RO7020531, a novel prodrug of a toll-like receptor 7 agonist,

demonstrated desirable pharmacodynamics responses in both

AAV-HBV mice and healthy volunteers

Lu Gao1, Lue Dai1, Youjun Yu1, Xue Zhou1, Lili Gu1*, Yuyan Jin1, Yonghong Zhu1, Joseph F. Grippo2, Miriam Triyatni3, Ilia

Folitar3*, Ruchi Upmanyu4, Katerina Glavini3, Eoin Coakley2*, Tomas Racek3, Edward J. Gane5

1. Roche Innovation Centre Shanghai, China, 2. Roche Innovation Center New York, USA; 3. Roche Innovation Center Basel, Switzerland; 4. Roche Innovation Center Welwyn, UK; 5.

Auckland Clinical Studies, New Zealand; *Former Roche employee

Cure

Virus Targeted Approach

CpAM Class I RO7049389

VIRAL CYCLE

Immunoenhancers

TLR7 Agonist RO7020531

ActivationViral-mediated

suppression

Peg-IFN

Existing SoC

Virus Targeted asset

Immunoenhancer asset

Functional impairment of anti-HBV immune responses is a key feature of chronic HBV infection.

Development of a finite HBV cure will likely require combinations of novel compounds which inhibit HBV replication,

reduce antigen production and enhance HBV-specific immune responses2

HBV LNA

Nucleos(t)ide

Roche Approach Toward Combination Therapy for HBV Cure

Novel molecule

Novel molecule

CpAM: core protein allosteric modulator; LNA: locked nucleic acid; SoC: standard of care; IFN: interferon; TLR: toll-like receptor

Roche TLR7 agonist is positioned to be differentiated as an oral

double prodrug selectively converted in the liver

• TLR7 agonist induces broad immuno-modulatory effects including:

– Activation of IRF7, NFκB, and AP-1 transcription factors

– Differentiation of plasmacytoid dendritic cells (pDCs), and upregulation of co-stimulatory molecules and secretion of type 1-IFN and other

cytokines/chemokines leading to activation of T-cells and NK cells

– Differentiation of B-cells to antibody-producing plasma cells

• RO7020531 is an orally available double pro-drug of a TLR7 agonist which also activates TLR8 with lower potency

• It was safe and well tolerated in healthy volunteers with a favorable PK profile in single and multiple QOD doses up to 170 mg

Hydrolysis

Esterase

Oxidation

Aldehyde Oxidase

hTLR7 reporter EC50: 55.2 ±22.5 M

hTLR8 reporter EC50: 296 ±45 M

PBMC MEC IFNα induction: 3-6 M

CC50: > 1000 M

RO7020531

Double pro-drugSingle pro-drug Active form

Dai, L. et al. [poster]. EASL 2018; SAT-345; Gane, E. et al. [poster]. EASL 2018; FRI-3373

In the AAV-HBV mouse model, oral combination treatment with TLR7

agonist RO7020531 and CpAM RO7049389 leads to sustained viral

load suppression and HBsAg loss

4

Vehicle

TLR7 agonist 100

mg/kg QOD

CpAM

20 mg/kg QD

Combo

(n=7)

200

150

100

50

14 21 28 35 42 49 56 63 70 77 84Days

mIU

/ml

Treatment end

0

6

5

4

3

2

0 7 14 21 28 35 42 49 56 63 70 77 84

Days

Lo

g10

IU/m

l se

rum

Treatment end

LLOQ

*

Dai et al EASL 2018; SHC 2018, GHS 2018

• The combination of RO7049389 and RO7020531 reduced HBsAg level to below LLOQ at the end of treatment in 5 of 7 animals, which sustained

during 6-week off-treatment follow-up.

Results are presented as meanSEM (n = 7). LLOQ = lower limit of quantification; QD = once a day; QOD = every other day

Log 1

0co

pie

s/m

l ser

um

10

9

8

7

6

5

0 7 14 21 28 35 42 49 56 63 70 77 84Days

Treatment end

LLOQ

HBV DNA HBsAg Anti-HBs antibody

RO7020531 increases the number of germinal center B cells and

HBsAg-specific B and T cells in the spleen of AAV-HBV infected mice

• HBsAg-specific B cells were captured and measured by ELISPOT with HBsAg-coated plate.

• HBsAg-specific T cells were measured by IFN- γ ELISPOT in the presence of HBsAg peptides.

Weeks

-4 4-3 -2 0-1 1 2 3

AAV-HBV injection Oral treatment

5

Dai et al EASL 2018, SHC 2018, GHS 2018

The anti-HBV activity of RO7020531 in AAV-HBV infected mice relies on

functional adaptive immune response (B- and T- cells)

C57B/Bl6 (wild type mice) SCID (T- and B-cell deficient mice)

• Note: no change on HBeAg level was observed.

• Increasing dose levels of RO7020531 in SCID mice up to 300 mg/kg did not demonstrate anti-HBV activity.

• Plasma exposure of TLR7 agonist and innate immune responses (mRNA upregulation of interferon induced genes) in both

SCID and C57B/Bl6 mice were comparable6

Dai et al EASL 2018, SHC 2018, GHS 2018

The PD response of TLR7 signalling is observed in a dose-dependent

manner in RO7020531-treated AAV-HBV mice

* Mouse blood samples were collected at 6 hours post dose on day 28 for mRNA and cytokine tests.

TLR7 agonist RO7020531 Phase 1 umbrella study (NP39305)

8

• First-in-human study for

RO7020531

• 110 healthy volunteers

dosed in Part 1

• HBV patient part initiated

in 2018 (cohort 1 to 3 in

Nuc-suppressed patients

completed)

Part

1:

Co

mp

lete

d

150 mg QOD

Cohort 1

Cohort 2

Cohort 3

SAD in HV1 single dose

MAD in HV2 weeks of QOD dosing

Safety / PD Study in HBV Patients6 Weeks QOD treatment

6 Weeks follow-up

N total =110 (88:22)

Cohort 4

Cohort 2

Cohort 1

Cohort 3

Cohort 5

Cohort 6

Cohort 7

Cohort 8

170 mg

3 mg

10 mg

20 mg

40 mg

60 mg

100 mg

140 mg

Cohort 1

Cohort 2

Cohort 3

170 mg QOD

100 mg QOD

140 mg QOD

Part

2:

co

ho

rt 1

to

3

co

mp

lete

d

N per cohort =10 (8:2)

150 mg QOD

170 mg QOD

150 mg QOD Rx naive Cohort 4

Gane, E. et al. [poster]. EASL 2018; FRI-337

Gane, E. et al. SHC, 2018

Jin, Y. et al. APASL STC, 2019

RO7020531 has a favorable pharmacokinetics profile

• Dose proportional exposure across the

full RO7020531 dose range (3 mg–170 mg)

in SAD and MAD

• The active TLR7 agonist appears rapidly in

plasma with a median Tmax of ~0.5 hrs, is

cleared within 12 hours post dose (mean

half life ~ 3.0 hr)

• Stable RO7011785 exposure over a two

week dosing with no drug accumulation

• Following multiple QOD RO7020531

doses, there was no evidence for

RO7011785 accumulation, and PK

parameters on Day 1 and Day 13 in the

MAD were comparable

9

Concentration-time profiles for RO7011785 (active

agonist) following multiple QOD RO7020531 doses

0

500

1000

1500

2000

2500

3000

0 4 8 12 16 20 24

Time Post Dose (hr)

100 mg

140 mg

170 mg

0

500

1000

1500

2000

2500

3000

0 4 8 12 16 20 24

Time Post Dose (hr)

Day 1 Day 13

RO7011785 (ng/mL) RO7011785 (ng/mL)

Gane, E. et al. SHC, 2018; Jin, Y. et al. APASL STC, 2019

Pharmacodynamic activity first demonstrated at 100 mg (SAD)

• PD Evaluations: TLR activation markers in serum (IFN-α, IL-6, TNF-α, IL-10, IL-12p40, IP-10 and

neopterin) and expression of ISG15, OAS-1, MX1 and TLR7 mRNAs in whole blood

10

• Following a single 100 mg RO7020531 dose, 3/8 subjects exhibited increases in IFN-α indicative of TLR7

activation (Panel A)

• These changes in IFN-α were accompanied by increases above baseline for other markers of TLR7

activation including ISG-15 and OAS-1 shown as an example for Subject 1 (Panel B) following the

expected temporal pattern after the appearance of plasma RO7011785

Panel A Panel B

0.00

0.05

0.10

0.15

0.20

0.25

0

5

10

15

20

25

30

35

0 4 8 12 16 20 24

Time Post Dose (hr)

ISG15

OAS-1

Interferon

RO7011785

Fold

ch

an

ge

fro

m b

ase

line

(IS

G1

5 a

nd

OA

S-1

) Inte

rfero

n (p

g/m

L)

RO

701

17

85

(ug

/mL

)

0.00

0.05

0.10

0.15

0.20

0.25

0 4 8 12 16 20 24

Inte

rfe

ron (

pg

/mL

)

Time Post Dose (hr)

Subject 1

Subject 2

Subject 3

Baseline Value (dashed line)

Gane, E. et al. SHC, 2018; Jin, Y. et al. APASL STC, 2019

Higher RO7020531 doses yields more ‘responders’ and with a

greater amplitude of response (SAD cohorts)

With increasing single dose, the fraction of subjects exhibiting TLR7 PD activity and the magnitude of

response increased with a plateau of geometric mean response magnitude at 170 mg

11

Dose (mg)

ISG15 OAS1

Fraction

responding

Geometric mean fold

change (range)

Fraction

responding

Geometric mean fold

change (range)

Placebo 0/16 - 0/16 -

3 1/8 2.4 (2.4–2.4) 0/8 -

10 0/8 - 0/8 -

20 1/8 2.6 (2.6–2.6) 0/8 -

40 1/8 2.9 (2.9–2.9) 0/8 -

60 1/8 2.6 (2.6–2.6) 2/8 2.4 (2.0–2.9)

100 6/8 5.9 (2.3–29.3) 5/8 3.8 (1.9–6.9)

140 8/8 11.6 (2.3–48.0) 8/8 5.5 (2.2–18.1)

170 8/8 11.2 (2.5–132.2) 8/8 5.5 (2.0–19.0)

Gane, E. et al. SHC, 2018; Jin, Y. et al. APASL STC, 2019

PD activity maintained across multiple biomarkers with QOD

dosing of RO7020531

12

INF-α ISG15 mRNA

0

2

4

6

8

10

12

14

16

18

20

0 2 4 6 8 10 12 14

Inte

rfe

ron

(p

g/m

L)

Study Day

100 mg 140 mg 170 mg

0

20

40

60

80

100

120

0 2 4 6 8 10 12 14Fo

ld C

ha

ng

e F

rom

Ba

se

line

Study Day

100 mg 140 mg 170 mg

In the MAD cohorts, biomarker response was maintained over the 2 week QOD dosing

Jin, Y. et al. APASL STC, 2019

Summary

• RO7020531 is a novel double pro-drug of a TLR7 agonist.

• In the AAV-HBV mouse model, RO7020531 induced both innate PD responses and adaptive

immune responses which are important for therapeutic effects. The oral combination of the

CpAM and TLR7 agonist demonstrated robust suppression of both HBsAg and HBV DNA levels

and with the additional emergence of anti-HBs antibodies in several animals

• RO7020531 appears to be safe and tolerable with a predictable PK profile for the active TLR7

agonist in human.

• PD effects in human are observed with doses starting at 100 mg. At higher doses, more

responders and greater amplitude of response are observed.

• These promising preclinical results and Phase 1 clinical data provide encouragement to further

explore RO7020531’s therapeutic effect in chronic hepatitis B patients in combination with direct

acting antiviral agents.

13

14