ROS as targets for therapeutic intervention of diabetic ... · 36 th Autumn Congress of Korean...
Transcript of ROS as targets for therapeutic intervention of diabetic ... · 36 th Autumn Congress of Korean...
36th Autumn Congress of Korean Diabetes AssociationSeptember 16-17 2010 BEXCO Busan Korea
ROS as targets for therapeutic intervention of diabetic nephropathy
Hunjoo Ha
Department of Bioinspired ScienceDivision of Life amp Pharmaceutical Sciences
College of Pharmacy Ewha Womans University Seoul Korea
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Contents
renal injury under hyperglycemiaRole of antioxidant defense mechanism
under hyperglycemia catalase Nrf2 Recent therapeutic advances supporting
the role of ROS in diabetic renal injury
O2 O2- H2O2 OH + OH- 2H2O
e e e e
4e
Sequential reduction of O2
O2-
Weak oxidant acting on Fe-S Fe-Zn centersH2O2 Weak oxidant oxidizing sulfur atoms of Cys and Met
OH Strong oxidant indiscriminately damaging DNA Proteins Lipids
O2 2H2O4e
+ 4H+
HO O
H
Finkel T et al Nature 408239 2000
O2-
H2O2
OH
Oxidative Stress
SODGPxcatalaseAntioxidantDefense
MechanismsROS
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Increased ROS under hyperglycemia
Koya D et al JASN 14S250 2003Ha H et al JASN 3894 2002 Unpublished data
Markers of OS in human DN
GFAT
protein
PKC
ROS-regulated signaling in renal cells under hyperglycemia
Lee HB et al JASN 14S241-5 2003
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Contents
renal injury under hyperglycemiaRole of antioxidant defense mechanism
under hyperglycemia catalase Nrf2 Recent therapeutic advances supporting
the role of ROS in diabetic renal injury
O2 O2- H2O2 OH + OH- 2H2O
e e e e
4e
Sequential reduction of O2
O2-
Weak oxidant acting on Fe-S Fe-Zn centersH2O2 Weak oxidant oxidizing sulfur atoms of Cys and Met
OH Strong oxidant indiscriminately damaging DNA Proteins Lipids
O2 2H2O4e
+ 4H+
HO O
H
Finkel T et al Nature 408239 2000
O2-
H2O2
OH
Oxidative Stress
SODGPxcatalaseAntioxidantDefense
MechanismsROS
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Increased ROS under hyperglycemia
Koya D et al JASN 14S250 2003Ha H et al JASN 3894 2002 Unpublished data
Markers of OS in human DN
GFAT
protein
PKC
ROS-regulated signaling in renal cells under hyperglycemia
Lee HB et al JASN 14S241-5 2003
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
O2 O2- H2O2 OH + OH- 2H2O
e e e e
4e
Sequential reduction of O2
O2-
Weak oxidant acting on Fe-S Fe-Zn centersH2O2 Weak oxidant oxidizing sulfur atoms of Cys and Met
OH Strong oxidant indiscriminately damaging DNA Proteins Lipids
O2 2H2O4e
+ 4H+
HO O
H
Finkel T et al Nature 408239 2000
O2-
H2O2
OH
Oxidative Stress
SODGPxcatalaseAntioxidantDefense
MechanismsROS
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Increased ROS under hyperglycemia
Koya D et al JASN 14S250 2003Ha H et al JASN 3894 2002 Unpublished data
Markers of OS in human DN
GFAT
protein
PKC
ROS-regulated signaling in renal cells under hyperglycemia
Lee HB et al JASN 14S241-5 2003
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Finkel T et al Nature 408239 2000
O2-
H2O2
OH
Oxidative Stress
SODGPxcatalaseAntioxidantDefense
MechanismsROS
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Increased ROS under hyperglycemia
Koya D et al JASN 14S250 2003Ha H et al JASN 3894 2002 Unpublished data
Markers of OS in human DN
GFAT
protein
PKC
ROS-regulated signaling in renal cells under hyperglycemia
Lee HB et al JASN 14S241-5 2003
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
O2-
H2O2
OH
Oxidative Stress
SODGPxcatalaseAntioxidantDefense
MechanismsROS
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Increased ROS under hyperglycemia
Koya D et al JASN 14S250 2003Ha H et al JASN 3894 2002 Unpublished data
Markers of OS in human DN
GFAT
protein
PKC
ROS-regulated signaling in renal cells under hyperglycemia
Lee HB et al JASN 14S241-5 2003
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Increased ROS under hyperglycemia
Koya D et al JASN 14S250 2003Ha H et al JASN 3894 2002 Unpublished data
Markers of OS in human DN
GFAT
protein
PKC
ROS-regulated signaling in renal cells under hyperglycemia
Lee HB et al JASN 14S241-5 2003
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Increased ROS under hyperglycemia
Koya D et al JASN 14S250 2003Ha H et al JASN 3894 2002 Unpublished data
Markers of OS in human DN
GFAT
protein
PKC
ROS-regulated signaling in renal cells under hyperglycemia
Lee HB et al JASN 14S241-5 2003
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Markers of OS in human DN
GFAT
protein
PKC
ROS-regulated signaling in renal cells under hyperglycemia
Lee HB et al JASN 14S241-5 2003
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
GFAT
protein
PKC
ROS-regulated signaling in renal cells under hyperglycemia
Lee HB et al JASN 14S241-5 2003
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
A heme-containing homotetrameric protein Mainly located in the peroxisomes
H2O2 O2 + 2H2O
OH- + OH
Catalase
Fe (Ⅱ)Fenton reaction
Role of catalase in diabetic nephropathy
Fenton reaction
Ho et al JBC 27932804 2004
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Characteristics of diabetic cat null mice
04050607
(mgd
L)
C
10
15
20
25
D
Pla
sm
a F
FA (m
M) dagger
B
Rela
tive
incre
ase
3456
daggersect
A
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Unpublished data
00102030405
PC
r (m
gd
L)
00
05
10
Pla
sm
a F
FA (m
M)
Rela
tive
incre
ase
0123
WT
WT+
DM
Cat
--
Cat
--+
DM
Cat --
WT Cat --+DMWT+DM
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
0
05
1
15
WT WT+DM Cat-- Cat--+DM
Relative increase
Glomerular volume
Fractional mesangial area
daggersect
Glomerular volume amp FMA in diabetic cat null mice
WT
Unpublished data
05
07
09
WT WT+DM Cat-- Cat--+DM
FMA
Fractional mesangial area (FMA)
Cat--
con DM
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Renal injury in diabetic cat null mice
00 05 10 15 20 25 30 35
TGF-
β1
18S
daggersect
A
00 05 10 15 20 25 30 35 40 45
CTG
F18
S
daggersect
B
00 05 10 15 20 25 30 35 40 45 50
BM
P7
18S
dagger
C
0
50
100
150
200
250
300
daggersect
BM
P7
(pgm
g p
rote
in)D
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat-- sect plt005 vs WT+DM
WT Cat-- Cat--+DM
D
BM
P7
(pgm
g p
rote
in)
WT+DM
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Oxidative stress in diabetic cat null mice
00
05
10
15
20
25
Nox4
18S
+
08
10
12
18S
WT
WT+
DM
plt005 vs WT dagger plt005 vs Cat--
00
03
05
08
10
13
15
SOD
218
S
00
02
04
06
08
Cat
alas
e18
S
WT+
DM
Cat
--
Cat
--D
M
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Nrf2-Keap1-ARE
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Diabetic renal injury in Nrf2 null mice
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Role of ROS in HG-induced Nrf2 activation
NAC
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Definition of the term ROS (reactive oxygen species) and OS (oxidative stress)
Current understanding of ROS-induced renal injury under hyperglycemia
Role of antioxidant defense mechanism Role of antioxidant defense mechanism under hyperglycemia catalase Nrf2
Recent therapeutic advances supporting the role of ROS in diabetic renal injury
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Dose of antioxidants Concentration of antioxidants at the site of ROS produced (eg mitochondria) Plasma half-life of antioxidants
Successful antioxidant therapy will depend onhellip
Plasma half-life of antioxidants Safety and tolerability of antioxidants Type of antioxidants antioxidants may act as oxidants Oxidative state of patients
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Bardoxolone methyl an inducer of Nrf2
Phase IIa trialRenoprotective effect of bardoxolone methyl on diabetes
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Selective mitochondrial antioxidants MitoQ
Permeable into mitochondria 100~1000 fold accumulation
Adsorbed to the matrix surface of inner membrane Accumulated in the heart liver skeletal muscle kidney and brain
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Oral bioavailability 10 Major metabolites in the urine glucuronide amp sulfate conjugation
PROTECT study Parkinsonrsquos diseasebull Double-blind studybull 40 80 mgkg for 12 months
Human studies with MitoQ
bull 40 80 mgkg for 12 months CLEAR trial chronic HCV unresponsive to conventional txbull Double-blind randomized studybull 0 40 80 mgkg for 28 days
Common adverse effect dose-dependent nausea
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Haptoglobin (Hp) genotype amp DN
a hemoglobin binding protein protects against heme-induced ROS 3 genotypes Hp 1-1 Hp 2-2 and Hp 2-1 Diabetes with Hp 2-2 have 2~5 fold increasedrisk of CVD
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Increased generation of ROS in diabetic kidney is convincing Endogenous antioxidant defense mechanism plays a protective role in DN
Antioxidants acting directly at the mitochondrial site may
Conclusion
Antioxidants acting directly at the mitochondrial site may promise better efficiency than non-specific antioxidants
Pharmacogenomic application may provide a rational for prospective trials to investigate the potential of vigorous antioxidant therapy to ameliorate DN
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Body wt (g)
Kidney wtbody wt (ratio)
Blood glucose (mgdL)
Plasma creatinine (mgdL)
UAE(mg24 hrs)
WT 228 plusmn 07 743 plusmn 008 193 plusmn 15 138 plusmn 027 060 plusmn 028
WT+DM 190 09 756 019 362 50 119 026 150 046
Characteristics of diabetic cat null mice
WT+DM 190 plusmn 09 756 plusmn 019 362 plusmn 50 119 plusmn 026 150 plusmn 046
Cat-- 275 plusmn 05 722 plusmn 006 175 plusmn 17 144 plusmn 025 048 plusmn 017
Cat--+DM 231 plusmn 09daggersect 784 plusmn 010dagger 397 plusmn 74dagger 114 plusmn 021 724 plusmn 265daggersect
n=7~8 Mean plusmn SE plt005 vs WTdagger plt005 vs Cat-- sect plt005 vs WT+DMUAE urinary albumin excretion
Unpublished data
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Renal injury in diabetic cat null mice
00 05 10 15 20 25
αSM
A1
8Sdaggersect
E
4
5
6
18S
G
E-cadherin
β-actin
α-SMA
Contr
ol
DM
Contr
ol
DM
WT Cat --
H
00
05
10
15
20
25
αSM
Aβ
-act
inI
20
25
cadher
in
actin
J
Unpublished datan=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
0
1
2
3
4
E-ca
dher
in1
8S
dagger
WT Cat
00
05
10
15
20
dagger
E-ca
dher
inβ-a
ctin
WT Cat-- Cat--+DM
D
WT+DM
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Oxidative stress in diabetic cat null mice
10
15
20
25
Nox4
18S
+
05
08
10
13
15
SOD
218
S
04
06
08
10
12
Cat
alas
e18
S
Unpublished data
n=7~8 Mean plusmn SE plt005 vs WT dagger plt005 vs Cat--
00
05 Nox4
18S
00
03 SOD
218
S
00
02
04
Cat
alas
e
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
Selective mitochondrial antioxidants MitoQ
httpwwwantipodeanpharmacom
httpwwwantipodeanpharmacom