Praliciguat, a clinical-stage sGC stimulator, improved ...

Praliciguat, a clinical-stage sGCstimulator, improved insulin sensitivity, lipid tolerance and energy utilization in a diet-induced obesity mouse model

Chad D. Schwartzkopf, John R. Hadcock, Julien Roux, Guang Liu, Courtney Shea, Mark G. Currie, G. Todd Milne, Jaime L.

Masferrer and Juli E. Jones

The current prevalence of obesity is alarming

2

WHOBody Mass Index (BMI) = kg/m2

• In 2014, an estimated 900 million worldwide were obese and 1.5 billion were overweight

• Obesity is associated with numerous comorbid conditions such as diabetes, cardiovascular disease and cancer

Clinical experience with the sGC stimulator praliciguat suggests potential metabolic benefit

3

• Praliciguat (IW-1973) is currently in Phase 2 for diabetic nephropathy and HFpEF

• In an exploratory Phase 2 study in 26 patients with type 2 diabetes and hypertension on standard of care therapy, 14 days of praliciguat treatment showed positive trends in reducing

• Fasting plasma glucose

• HOMA-IR

• LDL cholesterol

• Triglycerides

Ø The purpose of the current studies was to explore the mechanism of action behind the potential metabolic benefits of praliciguat utilizing an obese mouse model

Change in HOMA-IR (%)patients not on concomitant insulin therapy

-6 0

-4 0

-2 0

0

2 0

LS

Me

an

(9

5%

CI)

P la c e b o (n = 4 )

P ra lic ig u a t 4 0 m g (n = 1 2 )

-1 3 ( -4 0 , 1 5 ) -3 6 ( -5 3 , -1 9 )

L S M e a n D iffe re n c e (9 5 % C I)-2 3 ( -5 6 , 9 )

NCT03091920

Praliciguat shows extensive distribution to key target tissues

4

Heart Liver Kidney LungSkeletal Muscle

Adipose

Tissue/plasma Cmax ratio 5.0 53.0 9.6 4.1 3.1 12.5

Enhanced NO-sGC-cGMP signaling by praliciguat demonstrates benefits in preclinical models

5Adapted from Tobin, Zimmer et al. J Pharmacol Exp Ther 2018;365:664-675. Copyright © 2018 The Author(s).

• In preclinical models, compared to disease controls, animals treated with praliciguat have:

• Preserved cardiac function• Less cardiac hypertrophy• Lowered biomarkers of

inflammation and fibrosis• Less renal damage

Diet-induced obesity (DIO) mouse model is a common preclinical model of insulin resistance

• Simple model: C57Bl/6 mice are switched to 60% high fat diet (HFD) at 6 weeks of age

• Within 3 weeks mice develop an obese phenotype• Increased adiposity

• Insulin resistance

• Leptin resistance

• HFD is continued throughout study and animals will continue to gain weight

• Widely used in anti-obesity and metabolic syndrome researcho Animals will lose weight on anorectic drugs (GLP-1, phentermine)

o Animals will increase insulin sensitivity/secretion when on anti-diabetic drugs (GLP-1, metformin, rosiglitazone)

Study Design

7

• Study was performed at thermoneutrality (30° C)

• Housing animals below their thermoneutral zone alters energy expenditure and substrate utilization

• Cold stress undermines mouse modeling (Karp, J. Exp. Med. Vol. 209 No. 6 1069-1074 (2012))

Age = 6wk 12wk 14wk 18wk Collect blood, tissues after

overnight fast and 2H post PO

dose

Acclimation, T=30°C

60% HFD Pretreatment (8 weeks)

Maintained on LFD (8 weeks)

Obese Controls

PRL (6 mg/kg) in HFD

Study Phase

Praliciguat did not alter body weight, food intake or body composition

8

7 14 21 28 35

20

25

30

35

40

45

50

Study Day

Bod

y W

eigh

t, g

7 14 21 28 35

0

10

20

30

40

50

Study Day

Fat M

ass,

% o

f tot

al

7 14 21 28 35

40

50

60

70

80

Study Day

Lean

Mas

s, %

of t

otal

7 14 21 28 35

0

100

200

300

400

500

Study Day

Cum

ulat

ive

Calo

ricIn

take

, kca

l

LeanObesePRL

Body Weight Fat Mass Lean Mass Food Intake

Praliciguat-treated mice had greater insulin sensitivity than control DIO mice

• Compared to obese mice, four weeks of praliciguat resulted in:o Lower fasting insulino Lower C-peptide o Lower HOMA-IR

• In this model, fasting glucose was not altered in obese or PRL-treated mice

9

Insulin Glucose

C-Peptide HOMA-IR

Lean Obese PRL 0

1

2

3

4

Insu

lin, n

g/m

l

****

*

Lean Obese PRL 100

200

300

400

Glu

cose

, mg/

dl

Lean Obese PRL 0

1

2

3

4

C-P

eptid

e, n

g/m

l

****

*

Lean Obese PRL 0

10

20

30

40

50

****

HO

MA-

IR

*

DIO mice treated with praliciguat had lower expression of genes involved in inflammation

10

Liver: Tnf

Lean Obese PRL 0.000

0.005

0.010

0.015

0.020

norm

aliz

ed g

ene

expr

essi

on

****

**

Liver: TNFα SKM: Ccl2

Lean Obese PRL 0.00

0.02

0.04

0.06

0.08

norm

aliz

ed g

ene

expr

essi

on

**

*

Muscle: Ccl2

WAT: Akt1

Lean Obese PRL 0.0

0.5

1.0

1.5

2.0

2.5

norm

aliz

ed g

ene

expr

essi

on

**** *

WAT: Icam1

Lean Obese PRL 0.0

0.1

0.2

0.3

0.4

0.5

norm

aliz

ed g

ene

expr

essi

on

****

Adipose: Akt1 Adipose: Icam1

DIO mice treated with praliciguat had normalized expression of genes involved in lipid handling

11

Liver: Pnpla3

Lean Obese PRL 0.000

0.001

0.002

0.003

norm

aliz

ed g

ene

expr

essi

on

** *

Liver: Lpl

Lean Obese PRL 0.0

0.1

0.2

0.3

norm

aliz

ed g

ene

expr

essi

on

***

Liver: Pnpla3 Liver: Lipoprotein lipaseLiver: Slc27a1

Lean Obese PRL 0.0

0.2

0.4

0.6

0.8

norm

aliz

ed g

ene

expr

essi

on

*****

Liver: Long-chain fatty acid transport protein 1

SKM: Lipe

Lean Obese PRL 0.0

0.5

1.0

1.5

2.0

2.5

norm

aliz

ed g

ene

expr

essi

on

**

Muscle: Hormone-sensitive lipase WAT: Fdft1

Lean Obese PRL 0

1

2

3

4

norm

aliz

ed g

ene

expr

essi

on******

WAT: Ppara

Lean Obese PRL 0.0

0.1

0.2

0.3

norm

aliz

ed g

ene

expr

essi

on ****

**

Adipose: Farnesyl-diphosphate farnesyltransferase 1

Adipose: Pparα

DIO mice treated with praliciguat had lower expression of leptin in liver and muscle suggesting repartitioning of adipose tissue

12

Liver: Lep

Lean Obese PRL 0.000

0.005

0.010

0.015

norm

aliz

ed g

ene

expr

essi

on

***

**

Liver Leptin SKM: Lep

Lean Obese PRL 0

1

2

3

4

norm

aliz

ed g

ene

expr

essi

on

* *

Muscle Leptin

Nor

mal

ized

Gen

e Ex

pres

sion

The assessment of praliciguat on energy expenditure and lipid handling

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• Energy expenditure (EE) was measured on day 8, 9, 20, 21, 32 and 33

• A lipid tolerance test (LTT) was performed on day 38

Age = 6wk 12wk 14wk 18wk

Acclimation, T=30°C

60% HFD Pretreatment (8 weeks)

Maintained on LFD (8 weeks)

Obese Controls

PRL (6 mg/kg) in HFD

Study Phase

19wk

EE EE EE LTT

Praliciguat-treated mice had a mild increase in energy expenditure compared to obese controls

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• Increase in energy expenditure was accompanied with an increase in fat oxidation through day 21.

1:00

2:00

3:00

4:00

5:00

6:00

7:00

8:00

9:00

10:0

011

:00

12:0

013

:00

14:0

015

:00

16:0

017

:00

18:0

019

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20:0

021

:00

22:0

023

:00

200

250

300

350

400

Day 8

Hour

Ener

gy e

xpen

ditu

re(K

cal/d

ay/k

g le

an)

LeanObesePRL

Obese vs. Lean nsObese vs. PRL ****

1:00

2:00

3:00

4:00

5:00

6:00

7:00

8:00

9:00

10:0

011

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12:0

013

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015

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017

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019

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021

:00

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023

:00

200

250

300

350

400

Day 20

HourEn

ergy

exp

endi

ture

(Kca

l/day

/kg

lean

)

LeanObesePRL

Obese vs. Lean nsObese vs. PRL ****

1:00

2:00

3:00

4:00

5:00

6:00

7:00

8:00

9:00

10:0

011

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250

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Day 32

Hour

Ener

gy e

xpen

ditu

re(K

cal/d

ay/k

g le

an)

LeanObesePRL

Obese vs. Lean ***Obese vs. PRL ****

DIO mice treated with praliciguat had lower plasma triglycerides content suggesting improved lipid handling

15

Lean Obese PRL0

50

100

150

Trig

lyce

rides

, mg/

dl *¯13%**¯19%

0

100

200

300

400

ObesePRL

Time, hours

Trig

lyce

rides

, mg/

dl

Lean****

**

**

-0.5 51.5 3

Olive Oil10 µl/g

Oral Lipid Tolerance TestFasting Plasma Triglycerides

PKG VASP NF-kB

Inflammation

Insulin resistance

Ectopic fat accumulationMetabolic syndrome

Type 2 diabetesCardiovascular disease

High Fat Diet

Insulin Receptor

Inflammation and NO insufficiency impair insulin signaling

16

1. The consumption of high-fat diet can lead to obesity resulting in reduced NO signaling

2. Reductions in VASP and high fat diet both increase NF-kB, which increases inflammation and inhibits insulin signaling leading to insulin resistance

3. All of these factors lead to comorbid metabolic conditions

1

2

3

PKG VASP NF-kB

Inflammation

Insulin resistance

Ectopic fat accumulationMetabolic syndrome

Type 2 diabetesCardiovascular disease

High Fat Diet

Insulin Receptor

Praliciguat’s effect on inflammation and insulin sensitivity in DIO mice may be due to inhibition of NF-kB activation

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1. NO-sGC-cGMP signaling stimulates VASP phosphorylation which inhibits NF-kB

2. Praliciguat-treated mice had lower levels of inflammatory gene expression across target tissues

3. The increased insulin sensitivity in praliciguat-treated mice may be due to its effects on the NF-kB signaling pathway

1

32

Improved lipid handling in praliciguat-treated DIO mice may be in part due to the increase in adipose Pparα

18

1

2

Pparα

High Fat Diet

↓Triglycerides↑FFA Oxidation

1. High fat diet inhibits Pparα; yet praliciguat-treated mice had a restoration in adipose tissue pparα gene expression

2. Pparα decreases triglycerides and increases FFA oxidation

3. Dietary nitrate enhance Pparα and Pparβ/δ activity

4. Pparα agonists such as fenofibrate have been demonstrated to increase NO via eNOS

4

3

Summary

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§ The positive trends in several metabolic parameters observed in humans following praliciguat treatment is recapitulated in DIO mice including reduced fasting insulin, HOMA-IR, and fasting triglycerides

§ Praliciguat did not affect plasma glucose, body weight, food intake or body composition

§ Praliciguat treated-mice had positive changes in the expression of genes associated with inflammation and lipid metabolism in key metabolic tissues

§ The effects of praliciguat on inflammation and insulin sensitivity may be explained by inhibition of NF-kB signaling

§ The effects of praliciguat on lipid handling may be mediated by activation of Pparα

§ These data demonstrate broad metabolic effects such as improved insulin sensitivity, lipid handling and increased energy utilization in obese mice housed at thermoneutrality

Questions?

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