SERUM HBV-RNA LEVELS DECLINE SIGNIFICANTLY IN CHRONIC HEPATITIS B PATIENTS DOSED WITH

THE NUCLEIC-ACID POLYMER REP 2139-Ca

Louis Jansen1,2, Andrew Vaillant3, Karel van Dort1, Femke Stelma1,2, Neeltje Kootstra1, Michel Bazinet3, Mamun Al-Mahtab4, Hendrik Reesink1,2

1 Experimental Immunology,2 Gastroenterology and Hepatology, Academic Medical Center, Amsterdam, Netherlands,

3 REPLICor Inc., Montreal, Canada,4 Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh

Disclosures

• Hendrik W. Reesink – Consulting / Research Support : Abbvie, BMS, Boehringer Ingelheim, Gilead, GSK, Janssen-Cilag, Merck/MSD, PRA-International, Regulus, Replicor, Roche, R-Pharm, Santaris.

• Andrew Vaillant, Michel Bazinet – Stockholders (shareholder) and Employees:

REPLICor Inc.

• The following people have nothing to disclose: Louis Jansen, Femke Stelma, Karel van Dort, Neeltje Kootstra, Mamun Al-Mahtab.

IntroductionChronic Hepatitis B Treatment

1. EASL. J Hepatol 2012.

Available therapies for patients with chronic hepatitis B1:• Peginterferon-α

+ Potential immune-mediated control of HBV- Significant side-effects

• Nucleos(t)ide analogues+ Potent viral supression- Limited off-therapy response

Need for new therapeutic approaches:

Enhance loss of HBeAg and HBsAg

Jansen EASL 2015 – 3

Hepatocyte

Virions

HBeAg

SVP(HBsAg)

Capsids

cccDNA

1. Zhang et al. J Med Virol. 20032. Bommel van et al. Hepatology. 20143. Jansen et al. AASLD 2013; EASL 2014

HypothesisSerum Hepatitis B RNA levels in NUC treatment?

? Serum HBV RNA1,2,3

Jansen EASL 2015 – 4

DNA

RNART

• NAPs have entry and post entry antiviral effects in HBV infection in vitro1

• The post-entry NAP effect appears to be linked to clearance of serum HBsAg2

Hypothesis:• NAPs prevent subviral particle (SVP) formation

REP 2139 = (A,5’MeC)20 PS-ON, fully 2’O-methylated

REP 2139-Ca = Calcium chelate complex of REP 2139(improves administration tolerability)

IntroductionNucleic Acid Polymers (NAPs) in Hepatitis B

1. Noordeen et al. Antimicrob Agents Chemother. 2013; 5291-5298 2. Noordeen et al. Antimicrob Agents Chemother. 2013; 5299-5306 Jansen EASL 2015 – 5

Serum Hepatitis B RNA levels in NAP treatment?

VirionsCapsids

cccDNA

Restoration ofimmune response?1,2

Elimination ofserum HBsAg

1. Wu et al. Hepatology. 20092. Boni et al. Gastroenterology. 2012

Hypothesis

Serum HBV RNA?

Jansen EASL 2015 – 6

Research Question

• Kinetics of serum HBV-RNA in patients dosed with nucleicacid polymer (NAP) REP 2139-Ca?

REP 102 protocol: Phase II proof of concept trialDr. Mamun Al-Mahtab, (Dhaka, Bangladesh)Dosing: 2011 – 2012, Follow-up ongoing

Jansen EASL 2015 – 7

Patient Cohort

• 12 Chronic Hepatitis B patients• HBeAg-positive• HBV DNA 105 – 108 copies / mL• Treatment naive• Metavir ≤ F3 (fibroscan)• ALT < 3 × ULN

Inclusion criteria REP 102 protocol

Jansen EASL 2015 – 8

Study dosing REP 102 protocol

Patient Cohort

REP 2139-Ca

Follow-up

20-24 Weeks

n = 12

REP 2139-Ca – 500mg qW 2 hour IV

Pegasys™ 180 ug SC qWand/or

Zadaxin™ (thymosin alpha-1) 1.6mg SC 2qW

Add-on*

21 Weeks (range 7-27)

13-26 Weeks

Entecavir

HBsAg non-responders (n = 3)

HBsAg responders (n = 9)

Jansen EASL 2015 – 9

*Add-on:

Patient Cohort

Baseline 20-24 Follow-up

• Serum (-20°C) sent to AMC for HBV RNA measurement• Compared to HBV DNA (Cobas), and HBsAg (Architect)

Time points of HBV RNA measurement

12

REP 2139-Ca

Jansen EASL 2015 – 10

REP 2139-Ca

Follow-up

20-24 Weeks

n = 12

Add-on*

21 Weeks (range 7-27)

13-26 Weeks

Entecavir

HBsAg non-responders (n = 3)

HBsAg responders (n = 9)

IM Add-on

Methods

• RNA isolation in plasma

• DNAse treatment

• Quantitative RT-PCR specific for HBV-RNA

1. Laras et al. Virology 2002.2. Jansen et al. AASLD 2013; EASL 2014.

Serum HBV RNA Quantification1,2

Jansen EASL 2015 – 11

4 6 8 10

2

4

6

8

10

HBV RNA (log10C/mL)

HB

sAg

(Log

10IU

/mL)

HBV

DNA

(Log

10C/

mL)

Results

HBV DNAr2 0.74, p < 0.001

HBsAgr2 0.33, p = 0.049

Baseline Serum HBV RNA Levels

Jansen EASL 2015 – 12

Baseline

RNA HBsAg-8

-6

-4

-2

0

2

Dec

line

from

bas

elin

e

Results

0 12 24

4

6

8

10

LLD

Weeks of REP 2139-Ca

HB

V R

NA

(log 1

0C/m

L)

n = 3

n = 9

Week 20-24 Decline

Serum HBV-RNA Decline During REP 2139-Ca Treatment

Jansen EASL 2015 – 13

NR

R

4

6

8

10

4

6

8

10

BL 20-24 FULLD LLD

IM12

HBV

DNA

HBV RNA

0

2

4

6

0

200

400

600

BL 20-24 FULLD

12 IM HB

sAg

anti-HBs

Results

4

6

8

10

4

6

8

10

BL 20-24 FULLD LLD

IM12

HBV

DNA

HBV RNA

0

2

4

6

0

200

400

600

BL 20-24 FULLD

12 IM HB

sAg

anti-HBs

At FU: 4/9 patientsHBsAg <0.05 IU/mL+ anti-HBs positive

Mean ± SEMHBV DNA, HBV RNA; log10 C/mLHBsAg; log10 IU/mLanti-HBs; U/L

Serum HBV-RNA in REP 2139-Ca Responders (n = 9)

Jansen EASL 2015 – 14

Results

0

2

4

6

0

200

400

600

BL 20-24 FULLD

12HB

sAg

anti-HBs

4

6

8

10

4

6

8

10

BL 20-24 FULLD LLD

12

HBV

DNA

HBV RNA

Mean ± SEMHBV DNA, HBV RNA; log10 C/mLHBsAg; log10 IU/mLanti-HBs; U/L

Serum HBV-RNA in REP 2139-Ca Non-Responders (n = 3)

ETV ETV

Jansen EASL 2015 – 15

Conclusions

• Treatment of CHB patients with REP 2139-Ca resulted in a pronounced decline of serum HBV-RNA in 9/12 of patients

• In 3/12 patients (non-responders) HBV-RNA levels were unaffected, both before and after treatment with entecavir.

• Related abstracts:

Jansen EASL 2015 – 16

Topic Abstract NumberPreclinical evaluation of NAPs in vivo P0542

HBV in vitro activity of NAPs P0556Clinical cytokine analysis (REP 102) P0659

HDV in vitro activity of NAPs LP26HBV / HDV clinical data (REP 301) LO2