Article
Journal of Medicinal Chemistry, 2010, Vol. 53, No. 1 153
Mechanism In ATP Channel Opener-Mediated Protection
Against Streptozotocin-Induced Suppression of Rat Pancreatic
Islet Function. Biochem. Pharmacol. 2008, 76, 1748–1756.
(
m, 1H, 6-H), 10.43 (s, 1H, NH). Anal. (C10
H, N, S.
Ionization Constants. The pK
H
12ClN
3
O
2
S) C,
K
a
values of the compounds were
(
(
7) Uhde, I.; Toman, A.; Gross, I.; Schwanstecher, C.; Schwanstecher,
M. Identification of the Potassium Channel Opener site on Sulfo-
nylurea Receptors. J. Biol. Chem. 1999, 274, 28079–28082.
determined by means of U.V. spectrophotometry using a Perkin-
Elmer UV/vis 554 spectrophotometer at 25 °C. UV spectra of
compounds were taken in different aqueous buffers of pH rank-
ing from 5 to 12. The pK
Debye-H u€ ckel equation at the wavelength giving the maximum
absorbance of the ionized form.
Biological Assays. Measurements of Insulin Release from
Incubated Rat Pancreatic Islets. The method used to measure
insulin release from incubated rat pancreatic islets was pre-
8) Babenko, A. P.; Gonzalez, G.; Bryan, J. Pharmacotopology of
Sulfonylurea Receptors. Separate Domains of the Regulatory
Subunits of KATP Channel Isoforms Are Required for Selective
a
values were calculated by the
þ
24
Interaction With K Channel Openers. J. Biol. Chem. 2000, 275,
7
17–720.
(
9) Moreau, C.; Prost, A. L.; D ꢀe rand, R.; Vivaudou, M. SUR, ABC
Proteins Targeted by KATP Channel Openers. J. Mol. Cell. Cardiol.
2
005, 38, 951–963.
1
3,20,25
viously described in detail.
(10) Garlid, K. D.; Paucek, P.; Yarov-Yarovoy, V.; Murray, H. N.;
Darbenzio, R. B.; D’Alonzo, A. J.; Lodge, N. J.; Smith, M. A.;
Grover, G. J. Cardioprotective Effect of Diazoxide and Its Inter-
Measurement of the Contractile Activity in Rat Aorta. The
methods used to measure the myorelaxant effect of the drugs
on 30 mM KCl-precontracted rat aorta rings was previously
described in detail.8
Measurement of Rb Outflow from Rat Pancreatic Islets. The
method used for measuring Rb ( K substitute) outflow from
prelabeled and perifused rat pancreatic islets was previously
described in detail.
Measurements of Ca Outflow and Insulin Release from
Perifused Islets. The method used for measuring Ca outflow
þ
action With Mitochondrial ATP-sensitive K Channels. Possible
Mechanism of Cardioprotection. Circ. Res. 1997, 1072–1082.
1
3,25
(
11) Pirotte, B.; de Tullio, P.; Lebrun, P.; Antoine, M. H.; Fontaine, J.;
Masereel, B.; Schynts, M.; Dupont, L.; Herchuelz, A.; Delarge, J.
3-(Alkylamino)-4H-pyrido[4,3-e]-1,2,4-thiadiazine 1,1-Dioxides
as Powerful Inhibitors of Insulin Release From Rat Pancreatic
B-cells: a New Class of Potassium Channel Openers? J. Med. Chem.
1993, 36, 3211–3213.
(12) de Tullio, P.; Becker, B.; Boverie, S.; Dabrowski, M.; Wahl, P.;
Antoine, M.-H.; Somers, F.; Sebille, S.; Ouedraogo, R.; Hansen,
J. B.; Lebrun, P.; Pirotte, B. Toward Tissue-selective Pancreatic
B-cells KATP Channel Openers Belonging to 3-Alkylamino-7-halo-
4H-1,2,4-benzothiadiazine 1,1-Dioxides. J. Med. Chem. 2003, 46,
6
8
6
42
20,26
4
5
4
5
from prelabeled and perifused rat pancreatic islets was pre-
viously described in detail.
2
0,26
Electrophysiology. HEK293 cells stably expressing human
SUR1 and human Kir6.2 channel were used in this study.
Whole-cell currents were recorded at 20-22 °C using an EPC9
patch-clamp amplifier and PulseþPulseFit v8.07 software. The
extracellular bath solution contained (in mM): 140 NaCl, 5 KCl,
3
342–3353.
(
13) de Tullio, P.; Boverie, S.; Becker, B.; Antoine, M.-H.; Nguyen,
Q. A.; Francotte, P.; Counerotte, S.; Sebille, S.; Pirotte, B.; Lebrun,
P. 3-Alkylamino-4H-1,2,4-benzothiadiazine 1,1-Dioxides as ATP-
sensitive Potassium Channel Openers: Effect of 6,7-Disubstitution
on Potency and Tissue Selectivity. J. Med. Chem. 2005, 48, 4990–
5000.
(14) Nielsen, F. E.; Bodvarsdottir, T. B.; Worsaae, A.; MacKay, P.;
Stidsen, C. E.; Boonen, H. C.; Pridal, L.; Arkhammar, P. O.; Wahl,
P.; Ynddal, L.; Junager, F.; Dragsted, N.; Tagmose, T. M.;
Mogensen, J. P.; Koch, A.; Treppendahl, S. P.; Hansen, J. B.
1
0 Hepes, 1.8 CaCl
Cells were dialyzed with intracellular solution containing (in
mM): 120 KCl, 1 MgCl , 5 EGTA, 2 CaCl , 20 Hepes (pH 7.3
with KOH), 3.0 K -ATP, and 0.3 K -ADP. Cells were clamped
2
, and 20 mannitol (pH 7.4 with NaOH).
2
2
2
2
at -80 mV and currents were evoked by repetitive 200 ms,
1
2
0 mV depolarizing voltage steps. Signals were sampled at
0 kHz and filtered at 5 kHz.
6-Chloro-3-alkylamino-4H-thieno[3,2-e]-1,2,4-thiadiazine 1,1-Di-
oxide Derivatives Potently and Selectively Activate ATP Sensitive
Potassium Channels of Pancreatic Beta-Cells. J. Med. Chem. 2002,
4
5, 4171–4187.
Acknowledgment. This study was supported by grants
(
15) Pirotte, B.; de Tullio, P.; Antoine, M.-H.; Sebille, S.; Florence, X.;
Lebrun, P. New insights into the development of ATP-sensitive
potassium channel openers. Expert Opin. Ther. Patents 2005, 15,
from Novo Nordisk and from the National Fund for Scien-
tific Research (FNRS, Belgium) from which P. de Tullio is a
Research Associate and P. Lebrun is a Research Director. The
authors gratefully acknowledge the technical assistance of
S. Counerotte, A.-M. Vanbellinghen, F. Leleux, and A. Van
Praet.
4
97–504.
(
16) Dabrowski, M.; Ashcroft, F. M.; Ashfield, R.; Lebrun, P.; Pirotte,
B.; Egebjerg, J.; Bondo Hansen, J.; Wahl, P. The Novel Diazoxide
Analog 3-Isopropylamino-7-methoxy-4H-1,2,4-benzothiadiazine
1
,1-Dioxide Is a Selective Kir6.2/SUR1 Channel Opener. Diabetes
2
002, 51, 1896–1906.
(
17) Dabrowski, M.; Larsen, T.; Ashcroft, F. M.; Bondo Hansen, J.;
Wahl, P. Potent and Selective Activation of the Pancreatic Beta-cell
Type KATP Channel by Two Novel Diazoxide Analogues. Diabe-
tologia 2003, 46, 1375–1382.
Supporting Information Available: Synthesis of compounds
1
and 25b, as well as elemental analyses of compounds 16, 17,
18a-d, 19a-e, 23, 24, and 25a-c. This material is available free
of charge via the Internet at http://pubs.acs.org.
2, 14, 16, 18a-d, 19a-d, 25a,c, all spectral data except for 19e
(
18) Raffa, L.; Di Bella, M.; Ferrari, P.; Rinaldi, M.; Ferrari, N. Azione
Cardiovasculare di Derivati dell’1,2,4-Benzothiadiazin-1,1-diossi-
do. Nota IV. Il Pharmaco Ed. Sci. 1974, 29, 411–423.
(
19) de Tullio, P.; Pirotte, B.; Lebrun, P.; Fontaine, J.; Dupont, L.;
Antoine, M. H.; Ouedraogo, R.; Khelili, S.; Maggetto, C.; Maser-
eel, B.; Diouf, O.; Podona, T.; Delarge, J. 3- and 4-Substituted 4H-
Pyrido[4,3-e]-1,2,4-thiadiazine 1,1-Dioxides as Potassium Channel
Openers: Synthesis, Pharmacological Evaluation, and Structure-
-Activity Relationships. J. Med. Chem. 1996, 39, 937–948.
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