism, excretion,
toxicity; CYP, cytochromes P450; PK/PD, pharmacokinetic/
pharmacodynamics; GPCR, G-protein coupled receptor
Figure 3. PK−PD relationship of 19 in the mouse MMTV-Wnt1
tumor model (n = 3 per time point).
Changes in tumor volume for each of the treated (T) and
control (C) groups were measured, and the T/C ratio was
calculated to express the level of efficacy. As shown in Figure 4,
REFERENCES
■
(1) Barker, N.; Clevers, H. Mining the Wnt pathway for cancer
therapeutics. Nat. Rev. Drug Discovery 2006, 5, 997−1014.
(2) Moon, R. T.; Kohn, A. D.; De Ferrari, G. V.; Kaykas, A. WNT
and beta-catenin signalling: diseases and therapies. Nat. Rev. Genet.
2004, 5, 691−701.
(3) Turashvili, G.; Bouchal, J.; Burkadze, G.; Kolar, Z. Wnt signaling
pathway in mammary gland development and carcinogenesis.
Pathobiology 2006, 73, 213−223.
(4) Braun, K. M. Cutaneous cancer stem cells: beta-catenin strikes
again. Cell Stem Cell 2008, 2, 406−408.
(5) Polakis, P. Wnt signaling in cancer. Cold Spring Harbor Perspect.
Biol. 2012, 4 (5), pii:a00805210.1101/cshperspect.a008052.
(6) Nusse, R.; Varmus, H. Three decades of Wnts: A personal
perspective on how a scientific field developed. EMBO J. 2012, 31,
2670−2684.
(7) Seshagiri, S.; Stawiski, E. W.; Durinck, S.; Modrusan, Z.; Storm,
E. E.; Conboy, C. B.; Chaudhuri, S.; Guan, Y.; Janakiraman, V.; Jaiswal,
B. S.; Guillory, J.; Ha, C.; Dijkgraaf, G. J.; Stinson, J.; Gnad, F.;
Huntley, M. A.; Degenhardt, J. D.; Haverty, P. M.; Bourgon, R.; Wang,
W.; Koeppen, H.; Gentleman, R.; Starr, T. K.; Zhang, Z.; Largaespada,
D. A.; Wu, T. D.; de Sauvage, F. J. Recurrent R-spondin fusions in
colon cancer. Nature 2012, 488, 660−664.
(8) Hao, H. X.; Xie, Y.; Zhang, Y.; Charlat, O.; Oster, E.; Avello, M.;
Lei, H.; Mickanin, C.; Liu, D.; Ruffner, H.; Mao, X.; Ma, Q.; Zamponi,
R.; Bouwmeester, T.; Finan, P. M.; Kirschner, M. W.; Porter, J. A.;
Serluca, F. C.; Cong, F. ZNRF3 promotes Wnt receptor turnover in an
R-spondin-sensitive manner. Nature 2012, 485, 195−200.
(9) Jiang, X.; Hao, H. X.; Growney, J. D.; Woolfenden, S.; Bottiglio,
C.; Ng, N.; Lu, B.; Hsieh, M. H.; Bagdasarian, L.; Meyer, R.; Smith, T.
R.; Avello, M.; Charlat, O.; Xie, Y.; Porter, J. A.; Pan, S.; Liu, J.;
McLaughlin, M. E.; Cong, F. Inactivating mutations of RNF43 confer
Wnt dependency in pancreatic ductal adenocarcinoma. Proc. Natl.
Acad. Sci. U. S. A. 2013, 110, 12649−54.
(10) Giannakis, M.; Hodis, E.; Jasmine, M. X.; Yamauchi, M.;
Rosenbluh, J.; Cibulskis, K.; Saksena, G.; Lawrence, M. S.; Qian, Z. R.;
Nishihara, R.; Van Allen, E. M.; Hahn, W. C.; Gabriel, S. B.; Lander, E.
S.; Getz, G.; Ogino, S.; Fuchs, C. S.; Garraway, L. A. RNF43 is
frequently mutated in colorectal and endometrial cancers. Nat. Genet.
2014, 46, 1264−6.
(11) Gurney, A.; Axelrod, F.; Bond, C. J.; Cain, J.; Chartier, C.;
Donigan, L.; Fischer, M.; Chaudhari, A.; Ji, M.; Kapoun, A. M.; Lam,
A.; Lazetic, S.; Ma, S.; Mitra, S.; Park, I. K.; Pickell, K.; Sato, A.; Satyal,
S.; Stroud, M.; Tran, H.; Yen, W. C.; Lewicki, J.; Hoey, T. Wnt
pathway inhibition via the targeting of Frizzled receptors results in
decreased growth and tumorigenicity of human tumors. Proc. Natl.
Acad. Sci. U. S. A. 2012, 109, 11717−11722.
Figure 4. In vivo antitumor efficacy of 19 in the mouse MMTV-Wnt1
tumor model (n = 8 per group).
compound 19 showed very robust dose-related antitumor
efficacy. Significant tumor growth inhibition (T/C 15%, p <
0.0001) was observed at a dose of 0.3 mg/kg, while tumor
regression was achieved at 1 and 3 mg/kg (T/C −74% and
−84%, respectively, p < 0.0001). The compound was well
tolerated with no significant body weight loss in all treated
groups (data not shown).
In summary, a novel chemical series was developed from
GNF-1331, a screening hit identified by a Wnt coculture
reporter gene assay, as Porcupine inhibitor to block Wnt ligand
secretion and subsequently Wnt signaling activity. Compounds
with exquisite potency and specificity, excellent pharmacoki-
netics, and desirable drug-like properties were identified.
Treatment with 19, GNF-6231, in MMTV-Wnt1 subcuta-
neously implanted tumor bearing mice led to pronounced
inhibition of the Wnt signaling activity as measured by the
reduction of Wnt target gene Axin2 expression. This compound
exhibited robust antitumor efficacy in the same model upon
repeat dosing treatment. Porcupine inhibitors may represent a
new therapeutic approach for diseases with aberrant Wnt
signaling activities.
ASSOCIATED CONTENT
* Supporting Information
The Supporting Information is available free of charge on the
■
S
(12) Ettenberg, S. A.; Charlat, O.; Daley, M. P.; Liu, S.; Vincent, K. J.;
Stuart, D. D.; Schuller, A. G.; Yuan, J.; Ospina, B.; Green, J.; Yu, Q.;
Walsh, R.; Li, S.; Schmitz, R.; Heine, H.; Bilic, S.; Ostrom, L.; Mosher,
R.; Hartlepp, K. F.; Zhu, Z.; Fawell, S.; Yao, Y. M.; Stover, D.; Finan,
D
ACS Med. Chem. Lett. XXXX, XXX, XXX−XXX