SAGE-217: A 2nd Generation Neuroactive Steroid Albert J. Robichaud, James J. Doherty, Francesco G. Salituro, Gabriel Martinez-Botella, Ethan Hoffmann,

Rebecca S. Hammond, Gabriel M. Belfort, Mike A. Ackley SAGE Therapeutics, Cambridge, MA

• Seizure disorders that remain resistant to current therapies represent a significant unmet medical need

• γ-Containing receptors are present predominantly at synaptic sites, whereas δ-containing receptors populate extra-synaptic sites

• Benzodiazepines bind to the interface of the α and γ subunits and so do not modulate δ-containing receptors. In contrast neuroactive steroids (NASs) bind to the α subunit and so can potentiate a broader range of GABAA receptors

• First generation NASs, e.g. ganaxolone and allopregnanolone, are limited with regard to the route of administration due to their DMPK and physical properties.

• Second generation NASs, like SAGE-217, are designed to modulate GABAA receptors while achieving the right balance of physical properties and DMPK suitable for oral and i.v. dosing.

• Here we describe the properties of our 2nd generation NAS, SAGE-217 in comparison to 1st generation NASs allopregnanolone and ganaxolone

Overview

• We have developed a second generation series of novel neuroactive steroid compounds with a range of pharmacologies and pharmacokinetic properties suitable for oral administration.

• SAGE-217 is a potent and highly selective GABAA receptor PAM. It has a formulation and pharmacokinetic profile differentiated from 1st Gen NAS; the compound is predicted to be suitable for either oral or parenteral dosing

• SAGE-217 produces 1) dose-dependent anticonvulsant activity and 2) arrest of pharmaco-resistant SE in preclinical models

• SAGE-217 is an NCE with patent applications filed for composition of matter, formulation and utilities claims

Conclusions

SAGE-217 is potent and efficacious at both synaptic and extrasynaptiuc GABAA receptors

Superior selectivity vs 1st generation NASs

Key properties Comparison

Acknowledgments: The authors would like to extend their thanks to Carla Maciag, Scott J. Grossman and Boyd L. Harrison, Shanghai Chempartners, and Wuxi

Synaptic Extrasynaptic

5

-5

0 EE

G (m

V)

SAGE-217 (5 mpk iv.)

SAGE-217 Aborts SE (60 min post-onset) Diazepam Fails to Abort SE (60 min post-onset)

Piloca

rpine

SE Onse

t 1 2 3 4 5

-1

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Rela

tive

EEG

Pow

er (3

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Vehicle 1 mg/kg 3 mg/kg 5 mg/kg

SAGE-217

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*

DZ (10 mpk iv.)

5

-5

0

Time (h)

0 1 2 3 4 5

Time (h)

0 1 2 3 4 5

DZ

Piloca

rpine

SE Onse

t 1 2 3 4 5

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tive

EEG

Pow

er (3

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Vehicle 10 mg/kg

EEG

(mV)

Vehicle SAGE-547 SAGE-217 Ganaxolone

SE onset

Dose SE onset Dose SE onset Dose

SE onset Dose

SE onset Dose

SE onset Dose

5854±448 ng/ml 5657±2482 ng/ml

SE onset

Dose SE onset Dose

2088±132 ng/ml

1.0

10.0

100.0

1000.0

10000.0

0 2 4 6 8 10 12 14

[Con

c], n

g/m

L

Time (hr)

Ganaxolone, 20 mg/kg PO

SAGE-547, 20 mg/kg PO

SAGE-217, 10 mg/kg PO

PK parameters Unit Ganaxolone SAGE-547 SAGE-217

Tmax hr 1.00 0.500 0.500

Cmax ng/mL 60.1 13.0 1335

Terminal t1/2 hr 3.65 NA 1.98

AUClast-plasma hr*ng/mL 111 9.16 4055

AUCINF-plasma hr*ng/mL 127 NA 4096

F % 4.31 0.563 62.4

SAGE-217 SAGE-547 Ganaxolone

a1 EC50 (nM)/Emax 652/641 149/343 440/397

a4 EC50 (nM)/Emax 317/531 80/418 100/219

LogD/Aq Sol. (µM) 4.6/2 4.9/3.3 5.3/0.7

Microsomes M/R/D/H Clint (µl/min/mg) 77/44/9/6 415/992/38/16 251/85/31/12

B/P (mouse, rat) 1.5, 3.2 1.9, 1.8 2.5, 4.1

Mouse PO F% / t1/2 (h) 62/2.0 0.5/ND 4/3.6

Rat PO F% / t1/2 (h) 53/2.0 2.3/2.5 10/2.1

Dog PO F% / t1/2 (h) 68/12.9 ND ND

PTZ at MED Tonic+%Death (mpk/nM) (dose/ plasma/brain) 0.3 (IP)/186/204 10 (IP)/453/ND 10 (IP)/1327/ND

Li-Pilo SE model (50% seizure termination 60 mins post onset)

mpk/nM 3-4 mpk iv 10 mpk iv No effect (>12 mpk iv)

Nuclear Hormone Receptor Affinity All >10 UM AR, PPAR, PROG, ERb FXR, AR, RORg, PROG(ant)

A h rA RC A R 3

E R aE R ß

F X R G RL X R a

L X R ß

P P A R a

P P A R d

P P A R g P RP X R

R A R aR A R b

R A R gR X R a

T R aT R ß

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fN H R a g o n is tS A G E -2 1 7

Fold

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ivatio

n

4 0 0 nM

2uM

1 0uM

A h rA RC A R 3

E R aE R ß

F X R G RL X R a

L X R ß

P P A R a

P P A R d

P P A R g P RP X R

R A R aR A R b

R A R gR X R a

T R aT R ß

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fN H R a n ta g o n is tS A G E -2 1 7

%in

hibi

tion

4 0 0 nM

2uM

1 0uM

A h rA RC A R 3

E R aE R ß

F X RG RL X R a

L X R ßP P A R a

P P A R d

P P A R g P RP X R

R A R aR A R b

R A R gR X R a

T R aT R ß

0

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6 0

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fN H R a g o n is tG a n a x o lo n e

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ation

4 0 0 nM

2uM

1 0uM

A h r A RC A R 3

E R aE R ß

F X R G RL X R a

L X R ßP P A R a

P P A R d

P P A R g P RP X R

R A R aR A R b

R A R gR X R a

T R aT R ß

0

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8 0

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fN H R a n ta g o n is tG a n a x o lo n e

%inh

ibitio

n

4 0 0 nM

2uM

1 0uM

A h rA RC A R 3

E R aE R ß

F X R G RL X R a

L X R ß

P P A R a

P P A R d

P P A R g P RP X R

R A R aR A R b

R A R gR X R a

T R aT R ß

0

2 0

4 0

6 0

8 0

1 0 0

fN H R a g o n is tS A G E -5 4 7

Fold

Activ

ation

4 0 0 nM

2uM

1 0uM

A h rA RC A R 3

E R aE R ß

F X R G RL X R a

L X R ß

P P A R a

P P A R d

P P A R g P RP X R

R A R aR A R b

R A R gR X R a

T R aT R ß

0

2 0

4 0

6 0

8 0

1 0 0

fN H R a n ta g o n is tS A G E -5 4 7

%inh

ibitio

n

4 0 0 nM

2uM

1 0uM

fNHR agonist SAGE-217

fNHR antagonist SAGE-217

fNHR agonist SAGE-547

fNHR agonist GANAXOLONE

fNHR antagonist SAGE-547

fNHR antagonist GANAXOLONE

SAGE-217: Dose dependent termination of pharmacoresistant SE

SAGE-217 SAGE-547 Ganaxolone

EC50 (nM)

SAGE-217 188

SAGE-547 452

Ganaxolone 1270

Active in PTZ model with optimal sedation profile

http://www.sagerx.com/posters/poster5.pdf

α1 Synaptic

α4 Extrasynaptic

EC50 (nM) Emax (%)

EC50 (nM)

Emax (%)

SAGE-217 652 641 317 531

SAGE-547 149 342 80 418

Ganaxolone 440 397 100 219