X.-G. Bai et al. / Bioorg. Med. Chem. Lett. 22 (2012) 6947–6951
6951
The above results suggested that acetyl and benzamido groups
be 4- to 28-fold more potent than that of (R)-roscovitine (IC50
:
were the optimal substituents at the N1- and C3-positions in this
study, respectively. In order to define more clearly the effect the
substituent at the N5-position on the activity, the benzoyl moiety
at the N5-position of the most active compound 11a was also re-
placed with other acyl and sulfonyl groups (Table 2). When directly
compared with 11a, these derivatives 12a–k showed remarkably
decreased activity, although some of them (12a, f, g, k) were more
active than (R)-roscovitine against HCT-116.
9.62-16.38 M) against the six human cancer cell lines. In addition,
l
11a has promising inhibitory activity against both CDK5/p25 and
GSK3b. Docking results found that 11a share the similar interaction
mode with CDK5 as (R)-roscovitine, 11a also formed hydrogen
bond and
p–p interactions with GSK3b. Further pharmacological
studies of 11a are currently in progress and will be reported in
due course.
Compounds 11a and 11b were further evaluated for their
in vitro anticancer activity against other five human cancer cell
lines including HT-29, MCF7, HepG2, A549 and HT-1080 as well
as one normal human liver L02 cell line. The data in Table 3 indi-
cated that 11a and 11b have more remarkable anticancer activity
against the five human cancer cell lines and human colon carci-
Acknowledgments
We are grateful to the financial support from the National S&T
Major Special Project on Major New Drug Innovation
(2012ZX093101002-001-017) and the National Natural Science
Foundation of China (No. 81072577 and 81102376).
noma HCT-116 cell line (Table 1) (IC50: 0.58–2.13
lM) compared
with (R)-roscovitine (IC50: 9.62–16.38 M). On the other hand,
compound 11a showed promising anticancer activity on the six
l
Supplementary data
human cancer cell lines than normal human liver L02 cell line
Supplementary data associated with this article can be found, in
(IC50: 2.22
Finally, 11a was assessed for its activity against a panel of 12 ki-
nases at 10 M concentration. The data in Table 4 indicated that
lM), which is consistent with (R)-roscovitine.
l
11a has much better activity against CDK5/p25 and GSK3b (72%
and 67% of inhibition rates, respectively) than the other ten kinases
(3–33% of inhibition rates).
References and notes
1. Anwara, H. F.; Elnagdic, M. H. Arkivoc 2009, 1, 198.
2. Boss, D.; Witteveen, P.; Van Der Sar, J.; Lolkema, M.; Voest, E.; Stockman, P.;
Ataman, O.; Wilson, D.; Das, S.; Schellens, J. Ann. Oncol. 2011, 22, 431.
3. Tyler, R. K.; Shpiro, N.; Marquez, R.; Eyers, P. A. Cell Cycle 2007, 6, 2846.
4. Cohen, R. B.; Jones, S. F.; Aggarwal, C.; von Mehren, M.; Cheng, J.; Spigel, D. R.;
Greco, F. A.; Mariani, M.; Rocchetti, M.; Ceruti, R. Clin. Cancer Res. 2009, 15,
6694.
5. Pevarello, P.; Brasca, M. G.; Orsini, P.; Traquandi, G.; Longo, A.; Nesi, M.; Orzi, F.;
Piutti, C.; Sansonna, P.; Varasi, M. J. Med. Chem. 2005, 48, 2944.
6. Fancelli, D.; Berta, D.; Bindi, S.; Cameron, A.; Cappella, P.; Carpinelli, P.; Catana,
C.; Forte, B.; Giordano, P.; Giorgini, M. L. J. Med. Chem. 2005, 48, 3080.
7. Fancelli, D.; Moll, J.; Varasi, M.; Bravo, R.; Artico, R.; Berta, D.; Bindi, S.;
Cameron, A.; Candiani, I.; Cappella, P. J. Med. Chem. 2006, 49, 7247.
8. Pevarello, P.; Fancelli, D.; Vulpetti, A.; Amici, R.; Villa, M.; Pittala, V.; Vianello,
P.; Cameron, A.; Ciomei, M.; Mercurio, C.; Bischoff, J. R.; Roletto, F.; Varasi, M.;
Brasca, M. G. Bioorg. Med. Chem. Lett. 2006, 1084, 16.
9. Shi, J.; Xu, G.; Zhu, W.; Ye, H.; Yang, S.; Luo, Y.; Han, J.; Yang, J.; Li, R.; Wei, Y.
Bioorg. Med. Chem. Lett. 2010, 20, 4273.
10. Liao, J. J. J. Med. Chem. 2007, 50, 409.
11. Jain, P.; Flaherty, P. T.; Yi, S.; Chopra, I.; Bleasdell, G.; Lipay, J.; Ferandin, Y.;
Meijer, L.; Madura, J. D. Bioorg. Med. Chem. 2011, 19, 359.
12. Misra, R. N.; Xiao, H.; Rawlins, D. B.; Shan, W.; Kellar, K. A.; Mulheron, J. G.;
Sack, J. S.; Tokarski, J. S.; Kimball, S. D.; Webster, K. R. Bioorg. Med. Chem. Lett.
2003, 13, 2405.
To further understand the binding mode of compound 11a with
CDK5 or GSK3b, molecular docking was performed through CDOC-
KER module in Discover studio (Fig. 3). Compound 11a packs nicely
into the ATP binding pockets of both CDK5 and GSK3b (Fig. 3a and
3c). The superimposition of the binding modes of 11a and (R)-ros-
covitine in the ATP binding pocket of CDK5 was shown as Figure 3b.
The acetyl of 11a and the isopropyl of (R)-roscovitine form similar
hydorphobic interactions with the narrow hydrophobic pocket
around residue Phe80. Furthermore the C-3-benzamido and N-5-
benzoyl moieties of 11a shows similar orietation as the N-6-benzyl
and 1-ethyl-2-hydroxyethylamino moieties of (R)-roscovitine,
respectively, and the 1-acyl-3-aminopyrazole moiety of 11a also
forms two critical H-bonds with residue CYS83. A third H-bond
formed between N-5-benzoyl moiety of 11a and residue ASN144.
Shown as Figure 3d, there is only one H-bond formed between
C-3 amino group and the oxygen of ASP200 residue of GSK3b.
Meanwhile, C-3-benzamido and N-5-benzoyl moieties of 11a form
two
tively. The
p
–
p
interactions with residues LYS183 and ARG141, respec-
interaction formed between the phenyl moiety of
13. Johnson, L. N.; De Moliner, E.; Brown, N. R.; Song, H.; Barford, D.; Endicott, J. A.;
Noble, M. E. M. Pharmacol. Therapeut. 2002, 93, 113.
p–p
14. Sanchez-Martinez, C.; Shih, C.; Zhu, G.; Li, T.; Brooks, H. B.; Patel, B. K. R.;
Schultz, R. M.; DeHahn, T. B.; Spencer, C. D.; Watkins, S. A. Bioorg. Med. Chem.
Lett. 2003, 13, 3841.
15. Wyatt, P. G.; Woodhead, A. J.; Berdini, V.; Boulstridge, J. A.; Carr, M. G.; Cross, D.
M.; Davis, D. J.; Devine, L. A.; Early, T. R.; Feltell, R. E. J. Med. Chem. 2008, 51,
4986.
16. Krasavin, M.; Konstantinov, I. O. Lett. Org. Chem. 2008, 5, 594.
17. Skehan, P.; Storeng, R.; Scudiero, D.; Monks, A.; McMahon, J.; Vistica, D.;
Warren, J. T.; Bokesch, H.; Kenney, S.; Boyd, M. R. J. Natl. Cancer I. 1990, 82,
1107.
18. Tavares, F. X.; Boucheron, J. A.; Dickerson, S. H.; Griffin, R. J.; Preugschat, F.;
Thomson, S. A.; Wang, T. Y.; Zhou, H. Q. J. Med. Chem. 2004, 47, 4716.
19. Kuo, G. H.; Prouty, C.; DeAngelis, A.; Shen, L.; O’Neill, D. J.; Shah, C.; Connolly, P.
J.; Murray, W. V.; Conway, B. R.; Cheung, P. J. Med. Chem. 2003, 46, 4021.
11a N-5-benzoyl group and the C=NH of residue ARG141, which
is consistent to previous modeling studies that the ARG141 residue
(GSK3b) appears to be unique to GSK3b and the interactions with it
are benefitial to the selectivity of GSK3b inhibitors.18,19
In summary, a series of novel 1-acyl-3-amino-1,4,5,6-tetrahy-
dropyrrolo[3,4-c] pyrazole derivatives was designed, synthesized
and characterized by 1H NMR, 13C NMR, MS and HRMS. These
derivatives and three precursors were evaluated for their in vitro
anticancer activity. The results showed that all the tested com-
pounds have generally considerable activity against HCT-116.
The activity of compound 11a (IC50: 0.58–2.13 lM) was found to