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0.001 0.01 0.1 1 100
10000
20000
30000
40000 ND-630 CP-640186 (3μM)
cpm
ND-630 (μM)
0.001
0.01
0.1
1
10
Plasma Liver Muscle
Cm
ax (μ
M)
0
10
20
30
40
50
60
Vehicle 0.03 0.1 0.3 1 3
0
1
2
3
Vehicle 0.3 3 300
1
2
3
4
5
6
Vehicle 0.3 3 30 Mal
onyl
-CoA
(nm
ol/g
Tis
sue)
mg/kg
mg/kg mg/kg
Mal
onyl
-CoA
(nm
ol/g
Tis
sue)
DPM
s/m
g Ti
ssue
0.7
0.8
0.9
1
1.1
1.2
-60 0 60 120 180 240
RQ
Time (min)
Vehicle 3 mg/kg 10 mg/kg 30 mg/kg
0.01
0.1
1
10
100
Plasma Liver Muscle
Tiss
ue
Con
cent
ratio
n (μ
M)
0
23
6
PBS 0.3 3 30mg/kg
1
54
Mal
onyl
-CoA
(n
mol
/g ti
ssue
)
0
4
5
Vehicle 0.3 3 30mg/kg
1
2
3
Mal
onyl
-CoA
(n
mol
/g ti
ssue
)
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
ICAM
-1/C
D54
CD26/D
pp4
IL-1B
IL-11
RANTES
IL-6
IL-10
MCP-
1/CCL2
Pref-
1/DLK
-1
RAGE
TNF-α LIF
0
1
2
3
4
5
6
TIMP-
1
IGFB
P1
IGFB
P2
IGFB
P3
IGFB
P5
IGFB
P6
FGF-
21
0
2
4
6
8
10
12
14
16
Serp
in 1
Endoca
n
Leptin
Resist
in
Fold
Exp
ress
ion
as C
ompa
red
to P
BS
Con
trol
PBS
DEN
DEN+10mpk ND-654
DEN+30mpk ND-654
KKeeyy
0.00015
0
0.0002
0.0003
0.00025
0.0001
0.0005
10
ND-654 (mg/kg)
30PB
S
met D
EN 10
ND-654 (mg/kg)
30PB
S
met D
EN
0.00015
0
0.0002
0.0003
0.00025
0.0001
0.0005
6
0
8
12
10
4
2
Are
a A
ffect
ed (%
)
10
ND-654 (mg/kg)
30PB
S
met D
EN
50mg/kg DEN IP, once a week
13 wk 18 wk sac
ND-654, PO, QD
8-weeks 12-weeks 18-weeks
Inflammation Fibrosis HCC
0
20
40
60
80
100
120
140
160
PBS DEN
Trig
lyce
rides
(mg/
dL)
DEN
P
Ser222
Arg277
Unstructured region of A-domain tail
0
100
200
300
400
500
600
700
800
Plas
ma-
Free
Fat
ty A
cids
Lean DIO 3 10 30
ND-630 (mg/kg) Vehicle
020406080
100120140160180
Lean DIO 3 10 30
ND-630 (mg/kg) Vehicle
0
20
40
60
80
100
120
Plas
ma
Cho
lest
erol
(mg/
dL)
Lean DIO 3 10 30
ND-630 (mg/kg) Vehicle
Plas
ma
Trig
lyce
rides
(mg/
dL)
0
ND-630 (mg/kg)
0.5 1.5 5
Live
r Fre
e Fa
tty
Aci
ds (n
mol
/mg)
0
20
15
10
5
*
Live
r Cho
lest
erol
(μg/
mg)
4
0
5
9876
123
*
0
ND-630 (mg/kg)
0.5 1.5 5Live
r Trig
lyce
rides
(μg/
mg)
0
40
30
20
10
*
0
ND-630 (mg/kg)
0.5 1.5 5
****** ***
***
*** ***
OTHERS
COO-
Acetyl CoA Malonyl CoA
BC CT
CPT1
Fatty Acids
ACC2
Mitochondrion
ACC1
Inhibitor
Acetyl-CoA
+ HCO3-
Malonyl-CoA
-CoA -CoA
Fatty acid synthesis
Tissue triglycerides
Body fat and weight
Fatty acid oxidation
Insulin sensitivity
ACC Inhibition Leads to:
Lipotoxic Metabolites
(ceramides, DAGs, lysophosphatidyl
choline, etc.)
Peroxisomal -oxidation
Lipid droplets
(steatosis)
Mitochondrial -oxidation
Triglyceride
VLDL (secreted)
• ER stress• Inflammation• Apoptosis• Necrosis
Hepatic Free Fatty Acids
Increased circulating fatty acids
Insulin resistance
Diet / uncontrolled diabetes
Lipotoxicity Hypothesis Excess peripheral
lipolysis
NASH
SER (P450) -oxidation
De novo lipogenesis
Excess carbohydrates
ACC Inhibitor
Increases -oxidation
ACC inhibitor reduces hepatic FFA’s and thereby lipotoxic metabolites both
by a) inhibiting de novo lipogenesis and b)
increasing -oxidation
de novo Lipogenesis
Steatosis Inflammation/Apoptosis Fibrosis/Cirrhosis
ND-630ND-654
Liver: Muscle 100:1
ND-654Liver : Muscle
2700:1
12-27%2
Simple Steatosis
NASH
Normal Liver
Cirrhosis HepatocellularCarcinoma
ND-630
OVERVIEW 1. Lipotoxicity as a Molecular Cause of NASH: ACC Inhibition Reduces FFA’s & Resulting Lipotoxicity1 3. Nimbus Solves ACC Druggability Challenge by Targeting Allosteric Sitein Biotin Carboxylase Domain
A B
C D
5. ND-630: Hepatotropic ACC Inhibitor 6. ND-630 Modulates Key Metabolic Parameters In Liver and Muscle
8. ND-654: Hepatoselective ACC Inhibitor
7. ND-630 Favorably Modulates Key Plasma and Liver Lipids in-vivo
9. DEN-Induced Model of Cirrhosis in Rat 10. ND-654 Shows Improvements in Fibrosis, Stellate Cell Activation and InflammationMarkers in the Rat DEN Model
11. Quantitative Liver Adipokine Array From ND-654 Treated DEN Rats DemonstratesACC Inhibition Modulates Fibrosis and Inflammation
2. Acetyl CoA Carboxylase (ACC) Regulates Fatty Acid Synthesis & Oxidation
4. Liver Health: Unmet Need in Nonalcoholic Steatohepatitis
Liver-Directed Allosteric Inhibitors of Acetyl-CoA Carboxylase Reduce Hepatic Steatosis and Improve Dyslipidemia in Diet-Induced Obese Rat Models and Reduce Inflammation and Fibrosis in a Cirrhotic Rat Model
Geraldine Harriman1, Danielle K. DePeralta2, Omeed Moaven2,Lan Wei2, Jeremy R. Greenwood3, Sathesh P. Bhat3, Kenneth K. Tanabe2, Bryan C. Fuchs2, William Westlin1, H. James Harwood1, Rosana Kapeller1
1Nimbus Therapeutics, Cambridge, MA, USA; 2Division of Surgical Oncology, Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, MA, USA;3Schrodinger, New York, NY
25 First St #404, Cambridge, MA 02141, USA • www.nimbustx.com • Tel: 857.999.2009 • [email protected]:
• Liver disease progression from Nonalcoholic Fatty Liver Disease (NAFLD) to Nonal-coholic Steatohepatitis (NASH) and Hepatocellular Carcinoma (HCC) has been welldocumented
• Hepatoselective (ND-630) and Hepatospecific (ND-654) inhibitors of Acetyl-CoA Car-boxylase (ACC) have been developed to address the continuum of these liver dis-eases
• These potent and selective allosteric inhibitors demonstrate excellent PK-PD rela-tionships in target tissues (exposure vs target engagement and efficacy) and are ef-fective at modulating NASH-relevant endpoints across several in vivo models
• ND-630 and ND-654 demonstrate the ability for tissue targeted ACC inhibition toimprove metabolic syndrome endpoints, decrease liver steatosis, decrease expres-sion of inflammatory markers and improve fibrosis
ND-630 was evaluated in three rat models of target engagement. (A,B) ND-630 shows a dose-dependent reduction in the formation of the enzymatic product of acetyl coAcarboxylase, malonyl coA. This reduction occurs in both the liver and muscle tissues. In alignment with the hepatoselective nature of the bio-distribution of ND-630, the ED50 in muscle proved to be lower. (C,D) ND-630 demonstrates its effectiveness at inhibiting the production of fatty acids in the liver (C, ED50 = 0.14 mg/kg) and modulating respiratoryquotient (D, MED = 3 mg/kg).
• Only potent BC domain inhibitor:Soraphen A
• Industry efforts on analogingsoraphen proved unsuccessful
• Allosteric binding site implicatedin dimerization of ACC
1. Novel class of ACC inhibitors
2. Prototype for modulating protein-proteininteractions
Nimbus inhibitors prevent ACC dimerization and activation
ND-630 binds to the same site as Soraphen A
Nimbus BreakthroughChallenges in Drugging ACC
Target potency• ACC1 IC50 = 2 nM• ACC2 IC50 = 7 nM• HepG2 EC50 = 9 nM (serum free)
= 66 nM (10% serum)• C2C12 FAOxn stim. = 2x @ 200 nM
Pharmacology• Rat FASyn ED50 = 0.14 mpk PO• Rat RQ MED = 3 mpk PO• Rat Malonyl-CoA ED50 = 0.8 mg/kg (liver)• Rat Malonyl-CoA ED50 = 3 mg/kg (muscle)
ADME• Low multispecies intrinsic clearance (human,
mouse, rat, dog, monkey)• Eliminated predominantly as parent• P450 inhib >50 μM• Protein binding (98%)
ND-630
Hep-G2 Cell FA Synthesis [14C]acetate incorporation into FA
Exposure at 3 mg/kg
Liver Muscle
Decreases Fatty Acid Synthesis Increases Fatty Acid Oxidation
Decrease Liver Free Fatty Acids Decrease Plasma Triglycerides Decrease Plasma Cholesterol
Data are mean values for n=8 animals DIO, n=10 animals ZDF ± SEM
Target potency• ACC1 IC50 = 3 nM• ACC2 IC50 = 8 nM• HepG2 EC50 = 4 nM (serum free)
= 14 nM (10% serum)
Pharmacology• Rat FASyn ED50 = 0.3 mg/kg PO• Rat Malonyl-CoA ED50 = 0.3 mg/kg (liver)• Rat DEN HCC MED = 10 mg/kg
ADME• Moderate multispecies
intrinsic clearance(human, mouse, rat, dog, monkey)
• P450 inhib >50 μM• Highly selective across broad panel (>1000x)
ND-654Exposure at 10 mg/kg
1. Graphic adapted from Neuschwander-Tetri et al. Hepatology, 2010; 52:774-788.2. Arthur J. McCullough, Cleveland Clinic at the AASLD - FDA Workshop on NASH, Sep 5, 20132. AASLD Practice Guidelines for NAFLD, 2012
3. Schuppan et al., Liver International, 20104 US Census Bureau, 2013 estimate: 317 million 5. Venook et al. The Oncologist, 2010;15(suppl 4):5–13
DEN DEN +ND-654 10PBS
Collagen 1A1 marker (COL1A1)
(Fibrosis)
Sirius Red StainingAlpha-Smooth Muscle Actin (α-SMA)
(Stellate Cell Activation)
DEN +ND-654 30
α-S
MA
Inflammation Fibrosis Others
Liver
Muscle(Gastrocnemius)
p < 0.01
n.s.
Poster #957
6. Will develop NASH-related HCC over time7. Graphic adapted from Cohen et al. Science, 2011;332(6037): 1519–1523
• Diethyl-nitrosamine (DEN): powerful inducer of injury-repair process, inflammation, fibrosis, andoncogenesis in rodents
Recapitulates the histological changes of human NASH
• Weekly administration of DEN in rats causes progressive inflammation and fibrosis followed bydevelopment of cirrhosis by 18 weeks
• Highly lipophilic binding site• Inhibitors bind to active site• Past programs exhibited poor
drug-like properties
BC Domain CT Domain
• Beneficial effects on lipids, blood glucose, weight neutral• Nimbus: first allosteric inhibitor successfully targeting BC domain
• ACC inhibition directly addressescausative and perpetuatingbiological steps in NASH
Decrease Liver Free Fatty Acids Decrease Liver Triglycerides Decrease Liver Cholesterol
28d HS DIO Rat Endpoints
37d ZDF Rat Endpoints