Upload
others
View
6
Download
0
Embed Size (px)
Citation preview
Regioselective Palladium Catalyzed Cross-CouplingReactions of 2,4,7-Trichloroquinazoline and Progress
Towards Small-Molecule Inhibitors for Protein Kinase D
Kara GeorgeWipf Group Research Topic Seminar
5 June 2010
Kara George @ Wipf Group Page 1 of 78 12/5/2010
Regioselective Cross-Coupling Reactionsof 2,4,7-Trichloroquinazoline
N
N
Cl
Cl
Cl N
N
R3
R2
R1
Kara George @ Wipf Group Page 2 of 78 12/5/2010
Quinazoline Scaffold: ���Overview and Biological Significance
§ Quinazoline skeleton is present in a variety of biologically active compounds
- Among the most potent tyrosine kinase and cellular phosphorylation
inhibitors
- Exhibit remarkable activity as antitubercular, antiviral, antibacterial, and
anticancer agents
N
N
1
2
34
4a5
6
788a
Fry, D. W.; Kraker, A. J.; McMichael, A.; Ambroso, L. A.; Nelson, J. M.; Leopold, W. R.; Connors, R. W.; Bridges, A. J. Science. 1999, 265, 1093.
Portela-Cubillo, F.; Scott, J. S.; Walton, J. C. J. Org. Chem. 2009, 74, 4934.
NMeO
N
NH2
N
MeO
N
O
O
Prazosin
NO
N
HNOMeO
MeO
ErlotinibNMeO
N
HNO
Cl
F
NO
Iressa
Kara George @ Wipf Group Page 3 of 78 12/5/2010
Regioselective Pd-Catalyzed Cross-Couplingof 2,4,7-Trichloroquinazoline
Can we use consecutive Suzuki cross-coupling reactions to accesstricarbosubstituted quinazolines?
N
N
Cl
Cl
Cl N
N
R3
R2
R1
R1B(OH)2 R2B(OH)2 R3B(OH)2
Kara George @ Wipf Group Page 4 of 78 12/5/2010
Regioselective Pd-Catalyzed Cross-Couplingof 2,4,7-Trichloroquinazoline
N
N
Cl
Cl
Cl N
N
R3
R2
R1
R1B(OH)2 R2B(OH)2 R3B(OH)2
Can we use consecutive Suzuki cross-coupling reactions to accesstricarbosubstituted quinazolines?
Kara George @ Wipf Group Page 5 of 78 12/5/2010
Bond Dissociation Energy of Halo-Pyrimidines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position, then the 5-position
N
N
1
2
34
5
6N
N94.9
93.8Cl
Cl N
N96.5
99.6
Cl
Cl
N
N95.1Cl N
N93.7
Cl
N
N99.0Cl
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.
Kara George @ Wipf Group Page 6 of 78 12/5/2010
Bond Dissociation Energy of Halo-Pyrimidines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position, then the 5-position
Stille cross-coupling
N
N
1
2
34
5
6N
N94.9
93.8Cl
Cl N
N96.5
99.6
Cl
Cl
N
N95.1Cl N
N93.7
Cl
N
N99.0Cl
N
N
Cl
Cl
PdCl2(PPh3)2
DMF, 70 °C, 7 h
PhSnBu3
N
N
Cl
Ph
77%
PdCl2(PPh3)2
DMF, 130 °C, 15 h
PhSnBu3N
N
Ph
Ph
54%
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.Solberg, J.; Undheim, K. Acta Chem. Scand. 1989, 43, 62.
Kara George @ Wipf Group Page 7 of 78 12/5/2010
Bond Dissociation Energy of Halo-Pyrimidines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position, then the 5-position
Stille cross-coupling
N
N
1
2
34
5
6N
N94.9
93.8Cl
Cl N
N96.5
99.6
Cl
Cl
N
N95.1Cl N
N93.7
Cl
N
N99.0Cl
N
NCl
Cl
PdCl2(PPh3)2
DMF, 100 °C, 7 h N
NCl
Ph70%
PhSnBu3
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.Solberg, J.; Undheim, K. Acta Chem. Scand. 1989, 43, 62.
Kara George @ Wipf Group Page 8 of 78 12/5/2010
Bond Dissociation Energy of Halo-Pyrimidines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position, then the 5-position
Stille cross-coupling
N
N
1
2
34
5
6N
N94.9
93.8Cl
Cl N
N96.5
99.6
Cl
Cl
N
N95.1Cl N
N93.7
Cl
N
N99.0Cl
N
N
ClCl
ClN
NCl
Cl
Ph
71%
PdCl2(PPh3)2
DMF, 70 °C, 3 hPh
SnBu3
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.Solberg, J.; Undheim, K. Acta Chem. Scand. 1989, 43, 62.
Kara George @ Wipf Group Page 9 of 78 12/5/2010
Bond Dissociation Energy of Halo-Pyrimidines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position, then the 5-position
Suzuki cross-coupling
N
N
1
2
34
5
6N
N94.9
93.8Cl
Cl N
N96.5
99.6
Cl
Cl
N
N95.1Cl N
N93.7
Cl
N
N99.0Cl
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.Schomaker, J. M.; Delia, T. J. J. Org. Chem. 2001, 66, 7125.
N
N
Ph
ClCl N
N
Cl
ClCl N
N
Ph
ClPh
N
N
Ph
PhPh
5% Pd(OAc)2, 10% PPh3aq. Na2CO3, PhB(OH)2 (1 equiv)
DME, reflux, 18 h
88%
5% Pd(OAc)2, 10% PPh3aq. Na2CO3, PhB(OH)2 (2 equiv)
DME, 70 °C, 24 h
88%
93%
PhB(OH)2 (3.3 equiv)
Kara George @ Wipf Group Page 10 of 78 12/5/2010
Bond Dissociation Energy of Halo-Quinazolines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position All positions on the phenyl ring have similar C-Cl BDEs (96-97 kcal/mol) and 5-6
kcal/mol higher than the 2-position
N
N
1
2
345
6
78
N
N
94.6
91.996.397.3
96.996.9Cl
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.
Kara George @ Wipf Group Page 11 of 78 12/5/2010
Bond Dissociation Energy of Halo-Quinazolines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position All positions on the phenyl ring have similar C-Cl BDEs (96-97 kcal/mol) and 5-6
kcal/mol higher than the 2-position
Pd-catalyzed couplings of trialkylalanes
N
N
1
2
345
6
78
N
N
94.6
91.996.397.3
96.996.9Cl
N
N
Cl
Cl N
N
Me
Cl N
N
MePd(PPh3)4, AlMe3
THF, reflux
76% 80%
Pd(PPh3)4, Al(i-Bu)3
THF, reflux
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.Mangalagiu, I.; Benneche, T.; Undheim, K. Acta Chem. Scand. 1996, 50, 914.Mangalagiu, I.; Benneche, T.; Undheim, K. Tetrahedron Lett. 1996, 37, 1309.
Kara George @ Wipf Group Page 12 of 78 12/5/2010
Bond Dissociation Energy of Halo-Quinazolines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position All positions on the phenyl ring have similar C-Cl BDEs (96-97 kcal/mol) and 5-6
kcal/mol higher than the 2-position
Pd-catalyzed couplings of trialkylalanes
N
N
1
2
345
6
78
N
N
94.6
91.996.397.3
96.996.9Cl
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.Mangalagiu, I.; Benneche, T.; Undheim, K. Acta Chem. Scand. 1996, 50, 914.Mangalagiu, I.; Benneche, T.; Undheim, K. Tetrahedron Lett. 1996, 37, 1309.
N
N
Cl
Cl N
N
Me
Cl
Pd(PPh3)4, AlMe3
THF, reflux
47%
Br
N
N
Me
Cl16%
Br Me
Kara George @ Wipf Group Page 13 of 78 12/5/2010
Bond Dissociation Energy of Halo-Quinazolines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position All positions on the phenyl ring have similar C-Cl BDEs (96-97 kcal/mol) and 5-6
kcal/mol higher than the 2-position
Sonagashira cross-coupling
N
N
1
2
345
6
78
N
N
94.6
91.996.397.3
96.996.9Cl
N
N
Cl
Cl N
N
Cl
Bu
N
N
Bu
Bu
PdCl2(PPh3)2, CuI
NEt3, 20 °C, 20 h
H Bu 55%
65 °C
66%
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.Mangalagiu, I.; Benneche, T.; Undheim, K. Acta Chem. Scand. 1996, 50, 914.
Kara George @ Wipf Group Page 14 of 78 12/5/2010
Bond Dissociation Energy of Halo-Quinazolines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position All positions on the phenyl ring have similar C-Cl BDEs (96-97 kcal/mol) and 5-6
kcal/mol higher than the 2-position
Sonagashira cross-coupling
N
N
1
2
345
6
78
N
N
94.6
91.996.397.3
96.996.9Cl
N
N
Cl
Cl
Br
N
N
Cl
Bu
Br
N
N
Cl
Cl
BuPdCl2(PPh3)2, CuI
NEt3, 20 °C, 20 h
H Bu 67% 22%
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.Mangalagiu, I.; Benneche, T.; Undheim, K. Acta Chem. Scand. 1996, 50, 914.
Kara George @ Wipf Group Page 15 of 78 12/5/2010
Bond Dissociation Energy of Halo-Quinazolines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position All positions on the phenyl ring have similar C-Cl BDEs (96-97 kcal/mol) and 5-6
kcal/mol higher than the 2-position
Stille cross-coupling
N
N
1
2
345
6
78
N
N
94.6
91.996.397.3
96.996.9Cl
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.Charpiot, B.; Brun, J.; Donze, I.; Naef, R.; Stefani, M.; Mueller, T. Biorg. Med. Chem. Letters. 1998, 8, 2891.
N
N
Cl
Cl
O
O N
N
Cl
O
O
OBnBnO
BnO
BnO
SnBu3
PdCl2(PPh3)2, xylene110 °C, 3 h
Kara George @ Wipf Group Page 16 of 78 12/5/2010
Bond Dissociation Energy of Halo-Quinazolines:Relationship to Regioselectivity
Lowest C-Cl BDE at the 4-position, followed by the 2-position All positions on the phenyl ring have similar C-Cl BDEs (96-97 kcal/mol) and 5-6
kcal/mol higher than the 2-position
Consecutive Pd-catalyzed Suzuki cross-couplings
N
N
1
2
345
6
78
N
N
94.6
91.996.397.3
96.996.9Cl
Garcia, Y.; Schoenebeck, F.; Legault, C. Y.; Merlic, C. A.; Houk, K. N. J. Am. Chem. Soc. 2009, 131, 6632.
Cross-Coupling at C-2
Cross-Couplingat C-4
N
N
Cl
Cl
Cl N
N
Cl
R2
R1N
N
Cl
R2
Cl N
N
R3
R2
R1
Cross-coupling at C-7
R1B(OH)2R2B(OH)2 R3B(OH)2
Kara George @ Wipf Group Page 17 of 78 12/5/2010
Regioselective Suzuki Cross-Coupling at C-4
NH2Cl
OH
O
NCl
N
OH
OH NCl
N
Cl
Cl
H2O, 35% NaOH KOCN, 40 °C;
then AcOH, RT, 6 h
POCl3, N,N-dimethylaniline
reflux, 6 h
79% 56%
NCl
N
Cl
Cl
NCl
N
Cl
O5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, 80 °C, 16 h
86%
O
B(OH)21.1 equiv.
NCl
N
ClNCl
N
Cl NCl
N
ClNCl
N
Cl
S
71%73% 33%50%
Curd, F. H. S.; Landquist, J. K.; Rose, F. L. J. Chem. Soc. 1948, 1759.
Kara George @ Wipf Group Page 18 of 78 12/5/2010
Regioselective Suzuki Cross-Coupling at C-4
NH2Cl
OH
O
NCl
N
OH
OH NCl
N
Cl
Cl
H2O, 35% NaOH KOCN, 40 °C;
then AcOH, RT, 6 h
POCl3, N,N-dimethylaniline
reflux, 6 h
79% 56%
NCl
N
Cl
Cl
NCl
N
Cl
O5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, 80 °C, 16 h
86%
O
B(OH)21.1 equiv.
NCl
N
ClNCl
N
Cl NCl
N
ClNCl
N
Cl
S
71%73% 33%50%
Curd, F. H. S.; Landquist, J. K.; Rose, F. L. J. Chem. Soc. 1948, 1759.
Kara George @ Wipf Group Page 19 of 78 12/5/2010
Regioselective Suzuki Cross-Coupling at C-4
NH2Cl
OH
O
NCl
N
OH
OH NCl
N
Cl
Cl
H2O, 35% NaOH KOCN, 40 °C;
then AcOH, RT, 6 h
POCl3, N,N-dimethylaniline
reflux, 6 h
79% 56%
NCl
N
Cl
Cl
NCl
N
Cl
O5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, 80 °C, 16 h
86%
O
B(OH)21.1 equiv.
NCl
N
ClNCl
N
Cl NCl
N
ClNCl
N
Cl
S
71%73% 33%50%
Curd, F. H. S.; Landquist, J. K.; Rose, F. L. J. Chem. Soc. 1948, 1759.
Kara George @ Wipf Group Page 20 of 78 12/5/2010
Regioselective Suzuki Cross-Coupling at C-4
NH2Cl
OH
O
NCl
N
OH
OH NCl
N
Cl
Cl
H2O, 35% NaOH KOCN, 40 °C;
then AcOH, RT, 6 h
POCl3, N,N-dimethylaniline
reflux, 6 h
79% 56%
NCl
N
Cl
Cl
NCl
N
Cl
O5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, 80 °C, 16 h
86%
O
B(OH)21.1 equiv.
NCl
N
ClNCl
N
Cl NCl
N
ClNCl
N
Cl
S
71%73% 33%50%
Curd, F. H. S.; Landquist, J. K.; Rose, F. L. J. Chem. Soc. 1948, 1759.
Kara George @ Wipf Group Page 21 of 78 12/5/2010
Regioselective Suzuki Cross-Coupling at C-2
NCl
N
Cl
5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 22 h
O
NCl
N
O
CH3
B(OH)2
85%1.1 equiv.
NCl
N
O
S
77%
S B(OH)25% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 15 hNCl
N
Cl
O
1.1 equiv.
NCl
NCH3
S
45%
CH3
B(OH)2NCl
N
Cl
S5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 24 h
1.1 equiv.
Kara George @ Wipf Group Page 22 of 78 12/5/2010
Regioselective Suzuki Cross-Coupling at C-2
NCl
N
Cl
5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 22 h
O
NCl
N
O
CH3
B(OH)2
85%1.1 equiv.
NCl
N
O
S
77%
S B(OH)25% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 15 hNCl
N
Cl
O
1.1 equiv.
NCl
NCH3
S
45%
CH3
B(OH)2NCl
N
Cl
S5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 24 h
1.1 equiv.
Kara George @ Wipf Group Page 23 of 78 12/5/2010
Regioselective Suzuki Cross-Coupling at C-2
NCl
N
Cl
5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 22 h
O
NCl
N
O
CH3
B(OH)2
85%1.1 equiv.
NCl
N
O
S
77%
S B(OH)25% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 15 hNCl
N
Cl
O
1.1 equiv.
NCl
NCH3
S
45%
CH3
B(OH)2NCl
N
Cl
S5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 24 h
1.1 equiv.
Kara George @ Wipf Group Page 24 of 78 12/5/2010
Regioselective Suzuki Cross-Coupling at C-2
NCl
N
Cl
5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 22 h
O
NCl
N
O
CH3
B(OH)2
85%1.1 equiv.
NCl
N
O
S
77%
S B(OH)25% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 15 hNCl
N
Cl
O
1.1 equiv.
NCl
NCH3
S
45%
CH3
B(OH)2NCl
N
Cl
S5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 24 h
1.1 equiv.
Kara George @ Wipf Group Page 25 of 78 12/5/2010
Alternative Route to Sequential Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
Competitive hydrolysis at C-4
New Approach
NCl
N
S
Cl NCl
N
S
R1
OO
NCl
N
S
R1
[O]
Cross-Coupling at C-2
Cross-Couplingat C-4
Deactivation of C-4
N
N
Cl
Cl
Cl N
N
Cl
R2
R1N
N
Cl
R2
Cl N
N
R3
R2
R1
Cross-coupling at C-7
Cross-Coupling at C-2
Cross-Couplingat C-4
R1B(OH)2R2B(OH)2 R3B(OH)2
R1B(OH)2
N
N
Cl
OH
ClNCl
N
Cl
Cl
5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, 80 °C, 16 hB(OH)2
1.1 equiv.
NCl
N
Cl
33%
Kara George @ Wipf Group Page 26 of 78 12/5/2010
Alternative Route to Sequential Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
Competitive hydrolysis at C-4
New Approach
NCl
N
S
Cl NCl
N
S
R1
OO
NCl
N
S
R1
[O]
Cross-Coupling at C-2
Cross-Couplingat C-4
Deactivation of C-4
N
N
Cl
Cl
Cl N
N
Cl
R2
R1N
N
Cl
R2
Cl N
N
R3
R2
R1
Cross-coupling at C-7
Cross-Coupling at C-2
Cross-Couplingat C-4
R1B(OH)2R2B(OH)2 R3B(OH)2
R1B(OH)2
N
N
Cl
OH
ClNCl
N
Cl
Cl
5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, 80 °C, 16 hB(OH)2
1.1 equiv.
NCl
N
Cl
33%
Kara George @ Wipf Group Page 27 of 78 12/5/2010
Alternative Route to Sequential Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
Competitive hydrolysis at C-4
Initial Approach
N
N
Cl
OH
ClNCl
N
Cl
Cl
5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, 80 °C, 16 hB(OH)2
1.1 equiv.
NCl
N
Cl
33%
NCl
N
S
Cl NCl
N
S
R1
OO
NCl
N
S
R1
[O]
Cross-Coupling at C-2
Cross-Couplingat C-4
Deactivation of C-4
N
N
Cl
Cl
Cl N
N
Cl
R2
R1N
N
Cl
R2
Cl N
N
R3
R2
R1
Cross-coupling at C-7
Cross-Coupling at C-2
Cross-Couplingat C-4
R1B(OH)2R2B(OH)2 R3B(OH)2
R1B(OH)2
Kara George @ Wipf Group Page 28 of 78 12/5/2010
Alternative Route to Sequential Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
Competitive hydrolysis at C-4
New Approach
N
N
Cl
OH
ClNCl
N
Cl
Cl
5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, 80 °C, 16 hB(OH)2
1.1 equiv.
NCl
N
Cl
33%
NCl
N
S
Cl NCl
N
S
R1
OO
NCl
N
S
R1
[O]
Cross-Coupling at C-2
Cross-Couplingat C-4
Deactivation of C-4
N
N
Cl
Cl
Cl N
N
Cl
R2
R1N
N
Cl
R2
Cl N
N
R3
R2
R1
Cross-coupling at C-7
Cross-Coupling at C-2
Cross-Couplingat C-4
R1B(OH)2R2B(OH)2 R3B(OH)2
R1B(OH)2
Kara George @ Wipf Group Page 29 of 78 12/5/2010
Alternative Route to Sequential Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
NCl
N
S
NCl
N
Cl
Cl NCl
N
S
Cl
HSPh, NaH
THF, 0 °C, 18 h
75%
5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 23 h S
87%S(HO)2B
NCl
N
S
S mCPBA, CH2Cl20 °C - RT, 1.5 h
Na2WO4, H2O2MeOH, 50 °C, 24 h
Oxone, MeOH:H2O 0 °C - RT, 24 h
N
N
Cl
OH
NCl
N
S
S
OO
S
Kara George @ Wipf Group Page 30 of 78 12/5/2010
Alternative Route to Sequential Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
NCl
N
S
NCl
N
Cl
Cl NCl
N
S
Cl
HSPh, NaH
THF, 0 °C, 18 h
75%
5% Pd(OAc)2, 15% PPh3aq. Na2CO3
DME, reflux, 23 h S
87%S(HO)2B
NCl
N
S
S mCPBA, CH2Cl20 °C - RT, 1.5 h
Na2WO4, H2O2MeOH, 50 °C, 24 h
Oxone, MeOH:H2O 0 °C - RT, 24 h
N
N
Cl
OH
NCl
N
S
S
OO
S
Kara George @ Wipf Group Page 31 of 78 12/5/2010
Pd-Catalyzed, Cu-Mediated Cross-Coupling
ArB(OH)2cat. Pd, CuTC
THF, 50 °CHetAr SMe HetAr Ar
Pd (0)
PdL
L SR
Het
Pd SR Cu
O
OSB
O
R' OH
HHetL
L
PdL
L SR
Het
CuO
OS
PdL
L R'
Het
R'B(OH)2
CuTC
Het R' Het SR
Liebeskind, L. S.; Srogl, J. Org. Lett. 2002, 4, 979.
Kara George @ Wipf Group Page 32 of 78 12/5/2010
Catalytic Cycle:Pd-Catalyzed, Cu-Mediated Cross-Coupling
ArB(OH)2cat. Pd, CuTC
THF, 50 °CHetAr SMe HetAr Ar
Pd (0)
PdL
L SR
Het
Pd SR Cu
O
OSB
O
R' OH
HHetL
L
PdL
L SR
Het
CuO
OS
PdL
L R'
Het
R'B(OH)2
CuTC
Het R' Het SR
Liebeskind, L. S.; Srogl, J. Org. Lett. 2002, 4, 979.
Kara George @ Wipf Group Page 33 of 78 12/5/2010
Catalytic Cycle:Pd-Catalyzed, Cu-Mediated Cross-Coupling
ArB(OH)2cat. Pd, CuTC
THF, 50 °CHetAr SMe HetAr Ar
Pd (0)
PdL
L SR
Het
Pd SR Cu
O
OSB
O
R' OH
HHetL
L
PdL
L SR
Het
CuO
OS
PdL
L R'
Het
R'B(OH)2
CuTC
Het R' Het SR
Liebeskind, L. S.; Srogl, J. Org. Lett. 2002, 4, 979.
Kara George @ Wipf Group Page 34 of 78 12/5/2010
Catalytic Cycle:Pd-Catalyzed, Cu-Mediated Cross-Coupling
ArB(OH)2cat. Pd, CuTC
THF, 50 °CHetAr SMe HetAr Ar
Pd (0)
PdL
L SR
Het
Pd SR Cu
O
OSB
O
R' OH
HHetL
L
PdL
L SR
Het
CuO
OS
PdL
L R'
Het
R'B(OH)2
CuTC
Het R' Het SR
Liebeskind, L. S.; Srogl, J. Org. Lett. 2002, 4, 979.
Kara George @ Wipf Group Page 35 of 78 12/5/2010
Catalytic Cycle:Pd-Catalyzed, Cu-Mediated Cross-Coupling
ArB(OH)2cat. Pd, CuTC
THF, 50 °CHetAr SMe HetAr Ar
Pd (0)
PdL
L SR
Het
Pd SR Cu
O
OSB
O
R' OH
HHetL
L
PdL
L SR
Het
CuO
OS
PdL
L R'
Het
R'B(OH)2
CuTC
Het R' Het SR
Liebeskind, L. S.; Srogl, J. Org. Lett. 2002, 4, 979.
(OH)2BTCCuSR
Kara George @ Wipf Group Page 36 of 78 12/5/2010
Catalytic Cycle:Pd-Catalyzed, Cu-Mediated Cross-Coupling
ArB(OH)2cat. Pd, CuTC
THF, 50 °CHetAr SMe HetAr Ar
Pd (0)
PdL
L SR
Het
Pd SR Cu
O
OSB
O
R' OH
HHetL
L
PdL
L SR
Het
CuO
OS
PdL
L R'
Het
R'B(OH)2
CuTC
Het R' Het SR
Liebeskind, L. S.; Srogl, J. Org. Lett. 2002, 4, 979.
(OH)2BTCCuSR
Kara George @ Wipf Group Page 37 of 78 12/5/2010
Pd-Catalyzed, Cu-Mediated Cross-Coupling
ArB(OH)2cat. Pd, CuTC
THF, 50 °CHetAr SMe HetAr Ar
Efficient Pd-catalyzed, Cu-mediated coupling of heteroaromatic thioethers with boronicacids by Liebeskind and co-workers
Preparation of bis-fuctionalized pyridopyrimidine derivatives by Guillaumet and co-workers
N
N
Me
SMe
B(OH)2
NO2
4% Pd2(dba)3, 16% P(2-furyl)3CuTC (1.4 equiv), THF, 50 °C
N
N
Me
NO2
74%
N
N
N
Cl
Cl
N
N
N
S
Cl
N
N
N
R2
R1
N
N
N
S
R1
Deactivation of C-4
5% Pd(PPh3)4Na2CO3 (2 equiv)DME/H2O, 75 °C
R1B(OH)2
5% Pd(PPh3)4CuTC (2.2 eq)
THF, 50 °C
R2B(OH)2
Liebeskind, L. S.; Srogl, J. Org. Lett. 2002, 4, 979.Abdellatif, T.; Routier, S.; Akssira, M.; Leger, J.-M.; Jarry, C.; Guillaumet, G. Org. Lett. 2007, 9, 4673.
Kara George @ Wipf Group Page 38 of 78 12/5/2010
Pd-Catalyzed, Cu-Mediated Cross-Coupling
ArB(OH)2cat. Pd, CuTC
THF, 50 °CHetAr SMe HetAr Ar
Efficient Pd-catalyzed, Cu-mediated coupling of heteroaromatic thioethers with boronicacids by Liebeskind and co-workers
Preparation of bis-fuctionalized pyridopyrimidine derivatives by Guillaumet and co-workers
N
N
Me
SMe
B(OH)2
NO2
4% Pd2(dba)3, 16% P(2-furyl)3CuTC (1.4 equiv), THF, 50 °C
N
N
Me
NO2
74%
N
N
N
Cl
Cl
N
N
N
S
Cl
N
N
N
R2
R1
N
N
N
S
R1
Deactivation of C-4
5% Pd(PPh3)4Na2CO3 (2 equiv)DME/H2O, 75 °C
R1B(OH)2
5% Pd(PPh3)4CuTC (2.2 eq)
THF, 50 °C
R2B(OH)2
Liebeskind, L. S.; Srogl, J. Org. Lett. 2002, 4, 979.Abdellatif, T.; Routier, S.; Akssira, M.; Leger, J.-M.; Jarry, C.; Guillaumet, G. Org. Lett. 2007, 9, 4673.
Kara George @ Wipf Group Page 39 of 78 12/5/2010
A Third Approach:Pd-Catalyzed, Cu-Mediated Cross-Coupling
ArB(OH)2cat. Pd, CuTC
THF, 50 °CHetAr SMe HetAr Ar
New Approach:
Cross-Coupling at C-2
N
N
Cl
Cl
Cl
N
N
R3
R2
R1
Cross-coupling at C-7
Deactivation of C-4
N
N
S
ClCl N
N
S
R1Cl
N
N
R2
R1Cl
Pd-catalyzed, Cu-mediatedCross-Coupling at C-4
R1B(OH)2
R2B(OH)2 R3B(OH)2
Kara George @ Wipf Group Page 40 of 78 12/5/2010
Regioselective Pd-Catalyzed Cross-Coupling at C-2
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
CF3
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
S
NH
O NCl
N
S
O
NCl
N
S
N
99% 79%
66% 53%
92% 90% 95% 93%
89% 89%
NCl
N
Cl
Cl
i-PrSH, NaH
THF, 0 °C, 16 hNCl
N
S
Cl94%
5% Pd(OAc)2, 15% PPh3aq. Na2CO3, R1B(OH)2
DME, 75 °C NCl
N
S
R1
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 41 of 78 12/5/2010
Regioselective Pd-Catalyzed Cross-Coupling at C-2
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
CF3
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
S
NH
O NCl
N
S
O
NCl
N
S
N
99% 79%
66% 53%
92% 90% 95% 93%
89% 89%
NCl
N
Cl
Cl
i-PrSH, NaH
THF, 0 °C, 16 hNCl
N
S
Cl94%
5% Pd(OAc)2, 15% PPh3aq. Na2CO3, R1B(OH)2
DME, 75 °C NCl
N
S
R1
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 42 of 78 12/5/2010
Pd-Catalyzed, Cu-Mediated Cross-Coupling at C-4
NCl
N
NCl
N
NCl
N
NCl
NO NCl
NO
NCl
NS
CF3
NCl
N
O
89% 89% 86%
87% 97% 91%
83%
NCl
N
R1 NCl
N
R2
R1
S5% Pd(PPh3)4, CuTC (2.2 eq)
R2B(OH)2 (2.2 eq)
THF, 50 °C
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 43 of 78 12/5/2010
Pd-Catalyzed, Cu-Mediated Cross-Coupling at C-4
NCl
N
NCl
N
NCl
N
NCl
NO NCl
NO
NCl
NS
CF3
NCl
N
O
89% 89% 86%
87% 97% 91%
83%
NCl
N
R1 NCl
N
R2
R1
S5% Pd(PPh3)4, CuTC (2.2 eq)
R2B(OH)2 (2.2 eq)
THF, 50 °C
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 44 of 78 12/5/2010
Final Cross-Coupling at C-7
N
NNCl
N
B(OH)2
5% Pd(OAc)2, 15% PPh3aq. Na2CO3 (3.1 equiv)
DME, reflux, 24 h
50% conversion
O O
(2 equiv)
N
N
CF3
NCl
N
CF3
10% Pd(OAc)2, 30% PPh3aq. Na2CO3 (6.1 equiv)
DME, reflux, 34 h
81%
B(OH)2
(4 equiv)O O
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 45 of 78 12/5/2010
Final Cross-Coupling at C-7
N
NNCl
N
B(OH)2
5% Pd(OAc)2, 15% PPh3aq. Na2CO3 (3.1 equiv)
DME, reflux, 24 h
50% conversion
O O
(2 equiv)
N
N
CF3
NCl
N
CF3
10% Pd(OAc)2, 30% PPh3aq. Na2CO3 (6.1 equiv)
DME, reflux, 34 h
81%
B(OH)2
(4 equiv)O O
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 46 of 78 12/5/2010
Final Cross-Coupling at C-7
N
NNCl
N
B(OH)2
5% Pd(OAc)2, 15% PPh3aq. Na2CO3 (3.1 equiv)
DME, reflux, 24 h
50% conversion
O O
(2 equiv)
N
N
CF3
NCl
N
CF3
10% Pd(OAc)2, 30% PPh3aq. Na2CO3 (6.1 equiv)
DME, reflux, 34 h
81%
B(OH)2
(4 equiv)O O
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 47 of 78 12/5/2010
Final Cross-Coupling at C-7
N
NNCl
N
B(OH)2
5% Pd(OAc)2, 15% PPh3aq. Na2CO3 (3.1 equiv)
DME, reflux, 24 h
50% conversion
O O
(2 equiv)
N
N
CF3
NCl
N
CF3
10% Pd(OAc)2, 30% PPh3aq. Na2CO3 (6.1 equiv)
DME, reflux, 34 h
81%
B(OH)2
(4 equiv)O O
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 48 of 78 12/5/2010
Final Cross-Coupling at C-7
N
N
O
NCl
N10% Pd(OAc)2, 30% PPh3
aq. Na2CO3 (6.1 equiv)
DME, reflux, 22 h
O
B(OH)2
86%(4 equiv)
N
N
O
NCl
N
O
10% Pd(OAc)2, 30% PPh3aq. Na2CO3 (6.1 equiv)
DME, reflux, 34 h
66%(4 equiv)
NH
B(OH)2
NH
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 49 of 78 12/5/2010
Subsequent Regioselective Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
NCl
N
Cl
Cl
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 50 of 78 12/5/2010
Subsequent Regioselective Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
NCl
N
S
Cl
NCl
N
S
NCl
N
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NS
O
NH
CF3
O
NCl
N
NCl
N
NCl
NO
NCl
NO
NCl
NS
CF3
NCl
N
S
N
N
O
NCl
N
O
N
NO
CF3
SNCl
N
N
N
O
NH
NCl
N
O
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 51 of 78 12/5/2010
Subsequent Regioselective Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
NCl
N
S
Cl
NCl
N
S
NCl
N
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NS
O
NH
CF3
O
NCl
N
NCl
N
NCl
NO
NCl
NO
NCl
NS
CF3
NCl
N
S
N
N
O
NCl
N
O
N
NO
CF3
SNCl
N
N
N
O
NH
NCl
N
O
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 52 of 78 12/5/2010
Subsequent Regioselective Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
NCl
N
S
Cl
NCl
N
S
NCl
N
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NS
O
NH
CF3
O
NCl
N
NCl
N
NCl
NO
NCl
NO
NCl
NS
CF3
NCl
N
S
N
N
O
NCl
N
O
N
NO
CF3
SNCl
N
N
N
O
NH
NCl
N
O
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 53 of 78 12/5/2010
Subsequent Regioselective Cross-CouplingReactions of 2,4,7-Trichloroquinazoline
NCl
N
S
Cl
NCl
N
S
NCl
N
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NCl
N
S
NS
O
NH
CF3
O
NCl
N
NCl
N
NCl
NO
NCl
NO
NCl
NS
CF3
NCl
N
S
N
N
O
NCl
N
O
N
NO
CF3
SNCl
N
N
N
O
NH
NCl
N
O
Wipf, P.; George, K. M. Synlett. 2010, 644.
Kara George @ Wipf Group Page 54 of 78 12/5/2010
Introduction: Protein Kinase D
PKD1: PKD/PKCµ
PKD2
PKD3: PKDυ (PKCυ)
PKD
Constitutes a novel family of serine/threonine kinases and diacylglycerol(DAG) receptors that belong to the Ca2+/calmodulin-dependent kinase(CaMK) superfamily
Three PKD isoforms: PKD1 (PKCµ), PKD2, PKD3 (PKCυ) PKD has been linked to a diverse set of signal transduction pathways
involved in tumor development and cancer progression
AP
P
C1 AC PH KinaseC1a C1b
S
C1a C1b
C1a C1b
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.For reviews see: Rozengurt, E.; Rey, O.; Waldron, R. T. J. Bio. Chem 2005, 280, 13205. LaValle, C. R.; George, K. M.; Sharlow, E. R.; Lazo, J. S.; Wipf, P.; Wang, Q. J. BBA Rev. Cancer. 2010, in press
Kara George @ Wipf Group Page 55 of 78 12/5/2010
Introduction: Protein Kinase D
PKD1: PKD/PKCµ
PKD2
PKD3: PKDυ (PKCυ)
PKD
AP
P
C1 AC PH KinaseC1a C1b
S
C1a C1b
C1a C1b
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
O
O
H3C(H2C)12
CH3
CH3O
O
CH3
H
H
OH
HH3C
H3C
O HO
OH
phorbol ester (PMA)
OHO
O R1
OO
R2
Diacylglycerol (DAG)
Autoinhibition
Protein-ProteinInteractions
PKCs
P P
Kara George @ Wipf Group Page 56 of 78 12/5/2010
Introduction: Protein Kinase D
PKD1: PKD/PKCµ
PKD2
PKD3: PKDυ (PKCυ)
PKD
AP
P
C1 AC PH KinaseC1a C1b
S
C1a C1b
C1a C1b
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
O
O
H3C(H2C)12
CH3
CH3O
O
CH3
H
H
OH
HH3C
H3C
O HO
OH
phorbol ester (PMA)
OHO
O R1
OO
R2
Diacylglycerol (DAG)
Autoinhibition
Protein-ProteinInteractions
PKCs
P P
Kara George @ Wipf Group Page 57 of 78 12/5/2010
Introduction: Protein Kinase D
PKD1: PKD/PKCµ
PKD2
PKD3: PKDυ (PKCυ)
PKD
AP
P
C1 AC PH KinaseC1a C1b
S
C1a C1b
C1a C1b
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
O
O
H3C(H2C)12
CH3
CH3O
O
CH3
H
H
OH
HH3C
H3C
O HO
OH
phorbol ester (PMA)
OHO
O R1
OO
R2
Diacylglycerol (DAG)
Autoinhibition
Protein-ProteinInteractions
c/nPKCs
P P
Kara George @ Wipf Group Page 58 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 59 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLC
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 60 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLCDAG
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 61 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLCnPKC
DAG
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 62 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLCnPKCPKD
DAG
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 63 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLCnPKCPKD
p p DAG
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 64 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLCnPKCPKD
p p DAG
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 65 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLCnPKCPKD
p p DAG
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 66 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLCnPKCPKD
p p DAG
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 67 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLCnPKCPKD
p p DAG
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 68 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLCnPKCPKD
p p DAG
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
ROS
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
Kara George @ Wipf Group Page 69 of 78 12/5/2010
Protein Kinase D Signaling Pathway
Wang, Q. J. Trends in Pharmacol. Sci. 2006, 27, 317.
PLCnPKCPKD
p p DAG
PKD
p p
Mitochondria Nucleus
Stimuli
Receptors
II. Regulation of class IIa HDAC
IV. Oxidative stress response
ERK1/2
I. Modulation of signaling pathways
RIN1
p14-3-3
Ras
Raf
MEK
MEKK
MKK
JNK
c-Junp
III. Membrane orprotein trafficking
TGN
PKD
p p
PKCηGbγ
PI4K IIIβp
PKD
p pp
PKCδ
Src
AbI
PKD
p p
Oxidative Stress
IKKcomplex
MnSODNF-κB
HDAC IIa
p14-3-3
HDAC IIaHDAC IIa
pPKD
p p
MEF2
14-3-3
X
ROS
Kara George @ Wipf Group Page 70 of 78 12/5/2010
Chemical Inhibitors of PKD
ATP competitive Staurosporine and staurosporine-derived compounds
Gschwendt, M.; Dieterich, S.; Rennecke, J.; Kittstein, W.; Mueller, H.-J.; Johannes, F.-J. FEBS Letters 1996, 392, 77.Martiny-Baron, G.; Kazanietz, M. G.; Mischak, H.; Blumberg, P. M.; Kochs, G.; Hug, H.; Marme, D.; Schachtele, C. J. Biol. Chem. 1993, 268, 9194.Zang, R.; Mueller, H.-J.; Kielbassa, K.; Marks, F. Biochem. J. 1994, 304, 641.
Abbreviation: n.d., not determined
HNO
N NOCH3OCH3
HN CH3
Staurosporine
HN OO
N NH
O
NGö 6983
HNO
N NCH3 CNGö6976
HNO
N NOCH3
CO2CH3OH
K252a
PKC!
330 nM7 nM
2.3 nM 17 nM
Inhibitor
StaurosporineGö 6983Gö 6976K252a
PKC"
430 nM6 nM n.d.
18 nM
PKC#
>10,000 nM10 nM
>10,000 nM 3000 nM
PKC$
330 nM7 nM
6.2 nM 15 nM
PKC%
>10,000 nM 60 nM
>10,000 nM 10 nM
PKCµ/PKD
40 nM20,000 nM
20 nM7 nM
Kara George @ Wipf Group Page 71 of 78 12/5/2010
Chemical Inhibitors of PKD
ATP competitive Resveratrol
In vitro IC50 (PKD) = 52 - 200 μM In vivo IC50 (PKD) = 760 μM
Stewart, J. R.; Christman, K. L.; O’Brian, C. A. Biochem. Pharmacol. 2000, 60, 1355.Haworth, R. S.; Avkiran, M. Biochem. Pharmacol. 2001, 62, 1647.
HO
OH
OH
Resveratrol
Kara George @ Wipf Group Page 72 of 78 12/5/2010
First Potent and Selective Inhibitor of PKD: CID755673
Non-competitive with respect to ATP and substrate Effective at blocking PKD-mediated cell functions Reduced cell migration, invasion, and proliferation in prostate cancer cells
CID755673
O
NHHO
O
Kinase
PKD1PKD2PKD3PLK1CAKAKT
PKC!PKC"
CAMPK!
IC50 (µM)
0.5 ± 0.030.3 ± 0.0020.2 ± 0.0220.3 ± 10.915.3 ± 1.8 18.6 ± 2.0
not significantno activity40.5 ± 4.6
Sharlow, E. R.; Giridhar, K. V.; LaValle, C. R.; Chem, J.; Leimgruber, S.; Barrett, R.; Bravo-Altamirano, K.; Wipf, P.; Lazo, J.; Wang, Q. J. J. Biol.Chem. 2008, 283, 33516.
Kara George @ Wipf Group Page 73 of 78 12/5/2010
First Potent and Selective Inhibitor of PKD: CID755673
Cellular activity relatively weak: IC50 = 10 - 30 µM
May possess unintended PKD-independent effects
CID755673
O
NHHO
O
Torres-Marquez, E.; Sinnett-Smith, J.; Guha, S.; Kui, R.; Waldron, R. T.; Rey, O.; Rosengurt, E. Biochem. Biophys. Res. Comm. 2010, 391, 63.
Kara George @ Wipf Group Page 74 of 78 12/5/2010
Chemical Inhibitors of PKD
Monovich, L.; Vega, R. B.; Meredith, E.; Miranda, K.; Rao, C.; Capparelli, M.; Lemon, D. D.; Phan, D.; Koch, K. A.; Chapo, J. A.; Hood, D. B.; McKinsey, T.A. FEBS Letters 2010, 584, 631.
PKD1PKD2PKD3CaMKI!CaMKII"CaMKII#CaMKII!CaMKIVMARK1MARK2SIK1GRK5PKC!PKC$
IC50 (µM)
10210210960326124277222-3
Kinase % Inhibition at1µM BPKDi
0.0010.0090.001n.d.n.d.n.d.n.d.n.d.n.d.n.d.n.d.n.d.n.d.n.d.
N
N
H2N
O
NH
N
NH
BPKDi
Blocks signal-dependent phosphorylation and nuclear export of class IIaHDACs in cardiomyocytes, thereby suppressing hypertrophy of these cells
ATP competitive
Kara George @ Wipf Group Page 75 of 78 12/5/2010
Chemical Inhibitors of PKD
Harikumar, K. B.; Kunnumakkara, A. B.; Ochi, N.; Tong, Z.; Deorukhkar, A.; Sung, B.; Kelland, L.; Jamieson, S.; Sutherland, R.; Raynham, T.; Carles, M.;Bagherazadeh, A.; Foxton, C.; Boakes, A.; Farooq, M.; Maru, D.; Diagaradjane, P.; Matsuo, Y.; Sinnett-Smith, J.; Gelovani, J.; Krishnan, S.; Aggarwal, B. B.;Rozengurt, E.; Ireson, C. R.; Guha, S. Mol. Cancer Ther., 2010, 9, 1136.
CRT0066101
N
N
NR2
R1 NH2
HOR3
First orally bioavailable PKD inhibitor Blocks pancreatic cancer growth both in vitro and in vivo Specificity confirmed in an in vitro assay comprisising a panel of >90 protein
kinases ATP competitive
Kinase
PKD1PKD2PKD3
IC50 (µM)
0.0010.0030.002
Kara George @ Wipf Group Page 76 of 78 12/5/2010
Conclusion and Future Work
Methoxy derivative successfully synthesized and biological testing is inprogress
Optimize deprotection and submit for biological testing
Photoaffinity labeled derivative to gain insight into binding site
Kara George @ Wipf Group Page 77 of 78 12/5/2010
Acknowledgements
Dr. Wipf Dr. Wang, Courtney LaValle, Dr. Karla Bravo-Altamirano Wipf group members past & present NMR & MS facilities NIH, University of Pittsburgh Arts & Science Fellowship
Kara George @ Wipf Group Page 78 of 78 12/5/2010