M. Mitsuya et al. / Bioorg. Med. Chem. Lett. 19 (2009) 2718–2721
2721
13. Benigni, R.; Giuliani, A.; Franke, R.; Gruska, A. Chem. Rev. 2000, 100, 3697.
14. Kalgutkar, A. S.; Dalvie, D. K.; O’Donnell, J. P.; Taylor, T. J.; Sahakian, D. C. Curr.
Drug Metab. 2002, 3, 379.
4-methyl-1,2,4-triazole by copper-catalyzed reaction produced 5
in 56% yield.
In conclusion, we designed a novel class of 3,6-disubstituted 2-
pyridinecarboxamide GK activators based on the crystal informa-
tion from a GK-allosteric activator complex. Chemical modification
led to the discovery of a series of potent GK activators which elim-
inates the aniline group of the lead compound 1 and serve as a
good source for the development of GK activators suitable for clin-
ical study. Further optimization of this series is ongoing to improve
drug-like properties such as ADME/PK and in vivo efficacy. Detailed
SAR and biological profiles of this novel class will be reported
elsewhere.
15. Crystallographic statistics for the GK-compound 2 complex are as follows:
space group P6522, unit cell 80.1, 80.1, 324.0 Å, resolution 2.52 Å, 21,669
reflections from 439,153 observations give 98.3% completeness with Rmerge of
4.7% and mean I/
r (I) of 71.2. The final model containing 3506 protein, 128
water, 1 salt, 12 glucose and 26 compound atoms has an R-factor of 22.1% (Rfree
using 5% of the data 28.2%). Mean temperature factors for the protein and the
ligand are 30.8 and 28.0 Å2, respectively.
16. Protein and crystals were obtained according to established procedures.24
Crystals were soaked in 0.5 mM compound 2 or compound 4 overnight in
mother liquor containing 5% DMSO. Diffraction data were collected on
beamline BL6B of the Photon Factory in KEK, at 100 K. Data processing and
data reduction were carried out using programs from the HKL2000 (HKL
Research, Inc.) and the CCP4 package.25 Compounds were modeled into the
electron density using AFITT (OpenEye Scientific Software). The protein-
compound complex models were refined using CNX (Accelrys) and REFMAC
5
Acknowledgment
from the CCP4 package. The final structures have been deposited in the Protein
Data Bank with the deposition code 3A0I and 3GOI together with structure
factors and detailed experimental conditions.
We thank Dr. Norikazu Ohtake and Dr. Junichi Sakaki for helpful
discussion.
17. Substituents such as alkyloxy, alkylthio and aryloxy groups at the 3-position
were tested. However, they reduced GK potency.
18. According to the literature,26 EC50 values were measured at 2.5 mM glucose
concentration and are the means of at least two or more independent assays.
Compound 1 was used as an internal control across all assay plates for data
References and notes
validation. The EC50 values of 1 are 0.42 0.09 lM. Maximal GK activation by
compounds 4–14 was in the same range as compound 1 (8.0–8.3-fold over
control levels).
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19. Crystallographic statistics for the GK-compound 4 complex are as follows:
space group P6522, unit cell 79.8, 79.8, 322.0 Å, resolution 2.20 Å, 30,342
reflections from 111,693 observations give 94.5% completeness with Rmerge of
6.4% and mean I/
r (I) of 16.5. The final model containing 3506 protein, 137
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water, 1 salt, 12 glucose and 30 compound atoms has an R-factor of 22.6% (Rfree
using 5% of the data 27.7%). Mean temperature factors for the protein and the
ligand are 36.1 and 24.8 Å2, respectively.
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test. The rats were orally administrated either compound 7 or vehicle alone
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challenge (2 g/kg). Blood glucose concentrations were measured just prior to
and following the glucose challenge (30, 60, 90, 120, 150 and 210 min).
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