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 • info@nimbustx.comNotes:

• 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