Welcome to
Investor Day
June 29, 2016
Forward-looking statements
This information may contain projections and other forward‐looking statements regarding future events, including
regarding Dicerna’s technology platform, product candidates, preclinical and clinical pipeline and milestones,
regulatory objectives, market opportunities, and intellectual property. Such statements are predictions only and are
subject to risks and uncertainties that could cause actual events or results to differ materially. These risks and
uncertainties include, among others, the cost, timing and results of preclinical studies and clinical trials and other
development activities; the unpredictability of the duration and results of regulatory review of New Drug
Applications and Investigational NDAs; market acceptance for approved products and innovative therapeutic
treatments; competition; the possible impairment of, inability to obtain and costs of obtaining intellectual property
rights; and possible safety or efficacy concerns, general business, financial and accounting risks and litigation. More
information concerning Dicerna and such risks and uncertainties is available on its website and in its press releases,
and in its public filings with the U.S. Securities and Exchange Commission.
Dicerna is providing this information as of its date and does not undertake any obligation to update or revise it,
whether as a result of new information, future events or circumstances or otherwise. Additional information
concerning Dicerna and its business may be available in press releases or other public announcements and public
filings made after the date of this information.
2
Agenda and Speakers
Vision & Strategy
Overview of Opportunity Space
GalXC™ Platform Performance
Primary Hyperoxaluria: A New Approach
Focus on Intellectual Property
Overview: Chronic Liver Disease & Fibrosis
Douglas M. Fambrough, III, Ph.D.
President & Chief Executive Officer
Bob D. Brown, Ph.D.
Chief Scientific Officer, Senior Vice President
Bart Wise, Ph.D., J.D.
Vice President, Intellectual Property
Sanya Sukduang, J.D.
Partner
Finnegan, Henderson
3
Pankaj Bhargava, M.D.
Chief Medical Officer
Craig Langman, M.D.
Northwestern University
Rebecca Wells, M.D.
University of Pennsylvania
Executing Our Strategy
Douglas Fambrough, Ph.D.
President & Chief Executive Officer
Vision & Strategy
Douglas M. Fambrough, III, Ph.D.
President & Chief Executive Officer
GalXC™ Platform: Fully Enabled RNAi Drug Discovery Engine
Simple, potent, durable subcutaneous (SC) delivery to the liver
Powerful Capabilities
Expansive Opportunities
 Subcutaneous RNAi technology for liver targets
 Applicable across a wide variety of disease areas
 Potency supports gene silencing with a ≤1ml SC
injection volume in patients
 Long duration of action versus other modalities
 Very well tolerated, high therapeutic index
 Exquisite specificity to gene target
 Deep IP and freedom to operate
5
 Rare diseasesRare Diseases
 Chronic liver
diseases
Chronic
Liver Diseases
 Cardiovascular
diseases
Cardiovascular
Diseases
 Liver infectious
diseases
Liver Infectious
Diseases
 First-in-class and validated target programs
 Clear medical differentiation
Executing the Strategy
Harnessing RNAi’s natural, long-acting, catalytic mechanism for gene silencing
Development Strategy
• Retain substantial rights to high value
programs in focused, monogenic
indications such as rare diseases
• Seek strategic collaborators to develop
therapies for large patient populations
and select rare diseases
Reducing Risk
• Rare genetic diseases where gene silencing
generates a high probability of clinical success
 Primary hyperoxaluria type 1
 Undisclosed genetic metabolic disorders
• Validated targets for disorders with large patient
populations
 PCSK9
 HBV
• Seek strategic collaborations for complex disease
states
 Chronic liver diseases
 Cardiovascular diseases
6
GalXC Platform: A Different, Improved Approach
Unique, proprietary tetraloop construction offers distinct advantages
subcutaneous ● potent ● durable
• Multiple validations in monkeys
• 3 development candidates in 2016
• 3 or more strategic opportunities annually
• Rapid discovery: potent silencing in mice in 30 days
• Proprietary DsiRNA format drives strand selectivity
• Well established RNA modification chemistries
7
Tetraloop Advantages
• Provides enhanced stabilization
• Properly orients multiple ligands for
presentation to hepatocytes
• Simple ‘on-column’ oligo manufacturing
• Rapid generation time
How GalXC Delivers Conjugates to Liver Cells
8
Expansive Opportunities for RNAi Therapies in the Liver
Dicerna has qualified 29 potential disease targets to date
Rare Diseases
Chronic Liver
Diseases
Cardiovascular
Diseases
Liver Infectious
Diseases
HAO1
HMGB1
PCSK9
HBV
Primary
hyperoxaluria
type 1
Fibrotic liver
disease
Hypercholesterolemia
Chronic
hepatitis B
infection
10
qualified
targets
to date
8
qualified
targets
to date
7
qualified
targets
to date
4
qualified
targets
to date
The scope of the RNAi liver opportunity supports multiple successful entrants
9
Dicerna Today: Advancing GalXC Platform, Building a Broad Pipeline
Driven by capabilities. Drawn by opportunities.
Where We Are Today
3
New GalXC program launches in 2016
5
Projected clinical stage programs by
YE2019
2
Intravenous clinical programs headed to
clinical POC
$69M
10
Projected cash and ST investments
at end of Q2 2016
What You’ll See Today
• GalXC subcutaneous gene silencing for 12
different disease targets
‒ 6 examples in non-human primates
‒ 6 additional examples in rodents
‒ From idea to potent subcutaneous
silencing in rodents in 30 days
• GalXC-mediated efficacy for 6 different disease
targets
‒ 3 examples in rare disease models
‒ 3 examples in chronic liver disease models
The Platform Value of SC Oligo Delivery for the Liver
Historical precedent from humanized antibodies (diverse IP estates)
multiple products ● multiple winners
Value at acquisition/merger:
Genentech
$47B (2009)* Avastin®, Herceptin®, Rituxan®
MedImmune
$15.6B (2007)
Idec
$6.5B (2003)
Centocor
Medarex
Abgenix
Cambridge Antibody
11
$4.9B (1999)
$2.4B (2009)
$2.2B (2005)
$1.3B (2006)
Synagis®
Rituxan®
Remicade®
Yervoy® (Phase 3), pipeline
Vectabix® (Phase 3), pipeline
Humira® (royalty), pipeline
*value for portion not already owned by Roche
GalXC Development Pipeline – Dicerna Today
Product Candidate
Indication
Stage of Development
2017 Candidates 2016 Candidates
Research
12
DRNA 16.1: DCR-PHsc
Primary Hyperoxaluria
DRNA 16.2: DCR-undisclosed Orphan Genetic Disease
DRNA 16.3: DCR-PCSK9
DRNA 17.1
DRNA 17.2
DRNA 17.3
Cardiovascular Disease
Preclinical
Phase 1
More
Advanced
Studies
Pivotal
Trials
GalXC Development Pipeline Projected Growth – Dicerna 2019
Product Candidate
Indication
Stage of Development
2018 &
beyond
2017 Candidates 2016 Candidates
Research
13
DRNA 16.1: DCR-PHsc
Primary Hyperoxaluria
DRNA 16.2: undisclosed
Orphan Genetic Disease
DRNA 16.3: DCR-PCSK9
Cardiovascular Disease
DRNA 17.1
DRNA 17.2
DRNA 17.3
Dicerna has the capacity to launch 3 new
development programs each year
Preclinical
Phase 1
More
Advanced
Studies
Pivotal
Trials
GalXC Platform Performance
Bob D. Brown, Ph.D.
Chief Scientific Officer, Senior Vice President
Compelling and Consistent In Vivo Data Across Multiple GalXC Programs
Performance in rodents and non-human primates demonstrates GalXC platform potential
1.
2.
3.
4.
5.
15
Potent and long duration pharmacodynamic effects in vivo
Very high safety/tolerability in exaggerated pharmacology testing
Additional target genes silenced in non-human primates
Rapid generation of GalXC duplexes for target validation in polygenic diseases
Molecular description of the GalXC RNAi duplex platform
Targeting HAO1 to Treat Primary Hyperoxaluria Type 1
Silencing the HAO1 gene eliminates the key PH1 disease pathology
Primary hyperoxaluria type 1 (PH1) is rare genetic disease resulting in severe kidney damage caused by
excess production of oxalate in the liver
Assessments
Technical
Clinical
Commercial
Opportunity
16
Parameters
Investment Thesis
Validation
Genetic deletion of GO in mice reduces or eliminates hyperoxaluria
Biomarker
Urinary oxalate is easily measured, and is the cause of kidney damage that characterizes PH1
Glycolate is easily measured in urine or plasma as an on-target effect of HAO1 gene silencing
High Unmet Need
No highly efficacious therapeutic options available
Potentially fatal in the absence of a liver-kidney transplant
Patient Numbers
Estimated genetic incidence of 8 per million, in excess of 1,000 patients in US and EU registries
Patient Advocacy
Oxalosis and Hyperoxaluria Foundation (OHF)
GalXC HAO1 Screening in Monkeys, 3 mg/kg Single-Dose Duration of Action
HAO1 mRNA reduction out to 56 days after a single SC dose
GalXC HAO1 Screening, Single-dose Comparison
D P 4 6 2 1 P :D P 4 3 9 0 G
p re -d o s e )
m R N A
H A O 1
D P 3 6 9 2 P :D P 4 3 8 7 G
(re l to
R e m a in in g
P B S
1 0 0
7 5
D P 5 8 4 4 P :D P 5 8 4 3 G
D P 3 6 9 2 P :D P 5 4 5 2 G
5 0
%
2 5
1 0
0
0
1 0
2 0
3 0
D a y s
dose
17
4 0
5 0
6 0
One 3 mg/kg SC dose achieves 94%
maximum HAO1 mRNA silencing and
88% average silencing
• Single-injection 3 mg/kg dosing is
easily feasible in humans
• Historically, RNAi conjugates are
equally active or more active in
humans than they are in nonhuman primates
GalXC HAO1 Multi-Dose Pilot Study in Monkeys, Interim Results
Six month multi-dose regimen comparison, at Day 77 three to six doses were administered
One GalXC RNAi Duplex,
Four Different Dosing Regimens
1 0 0
Dose
Regimens:
(rel to pre-dose)
% HAO1 mRNA Remaining
Multi-Dose Regimens, HAO1 mRNA in NHP
Multi-dose Tx Regimens
2 mpk QMx4
7 5
2 mpk Q2Wx7
4 mpk QMx4
5 0
ongoing
4 mpk Q2Wx3;
2 mpk QMx2
2 5
Biopsy Days:
ongoing
1 0
Day -7
Day 28
Day 77
Day 133
0
0
7
1 4
2 1
2 8
3 5
4 2
4 9
5 6
6 3
7 0
7 7
8 4
9 1
9 8 1 0 5
D a y s
Equal or superior potency compared to
competitor SC inhibitor
18
Three out of four dosing regimens are
equivalent and efficacious after two months
93% maximum mRNA silencing (at Day 77)
GalXC HAO1 Multi-Dose Study in Monkeys
Multi-dose regimen comparison, plasma collected weekly, study ongoing
Plasma Glycolate Elevation in NHP after GalXC Treatment (to Day 63)
GalXC silencing of HAO1 mRNA (GO protein)
in normal non-human primates causes
elevation of plasma glycolate level, as
expected
P la s m a
G ly c o la te
(µ M )
6 0
5 0
4 0
3 0
Three out of four dosing regimens were
equally effective at elevating plasma
glycolate
2 0
1 0
0
-7
0
7
1 4
2 1
2 8
3 5
4 2
4 9
5 6
6 3
D a y
P B S
19
2
m p k Q M x 4
4
m p k
Q 2 W x 3 ; 2
2
m p k
Q 2 W x 7
4
m p k Q M x 4
m p k
Q M x 2
GalXC HAO1 PD Study in Monkeys Comparing mRNA, Protein & Glycolate
Multi-dose regimen comparison, plasma is collected weekly, study ongoing
GalXC HAO1 Dosing Regimen
PBS QMx4
4mg/kg Q2Wx3;
2 mg/kg QMx2
2mg/kg QMx4
2mg/kg Q2Wx7
4mg/kg QMx4
GO Protein
GAPDH
85%
98%
1 2 5
1 0 0
1 0 0
2
=
0 .9 5
7 5
P la s m a
P r o te in
P B S )
(R e l to
5 0
R
P r o te in
5 0
2 5
2 5
% GO Protein vs Glycolate, Day 77
1 0 0
m R N A
7 5
Avg Protein
Reduction
92%
% HAO1 mRNA vs GO Protein, Day 77
1 2 5
%
H A O 1
R e m a in in g
% HAO1 mRNA and GO Protein, Day 77
97%
G ly c o la te
0%
8 0
6 0
4 0
2 0
1 0
0
2
4
2 5
5 0
7 5
1 0 0
0
2 5
5 0
7 5
M
Q
W
k
p
k
4
m
p
m
p
m
2
2
Q
x
k
p
m
4
0
0
x
7
x
;
3
x
2
W
Q
Q
k
p
m
2
20
M
k
2
Q
P
M
B
x
S
4
0
m R N A
P r o te in
1 0 0
1 2 5
Hepatic Effects of Alpha-1 Antitrypsin (AAT)
Common AAT mutant alleles result in misfolded protein accumulation in hepatocytes
AAT deficiency is a hereditary disorder resulting in an increased incidence of chronic liver disease
leading to cirrhosis and hepatocellular carcinoma in affected individuals.
Assessments
Technical
Parameters
Therapeutic
Hypothesis
The inherited, Z form of Alpha-1 protein polymerizes and accumulates in liver causing
hepatotoxicity. Preventing its synthesis may prevent liver disease
Biomarker
Alpha-1 mutant Z polymerized protein in serum (emerging concept)
High Unmet Need
There are no approved therapies for the treatment of AAT-deficient patients with liver
disease, the primary metabolic liver disease leading to pediatric liver transplant
Development
Evolving endpoints and path for the treatment of liver disease
Natural history studies underway to provide further knowledge in pediatric and adult
patients
Patient Numbers
~20,000 EU/US adult patients with liver disease
~10,000 EU/US pediatric patients with liver disease
Market/Market
Leverage
WW augmentation market >$500MM; recent DD growth units & revenue
Weekly IV infusion used by some centers
Patient Advocacy
Alpha-1 Foundation represents both lung and liver disease patients
Clinical
Commercial
Opportunity
21
Investment Thesis
GalXC AAT Screening in NHP: 3 mg/kg Single Dose Duration of Action
One SC dose of three different GalXC duplexes in monkeys through Day 75, study ongoing
Single-Dose Duration of Action, GalXC AAT Silencing
3 mg/kg, one dose,
test articles
DP4620:DP5081G
DP4620:DP5080G
7 5
DP4620:DP5082G
5 0
%
N H P
S e ru m
A A T
1 0 0
2 5
• Long-term comparison of different
GalXC chemistries on one AAT
sequence, each administered to
monkeys SC at 3 mg/kg, one dose
• All GalXC duplexes yielded
sustained protein reductions
• The most efficacious GalXC-AAT
duplex achieved maximum protein
reduction over two months after a
single dose (76% avg.)
0
0
1 4
dose
22
2 8
4 2
5 6
7 0
8 4
9 8
D a y s
89% maximum serum AAT protein reduction after a single dose (Day 68)
Equipotent and superior duration compared to competitor SC inhibitor
Duration of GalXC Action: Long-Term Serum PiZ Protein Reduction in Mice
SC dosing at 0.25, 0.5, 1 mg/kg QWx4 vs. 1 mg/kg Q5Wx2, PiZ GEMM (Teckman Lab)
(rel to time-matched PBS)
% PiZ Serum Protein
GalXC Treatment Regimen Comparison for SERPINA1 (Alpha-1 Antitrypsin)
P B S
1 0 0
0 .2 5
0 .5
m p k
m p k
2 5
1
m p k
Q W x 4
1
m p k
Q 5 W x 2
0
0
7
1 4
2 1
2 8
3 5
4 2
4 9
SC dosing
regimens
23
Q W x 4
5 0
1 0
•
•
•
•
Q W x 4
5 6
6 3
7 0
7 7
8 4
9 1
Days
96% maximum PiZ protein reduction after Q5Wx2 and 99% after QWx4, respectively
Repeated dosing as low as 0.25 mg/kg was efficacious
Two 1 mg/kg doses five weeks apart was as effective as four weekly doses at 1 mg/kg
The GalXC platform enables flexible, infrequent dosing regimens
GalXC Undisclosed Target #1 (UDT #1): Indications & Clinical
Characteristics
Related family of rare genetic diseases resulting in life-threatening conditions and multi-organ
complications. Various specific disease types differ in their severity and genetic cause; however,
we believe there may be a target common to all disease types.
Assessments
Technical
Commercial
Opportunity
24
Parameters
Investment Thesis
Validation
Target common to multiple rare diseases in the same family each with differing genetic causes
Genetic deletion of target in mice reduces or eliminates major pathology of disease
Biomarker
Single plasma biomarker associated with the disease family
High Unmet Need
No highly efficacious therapeutic options available
Potentially fatal in the absence of major medical intervention
Patient Numbers
Estimated genetic incidence in excess of 10 per million
Competition
Best-in-class opportunity
Challenging target for small molecules and antibodies
Patient Advocacy
Active patient groups and growing disease awareness
GalXC UDT #1: Lead Screening in Mice, Single Dose IC50 ≈0.1 to 0.3 mg/kg
Single-dose, dose response, 72 hours, murine HDI model, multiple chemistry alternatives
% UDT #1 mRNA Remaining
Undisclosed Target #1 GalXC Screen, Dose Response
Screening for GalXC optimization in vivo
1 0 0
7 5
5 0
2 5
1 0
0
P B S
0 .1
0 .3
1
3
GalXC #1.5
0 .1
0 .3
1
GalXC #1.6
0 .1
0 .3
1
GalXC #1.7
0 .3
1
3
GalXC #1.8
0 .3
1
3
Dose
mg/kg
GalXC #1.9
Different GalXC Duplexes and Dose
Optimization of GalXC hits in mice routinely produces
duplexes with IC50 of 0.1 to 0.5 mg/kg after a single dose
25
GalXC UDT #1 Screening in NHP: 3 mg/kg Single Dose Duration of Action
GalXC UDT #1 mRNA reduction comparison out to 56 days after a single 3 mg/kg SC dose
p re -d o s e )
m R N A
# 1
7 5
5 0
In mouse models of Undisclosed Target #1
diseases, therapeutic benefit is proportional
with mRNA and protein silencing
• Two test articles produced >75% avg. and 88%
P B S
G a lX C
1 .1
G a lX C
1 .2
G a lX C
1 .3
G a lX C
1 .4
maximum mRNA silencing at Day 14 after a
single 3 mg/kg dose
• Almost two months (Day 56) after one 3
mg/kg dose, GalXC 1.2 still maintained 63%
average and up to 80% max target silencing
2 5
%
U D T
1 0 0
(re l to
R e m a in in g
Single-Dose Duration Against UDT #1 Target in NHP
0
0
1 0
2 0
3 0
4 0
5 0
6 0
D a y s
dose
GalXC leads are potent and have a long duration of action in monkeys
26
GalXC UDT #1: Multi-Dose Regimens in NHP
GalXC UDT #1 mRNA reduction normalized to pre-dose biopsy
One GalXC RNAi Duplex,
Four different Dosing Regimens
Multi-Dose Regimens, UDT #1 mRNA in NHP
Dose
Regimens:
p re -d o s e )
m R N A
# 1
4 mpk QMx4
4 mpk Q2Wx3;
2 mpk QMx2
5 0
Biopsy Days:
Pre-dose
2 5
ongoing
%
U D T
2 mpk Q2Wx7
7 5
1 0
0
0
7
1 4
2 1
2 8
3 5
4 2
4 9
5 6
D a y s
27
Multi-dose Tx Regimens
2 mpk QMx4
(re l to
R e m a in in g
1 0 0
6 3
7 0
7 7
8 4
9 1
9 8 1 0 5
Day 29
Day 61
Day 104
All groups achieved ≥85% mRNA silencing by two
months (ongoing)
95% maximum mRNA silencing (Day 61, ongoing)
Compelling and Consistent In Vivo Data Across Multiple GalXC Programs
Performance in rodents and non-human primates demonstrates GalXC platform potential
1.
2.
3.
4.
5.
28
Potent and long duration pharmacodynamic effects in vivo
Very high safety/tolerability in exaggerated pharmacology testing
Additional target genes silenced in non-human primates
Rapid generation of GalXC duplexes for target validation in polygenic diseases
Molecular description of the GalXC RNAi duplex platform
GalXC UDT #1: Multi-Dose Exaggerated Pharmacology Study
High dose up to 100 mg/kg GalXC QWx6 followed by CBC and histopathology, CD-1 mice
CD-1 Wild-type mice
GalXC UDT #1 duplex administered Q1Wx6, 1, 3, 10, 30 or 100 mg/kg
week 1
week 2
week 3
week 4
week 5
week 6
Take down 24 hr
after the last dose
Plasma Marker
(mmol/L)
(% of starting weight )
In the
normal range
B o d y
5
W e ig h t)
o f S ta r tin g
W e ig h t C h a n g e
1 1 5
(%
10
• Extended silencing of UDT #1
Body Weight Change
1 1 0
1 0 5
1 0 0
P B S
9 5
1
m
3
m
g /k g
g /k g
1 0
m g /k g
3 0
m g /k g
9 0
1 0 0
m g /k g
8 5
0
0
PBS
29
1
3
10
30
100
1
2
W e e k s
3
p o s t d o s e
4
5
6
mRNA in mice produced no
clinical observations
• Behavior and body weight were
normal
• Full histopathology was
performed on liver, kidney and
skin in the injection region
(between the shoulder blades)
• Plasma marker levels were in the
normal range for these animals
GalXC UDT #1: NOAEL is >100 mg/kg Administered Weekly
QWx6 SC dosing of 1, 3, 10, 30 and 100 mg/kg of GalXC UDT #1 duplex in CD-1 mice
liv e r )
(n g /g
P B S )
7 5
to
5 0
1 0 0 0 0
1 0 0 0
G a lX C
2 5
1 0 0
B L O Q
0
P B S
1 .0
m g /k g
3 .0
m g /k g
1 0
m g /k g
3 0
m g /k g
1 0 0
P B S
m g /k g
ALT
1 .0
m g /k g
AST
80
150
A S T ( U /L )
A L T ( U /L )
60
100
40
50
20
0
0
PBS
30
1 .0 m p k
3 .0 m p k
10m pk
30m pk
100m pk
PBS
1 .0 m p k
3 .0 m p k
10m pk
3 .0
A L K A L IN E P H O S P H A T A S E ( U /L )
m R N A
1 0 0
%
U T # 1
GalXC UDT #1 Liver Exposure
1 0 0 0 0 0
( R e la tiv e
R e m a in in g
UDT #1 mRNA KD
30m pk
100m pk
m g /k g
1 0
m g /k g
3 0
m g /k g
1 0 0
m g /k g
ALP
200
150
100
50
0
PBS
1 .0 m p k
3 .0 m p k
10m pk
30m pk
• Body weight gain among groups was identical, there were no clinical observations
• Liver, kidney and injection site histopathology shows no adverse events, even at 100 mg/kg
100m pk
GalXC UDT #1: 100 mg/kg QWx6 Produced No SAEs in Mice
From the histopathology report
Tissue
Histopathology
Granulomatous foci
Extramedullary hematopoiesis
Inflammatory foci
Degeneration
Necrosis
Liver
Autolysis
Mineralization
Neoplasia
Hyperplasia
Vasculitis
Atrophy
Hemorrhage
Dilation, tubular
Inflammation, renal pelvis
Inflammation, cortex
Tubule loss, cortex
Kidney
Autolysis
Mineralization
Dilation, vascular
Hyperplasia
Vasculitis
Atrophy
Epidermal crust/ulceration
Edema
Inflammation, dermal or subcutaneo
Degeneration
Necrosis
Injection Site
Autolysis
Mineralization
Neoplasia
Hyperplasia
Vasculitis
31
Atrophy
PBS
1 mg/kg (N=5)
3 mg/kg (N=5)
10 mg/kg (N=5)
30 mg/kg (N=5)
100 mg/kg (N=5)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (mil)
0
0
0
0
0
0
0
0
0
0
1 (min)
1 (mil)
0
0
0
0
0
0
0
1 (mil)
0
1 (mil)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
3 (min)
0
0
1 (min)
0
0
0
0
0
0
0
0
1 (min)
0
0
0
0
0
0
0
0
1 (min)
0
2 (min)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (min)
0
1 (min)
0
0
0
0
0
0
0
0
2 (min, mil)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
1 (mil)
0
1 (mil)
0
0
0
0
0
0
0
0
min: minimal
1 (mil)
1 (mil)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
2 (min, mil)
0
4 (1 min, 3 mil)
0
0
0
0
0
1 (min)
0
0
mil: mild
• No SAEs were observed
• Observations were all
“minimal” to “mild”
• Dermal observations were
associated with the actual
skin puncture site and
needle track
• Kidney observations were
sporadic and not
associated with the test
article
• Liver observations were
not adverse and not clearly
associated with test article
No Cytokine Induction After SC Administration of GalXC to Monkeys
Pre-dose vs. 3 hours post one SC dose from 1 to 5 mg/kg, n=4/group; pilot study
IF N - γ
IFN-gamma
IL - 1 β
IL-1beta
100
C o n c e n t r a t io n ( p g /m L )
250
IL-8
IL -8
100
4 ,0 0 0
200
3 ,0 0 0
150
50
50
2 ,0 0 0
100
1 ,0 0 0
50
0
0
0
3
0
1
3
0
3
3
0
5
3
0
3
3
0
0
3
3
0
1
3
0
3
3
0
5
3
0
3
3
0
0
3
3
0
1
3
0
3
3
0
5
3
0
3
3
0
3
3
0
1
3
0
3
3
0
5
3
0
3
3
h r s
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4 6 2 0 P :
D P 4 6 2 0 P :
D P 4 6 2 0 P :
D P 5081G
D P 5080G
D P 5082G
D P 5081G
D P 5080G
D P 5082G
D P 5081G
D P 5080G
D P 5082G
D P 5 0 8 1 G
D P 5 0 8 0 G
D P 5 0 8 2 G
M IP -1 α
MIP-1alpha
M IP -1 β
MIP-1beta
100
1 ,0 0 0
p o s t-d o s e
d o s e
3
D P 4620P:
M
C P -1
MCP-1
C o n c e n t r a t io n ( p g /m L )
IL
-6
IL-6
(m g /k g )
T N F -α
TNF-alpha
150
1 0 0
100
50
500
5 0
50
0
0
0
3
1
32
0
3
3
0
3
5
0
3
3
0
3
3
0
0
3
1
0
3
3
0
3
5
0
3
3
0
3
3
0
0
3
1
0
3
3
0
3
5
0
3
3
0
3
3
0
3
1
0
3
3
0
3
5
0
3
3
0
3
3
h r s
p o s t-d o s e
d o s e
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4620P:
D P 4 6 2 0 P :
D P 4 6 2 0 P :
D P 4 6 2 0 P :
D P 5081G
D P 5080G
D P 5082G
D P 5081G
D P 5080G
D P 5082G
D P 5081G
D P 5080G
D P 5082G
D P 5 0 8 1 G
D P 5 0 8 0 G
D P 5 0 8 2 G
(m g /k g )
Compelling and Consistent In Vivo Data Across Multiple GalXC Programs
Performance in rodents and non-human primates demonstrates GalXC platform potential
1.
2.
3.
4.
5.
33
Potent and long duration pharmacodynamic effects in vivo
Very high safety/tolerability in exaggerated pharmacology testing
Additional target genes silenced in non-human primates
Rapid generation of GalXC duplexes for target validation in polygenic diseases
Molecular description of the GalXC RNAi duplex platform
GalXC Hits Against Three Additional Undisclosed Targets in NHP
The platform enables rapid discovery and optimization
Multiple Targets, Single Dose, 3 mg/kg, Day 21
• GalXC UDT #2 and GalXC UDT #3 were POC
T a rg e t m R N A
R e m a in in g
1 0 0
7 5
≥75% mRNA
reduction
5 0
>95% mRNA
reduction
%
2 5
duplexes to test early GalXC function in NHP
• UDT #2 and UDT #3 resulted in
approximately 75% target mRNA silencing
• Three GalXC duplexes against UDT #4 were
designed using advanced GalXC concepts
• Each GalXC UDT #4 duplex yielded >95%
mRNA silencing after a single 3 mg/kg dose
1 0
.3
U
D
T
#
4
.2
T
D
U
C
D
o
T
#
#
4
tr
n
T
D
U
U
34
4
.1
l
#
o
3
l
o
tr
n
o
C
U
C
o
D
n
T
tr
#
o
2
l
0
GalXC hits against multiple therapeutic gene targets in monkeys are
advancing rapidly through preclinical development
Compelling and Consistent In Vivo Data Across Multiple GalXC Programs
Performance in rodents and non-human primates demonstrates GalXC platform potential
1.
2.
3.
4.
5.
35
Potent and long duration pharmacodynamic effects in vivo
Very high safety/tolerability in exaggerated pharmacology testing
Additional target genes silenced in non-human primates
Rapid generation of GalXC duplexes for target validation in polygenic diseases
Molecular description of the GalXC RNAi duplex platform
Rapid Generation of GalXC Duplexes to Probe Complex Disease Genetics
GalXC design in silico to in vivo testing in mice a single step, 30 days from gene to results
In silico RNAi sequence selection
+
Application of GalXC design “rules”
Nick
1 mg scale, 12 to 36 GalXC duplexes
In vivo screening, one SC dose
ED50 ≤1 mg/kg GalXC duplexes
Rapid GalXC Application to In Vivo Disease Models
36
• Sequence selection plus chemical
modification algorithms enable one-step
generation of mouse-specific, highly potent
GalXC duplexes
• High-throughput manufacture of GalXC
duplexes enables rapid turnaround time
It is now possible to mine complex,
polygenic indications in mouse models of
disease to identify and confirm the gene
targets with true therapeutic potential
37
N o v e l C L D
G a lX C
S c re e n , S e q u e n c e s
v s . M o d ific a t io n s
Independent sequences + modification patterns = confirmation of GalXC design rules
D P 8 1 0 8 G
D P 8 1 5 9 P :
D P 8 1 0 6 G
D P 8 1 5 8 P :
D P 8 1 0 5 G
S M 1 0 4 7 /A S M 1 0 5 6
D P 8 1 5 7 P :
D P 8 1 0 2 G
Sequence 4
D P 8 1 5 7 P :
D P 8 1 0 2 G
D P 8 1 5 6 P :
D P 8 1 0 0 G
D P 8 1 5 5 P :
D P 8 0 9 8 G
D P 8 1 5 4 P :
D P 8 0 9 7 G
S M 9 8 8 /A S M 1 0 2 1
D P 8 1 5 3 P :
D P 8 0 9 4 G
Sequence 3
D P 8 1 5 3 P :
D P 8 0 9 4 G
D P 8 1 5 2 P :
D P 8 0 9 2 G
D P 8 1 5 1 P :
D P 8 0 9 0 G
S M 9 8 8 /A S M 1 0 2 2
D P 8 1 5 0 P :
D P 8 0 8 9 G
D P 8 1 4 9 P :
7 5
D P 8 0 8 6 G
Sequence 2
D P 8 1 4 9 P :
D P 8 0 8 6 G
D P 8 1 4 8 P :
D P 8 1 4 0 G
D P 8 1 4 7 P :
S M 1 1 4 4 /A S M 1 0 2 2
:D P 8 1 3 9 G
D P 8 1 4 6 P
D P 8 1 3 8 G
D P 8 1 4 5 P :
D P 8 1 3 7 G
# 5
Sequence 1
D P 8 1 4 5 P :
D P 8 1 3 7 G
D P 8 1 4 4 P :
D P 8 0 7 6 G
D P 8 1 4 3 P :
R e m a in in g , U D T
P a tte rn s :
D P 8 0 7 4 G
D P 8 1 4 2 P :
D P 8 0 7 3 G
D P 8 1 4 1 P :
D P 8 0 7 0 G
m R N A
M o d
D P 8 1 4 1 P :
D P 8 0 7 0 G
D P 8 1 6 0 P :
P B S
%
GalXC UDT #5: Applying In Silico Design Rules for GalXC Activity in Mice
in One Month
3 mg/kg, single SC dose, 72 hour harvest – 87% target silencing
Five Algorithm-Selected Sequences x Five Chemical Modification Patterns, All Screened In Vivo
S M 1 1 4 4 /A S M 1 0 2 2
Sequence 5
1 0 0
87% silencing
ED50 = 0.5 mg/kg
5 0
2 5
1 2 .5
0
GalXC UDT #6: 97% mRNA Silencing In Vivo in 30 Days with Murine Model
Single dose, SC 5 mg/kg, 96 hour harvest, C56Bl6 mice
Twelve Algorithm-Selected Sequences x One Modification Pattern
target gene were selected in silico
• One high in vivo potency chemical
1 0 0
modification pattern was applied to
each test sequence
7 5
(re l to
P B S )
• Half of the resulting GalXC duplexes
gave an average of 90 to 95% mRNA
reduction
5 0
90%
silencing
%
N o v e l T a rg e t m R N A
• Twelve test sequences against a novel
• The maximum mRNA silencing achieved
2 5
was 97% after a single dose
1 0
38
2
6
#
T
U
D
T
D
U
D
U
U
Chemistry SM1047/ASM1056
.1
1
6
#
T
T
D
T
U
D
U
D
T
T
U
D
T
U
D
T
U
D
T
U
D
T
U
D
T
D
U
Test GalXC Sequences
#
6
.1
0
.1
.9
6
#
6
#
#
#
6
.8
.7
.6
6
.5
#
6
#
#
6
.4
.3
6
.2
#
P
#
6
6
B
S
.1
0
This in silico-to-in vivo capability
is a result of testing thousands of
chemical modification patterns
and thousands of sequences
Mining the CLD Target Space to Identify Validated Therapeutic Targets
Focus on those where knockdown in hepatocytes improves outcomes in CLD preclinical models
Possible gene targets
ACC1/2
ASK1
CASP1
CB1
COL1A1
CTGF
CTNNB1
CYP27A1
CYP7A1
DGAT2
FOXF1
HIC5
HMGB1
HSP47
LOX
LOXL2
MYC
OPN
PLIN5
PNPLA3
SERPINA1
SHH
SMAD 2/3/7
SNAI1
SRBP1C
TGFβ
TM6SF2
Others
Sound
therapeutic
hypothesis
Expressed in
hepatocytes
Preclinical data
from genetic
models
Proof of concept
phenotypic benefit in
accepted, predictive
preclinical models to
support clinical
development lead
candidates
Target Filtering steps
Parallel workstreams:
Input from partners and KOLs
Medical and commercial analysis
Target to indication mapping (e.g. NASH, NAFLD, PSC)
GalXC hit identification, medicinal chemistry and IP
Complex polygenic disease spaces can be “mined” to validate therapeutic targets
39
Compelling and Consistent In Vivo Data Across Multiple GalXC Programs
Performance in rodents and non-human primates demonstrates GalXC platform potential
1.
2.
3.
4.
5.
40
Potent and long duration pharmacodynamic effects in vivo
Very high safety/tolerability in exaggerated pharmacology testing
Additional target genes silenced in non-human primates
Rapid generation of GalXC duplexes for target validation in polygenic diseases
Molecular description of the GalXC RNAi duplex platform
GalXC Features Deliver a Fully Enabled RNAi Platform
Passenger (sense strand) Design Rules
Guide (antisense strand) Design Rules
Constant sequence
Nicked
Constant sequence
• Guide/Passenger duplex sequences are algorithm-selected
• Potent sequences are placed into the proprietary GalXC tetraloop context
The stem/tetraloop sequence and chemistry are constant
• Chemical modification of each strand is optimized by sequence
Natural and industry-standard chemical modifications are utilized
• The proprietary GalXC RNAi trigger configuration places the Guide strand and the GalNAc sugars
into an ideal context for delivery to hepatocytes and RNAi activity
-
41
GalXC Duplexes: Natural and/or FDA-approved Nucleotide Components
Toxicology risk is low; manufacturing and CMC are well established
Example of Typical GalXC RNAi duplex
metabolism – degradation by nuclease, phosphatase,
esterase and peptidase activities, not P450-mediated
Guide Strand Components: 22 nucleotides
42
Passenger Strand Components: 36 nucleotides
11x 2′-OMe ribose (natural)
23x 2′-OMe ribose (natural)
11x 2′-F deoxyribose (FDA approved)
9x
2′-F deoxyribose (FDA approved)
5x
phosphorothioate (FDA approved)
1x
phosphorothioate (FDA approved)
1x
novel mononucleotide
4x
GalNAc-nucleotide (becomes natural
GalNAc + short linker + natural RNA)
Synthesis of GalXC Passenger Strands With GalNAc for Liver Targeting
This is independent of the GalXC RNAi configuration
Synthesis
5’-
DMTrO
-3’-
• GalNAc-phosphoramidites are
compatible with standard synthesis
and purification processes
• GalNAc-phosphoramidites are
simple to synthesize and scale up
DMTrO
B
or
O R
P
CEO
N(iPr)2
3
DMTrO
HO
B
B
O O
P
CEO
N(iPr)2
p. 1402, Zatsepin and Oretskaya
Chemistry and Biodiversity 2004,1,1401
GalNAc Phosphoroamidite
O
O
C O
R
Detritylation
DCA
R
C O
2) Capping
Ac2O
2
1
DMTrO
B
O
HO
B
CEO
=
CPG
B = Nucleobases:
Uracil,
N4-Acetylcytosine,
N6-Benzoyladenine,
N2-Isobutyroylguanine
R = OMe, F, or any 2'-modification
O O
P
CEO
O
R
DCA = Dichloroacetic acid
+
R
B
O
H 3C
C O
B
O
O
O
B
O
C O
O
R
4
C O
7
DMTr = Dimethoxytrityl
CE = Cyanoehtyl
O
P
O
O
4) Detritylation
DCA
R
DMTrO
B
O
O O
P
CEO
O
R
3) Oxidation
I2/H2O/Pyridine
B
O
O
C O
6
43
AcHN
O
O
OAc
OAc
1) Coupling
Tetrazole
O
O
OAc
O
“The most evident way to
incorporate carbohydrate residues
[e.g. GalNAc] into oligonucleotides is
to synthesize their
phosphoramidite derivatives…”
B
O
O
R
C O
5
R
GalXC RNAi Discovery and Application Timeline
2008
License of DsiRNA platform and invention and patent application filings covering the
GalXC RNAi structures
2013
Reduction to practice of GalXC structures
Q3 2013 – expanded internal chemistry efforts
2014
2015
EC50 >10 mg/kg
Q2 2014 – initial SC GalXC duplex in vivo EC50 of >10 mg/kg
Q4 2014 – announcement of SC potency of ~2mg/kg
All future liver disease programs to be GalXC-based instead of lipid nanoparticles,
oncology remains LNP-based
Q2 2015 – translation of activity from rodents to non-human primates
Q3 2015 – announcement of SC potency of ~0.6 mg/kg
GalXC POC in monkeys for multiple therapeutic targets
2016
44
Q1 2016 – Last chemical optimizations, key monkey data for 3 targets
Q2 - Q4 2016 – Clinical Lead declarations for three DRNA programs
EC50 ~2 mg/kg
EC50 <1.0 mg/kg
EC50 ≤0.3 mg/kg
EC50 = 50% reduction of mRNA levels in mice
GalXC Platform Summary
Validation of GalXC in rodents and non-human primates demonstrates therapeutic potential
•
•
•
•
•
45
In vivo pharmacodynamic data in mice and monkeys
- GalXC RNAi duplexes are potent and have a long duration of action in rodents and primates,
enabling flexible, infrequent dosing regimens that are predictive of clinical utility
Multiple GalXC programs
In vivo mRNA silencing was demonstrated with more than a dozen preclinical gene targets
High therapeutic index
Preclinical safely/tolerability signals in exaggerated pharmacology studies
30-day target mRNA silencing in vivo
GalXC design to in vivo testing is very rapid, enabling target validation in complex diseases
GalXC molecular details
Proprietary GalXC features and well-established industry chemistries deliver a fully enabled
RNAi platform
GalXC hits are advancing rapidly through preclinical development
Focus on Intellectual Property
Bart Wise, Ph.D., J.D.
Vice President, Intellectual Property
Sanya Sukduang
Partner, Finnegan, Henderson
• Finnegan, Henderson
-
Global IP Firm of the Year
(Managing Intellectual Property Global IP Awards, 2016)
-
U.S. Specialty IP Firm of the Year
(Managing Intellectual Property North American IP Awards, 2016)
-
U.S. IP Firm of the Decade
(Managing Intellectual Property North American IP Awards, 2015)
• Sanya Sukduang practices patent litigation in the pharmaceutical and biotechnology space before the
Federal Circuit and U.S. District Courts and post-grant work before the PTAB
• Represented companies such as Eli Lilly, AbbVie, Aventis, Allergan, GlaxoSmithKline, Valeant, Forest
Labs
• Awards
- Eli Lilly’s “Quality Advocate Award” for work on successfully defending Lilly’s Cymbalta product
- LMG Life Sciences “IP: Life Science Star” 2012-2015
- NAPABA 2013 Best Lawyers Under 40
47
Finnegan, Henderson, Farabow, Garrett & Dunner, LLP
Dicerna Investor Day
June 29, 2016
Sanya Sukduang
48
Disclaimer
These materials have been prepared solely for educational and entertainment purposes to
contribute to the understanding of U.S. and European intellectual property law. These materials
reflect only the personal views of the authors and are not individualized legal or investor advice. It is
understood that each case is fact specific, and that the appropriate solution in any case will vary.
Therefore, these materials may or may not be relevant to any particular situation. Thus, the authors,
Finnegan, Henderson, Farabow, Garrett & Dunner, LLP (including Finnegan Europe LLP, and Fei
Han Foreign Legal Affairs Law Firm) cannot be bound either philosophically or as representatives of
their various present and future clients to the comments expressed in these materials. The
presentation of these materials does not establish any form of attorney-client relationship with these
authors. While every attempt was made to ensure that these materials are accurate, errors or
omissions may be contained therein, for which any liability is disclaimed.
49
49
49
Speaker Information
Sanya Sukduang concentrates on patent litigation before
federal district courts and the U.S. Court of Appeals for the
Federal Circuit, primarily in the areas of biotechnology,
pharmaceuticals, biologics, and medical devices. He has
conducted all aspects of pre-trial, trial, and post-trial
proceedings. Mr. Sukduang has extensive experience in
cases arising from the filing of Abbreviated New Drug
Applications (ANDA).
Sanya Sukduang
Finnegan, Henderson,
Farabow, Garrett & Dunner, LLP
901 New York Avenue, NW
Washington, DC 20001-4413
+1 202 408 4377
+1 202 408 4400 Fax
sanya.sukduang@finnegan.com
50
50
50
The Purpose and Scope of Patent Protection
• “[T]he patent system represents a carefully crafted bargain that encourages both
the creation and the public disclosure of new and useful advances in technology,
in return for an exclusive monopoly for a limited period of time.”
– Pfaff v. Wells Electronics, Inc., 525 U.S. 55, 63 (1998)
• “Claims define the subject matter . . . . Their principal function, therefore, is to
provide notice of the boundaries of the right to exclude and to define limits. . . .
Claims define and circumscribe.”
– Ariad Pharm., Inc. v. Eli Lilly & Co., 598 F.3d 1336, 1346 (Fed. Cir. 2010)
51
51
51
Patent Laws Protect Against Inventors Claiming More
Than What They Invented
• “[T]he purpose of the written description requirement is to ‘ensure that the
scope of the right to exclude, as set forth in the claims, does not overreach
the scope of the inventor's contribution to the field of art as described in
the patent specification.’”
– Ariad Pharm., Inc. v. Eli Lilly & Co., 598 F.3d 1336, 1353-54 (Fed. Cir. 2010)
quoting Univ. of Rochester v. G.D. Searle & Co., Inc., 375 F.3d 1303 (Fed.
Cir. 2004)
• Patent owners are not permitted to exaggerate the scope of a patent or
give the false impression that they are the exclusive source of a product or
technology.
– Zenith Elecs. Corp. v. Exzec, Inc., 182 F.3d 1340
(Fed. Cir. 1999)
52
52
52
Inventors Build and Improve Upon What Was Known
• “[T]echnologic advance, like scientific advance, usually proceeds in small,
incremental steps, building on what has gone before, building on one's own work
and the work of others. The steps, accumulating, may eventually produce the
next generation of technologic progress.”
– Hilton Davis Chemical Co. v. Warner-Jenkinson Co., Inc., 62 F.3d 1512, 1533 (Fed.
Cir. 1995)
• “[P]atent claims should not prevent the use of the basic building blocks of
technology . . . .”
– Ariosa Diagnostics, Inc. v. Sequenom, Inc., 788 F.3d 1371, 1379 (Fed. Cir. 2015)
53
53
53
Focus on Intellectual Property
Bart Wise, Ph.D., J.D.
Vice President, Intellectual Property
A Multi-Decade Timeline of GalNAc-Mediated Delivery of Nucleic Acids
GalNAc for liver delivery, including plasmids and single- and double-stranded oligos, has a rich, published history
1983
1982
Bi- and triGalNAc
Cluster glycosides
Binding to ASGR
Connolly
et al. 1982
Mono-, bi-, tri-,
tetra-GalNAc
synthetic clusters
and linker
configurations
binding to ASGR
Lee et al. 1983
Carbohydrate
receptors
of the liver
Ashwell &
Harford
1982
1980
GalNAc- & Galpeptide uptake
by hepatocytes
Baenziger &
Fiete 1980
1992
Tri-GalNAc for
ASGR-delivery
to hepatocytes,
YEE(ah-GalNAc)3
synth tripeptide
Lee & Lee 1987
Hexa-GalNAc
& Gal, detailed
linker lengths,
configurations,
binding hepatocytes
Lee RT et al. 1984
1994
Tetra-Gal-poly-Lys
for ASGRmediated delivery
Plank et al. 1992
Oligo-neoglycoprotein
conjugates for
targeted oligo
delivery
Bonfils et al. 1992
ASOR-poly-LLys-oligo
conjugates for
oligo delivery
via ASGR in vitro
Wu & Wu et al. 1992
1988
ASGR-mediated
plasmid gene
delivery in vivo
Wu & Wu
et al. 1988
1980
Tri-GalNAc
gene delivery
to liver
YEE(ah-GalNAc)3
Merwin
et al. 1994
Tri-GalNAc
large scale
production
method
Findeis 1994
Tri-GalNAc >
bi-GalNAc
for liver
targeting
Chiu et al. 1994
1995
Tri-GalNAcoligo
targeted uptake
oligo-conjugate,
YEE(ah-GalNAc)3
Hangeland et al.
(T’so) 1995
Cluster glycoside
synthesis to
bind ASGR on
hepatocytes for
glycotargeting
Biessen et al. 1995
Glycotargeting
via receptors
Wadhwa
& Rice 1995
1981
Cell-specific
ligands selective
drug delivery
Ponpipom
et al. 1981
1990
ASGR model
for endocytic
1987
transport
ASOR-Poly-LSpeiss et al.
Lys-plasmid DNA
1990
delivery
hepatocytes in vitro
Wu & Wu et al. 1987
1997
2001
2003
2002
Multivalent
Tetra-Gal-oligos
Multivalent
Tri-GalNAcGlycoclusters
via branched
GalNac-, Folate32
P-oligo in vivo
in ribozymes
Gal on bases
phosphoramidite
liver uptake
via
dist. along duplex;
Dubber &
YEE(ah-GalNAc)3
phosphoramidites
tri-Gal best
Frechet 2003
Matulic70% after 15 mins
binding ASGR;
Adamic 2002
Hangeland et al.
GalNAc-cluster
via
(T’so) 1997
-oligo ASGR
Cluster
Phosphoramidites
targeting
glycoside
Matsuura
Maier et al.
effect;
et al. 2001
2003
comp review
1999
Lundquist &
Tri-GalNAc-oligo
Toone 2002
conjugates A-L-P
2004
delivery in vivo,
Multivalent
T’so et al. 1999
Glycoclusters:
GalNAc-oligo
Gal on bases
conjugates targeted to
distributed along
parenchymal liver
a duplex
Biessen et al. 1999
binding ASGR;
phosphoramidites
Glycotargeting
nucleic acid; sugar vs
Matsuura
uptake, trafficking,
et al. 2004
Monsigny et al. 1999
1990
Neoglycoprotein
(Gal) uptake
by hepatocytes
Kawaguchi
et al. 1981
55
1984
1987
Glyco-bases, natural,
Gommers et al. 1993
nd
siRNA therapy
review
See Para 4,
Page 3620;
Table 2, Figure 2
on conjugation
Corey 2007
GalNAc/PEG DPC
delivery
of siRNA,
Rozema
et al. 2007
2009
Multi-GalNAcsiRNA
to hepatocytes
in vivo/in vitro
Nair et al. 2014
Non-nucleotide
scaffold GalNAc
Rajeev et al; 2015
2011
GalNAc-oligo
spacer length
affects uptake
Median
et al. 2007
GalNAc-oligos
uptake via
ASGR-mediated
endocytosis
Lönnberg
et al. 2009
2015
Clustered- &
dispersedGalNAc on the
bases & sugars
3’-end siRNA
sense strand
Matsuda
et al. 2015
2013
Multi-valent
sugar moieties,
synthetic or
oligo scaffold
Spinelli
et al. 2013
2010
2000
1993
Tri-Gal-bisacridine
target DNA
to hepatocytes
Haensler & Szoka 1993
2014
Multi-GalNAc2 -gen oligo gapmer
increases
potency in vivo
Prakash
et al. 2014
2007
2014
1997
Tri-GalNAc
synthesis
improvements
[YDD(G-ah-GalNAc)3]
Lee & Lee 1997
1996
Conjugate A-L-P:
A is carbohydrate,
L is a linker and
P is a stable oligo
US 5,994,517
PD 1996, Issue 1999
Expired 2011
T’so et al.
1999
Conjugate A-L-P
GalNAc-oligos
in vivo
review and
new data
Duff et al. 1999
2002
1999
Carbohydrate
conj to linker,
“manifold”,
linker, ligand
US 6,300,319
ISIS Manoharan
PD 1999,
Issue 2001
GalNAcphosphoramidites,
Tri-GalNAc
US 7,491,805
Sirna, Vargeese
PD 2002,
CIP PD 2003,
Issue 2009;
2001
GalNAc-siRNA and
Folate-siRNA,
Increased
bioavailability
US2003/0143732
Fosnaugh
PD 8/2000
2005
Multivalent
glycoclusters
on bases
distributed along
a duplex;
cooperative
ASGR binding
Yamada
et al. 2005
GalNAc-conj
biodegradable
polypeptide
Garrett et al.
2014 (1)
(Merck)
GalNAc-conj
biodegradable
polypeptide
Barrett et al.
2014 (2)
(Merck)
GalNAc-conj
biodegradable
polypeptide
Mouse / NHP
Parmar et al.
2014 (Merck)
Targeting of Oligonucleotides to Liver Via GalNAc Ligands
An early patent filing directed to targeting oligonucleotides, including to liver
• MicroProbe patent filed in 1992
• Directed to oligonucleotides linked through cleavable
peptide linkers to ligands
• Contains disclosure of using a galactose “cluster
ligand” to target oligonucleotides to liver
• “This system is very efficient and well understood.” –
Col. 5, lines 22 - 23.
• Patent was abandoned in 2004
56
Ts’o Patent (priority date = 1995)
What the Examiner said about claims 11 & 17, directed to oligos conjugated to a liver-specific “attachment group”
“Merwin et al teach the well-known ligand for the
asialoglycoprotein receptor, YEE(GalNacAH)3 which ligand is
specific for the liver.” – page 5, Office Action mailed June 24, 1997
[emphasis added]
“It would have been obvious to one of ordinary skill in the art at the
time the invention was made to modify the conjugate of Bonfils et al
with the liver tissues specific ligand of Merwin et al. It would have
been expected, barring evidence to the contrary, that the resulting
conjugate would be effective in delivering substance to the liver.” –
pages 5-6, Office Action mailed June 24, 1997 [emphasis added]
Ts’o et al. eventually obtained claims based on the way they attached
their ligands to the oligonucleotides. However, they abandoned the
patent in 2011.
57
Vargeese Patent (Priority Date = 2002): Example of an Incremental Patent
Issued claims to siRNA conjugated to triantennary carbohydrate ligands using specific linkers
Examiner’s Reasons for Allowance – “Although it is known in
the prior art to conjugate biological molecules such as RNA to
molecules such as N-acetylgalactosamine [GalNAc] cluster
glycosides, . . ., the prior art does not disclose either of the
specific linker compounds 119 or 121 as linkers used to
conjugate two active moieties to one another.” [emphasis
added]
Linker 119
58
Linker 121
Note: Claims 1 and 3 (with Linker 119) were cancelled in a
reexamination at the U.S. Patent Office in 2010 as obvious over
the prior art. One basis for invalidity was the Ts’o patent.
Claims 2 and 4 (with Linker 121) were maintained as valid.
Dicerna’s Patent Portfolio in This Area
• Dicerna has patents and
applications directed to various
Dicerna-developed
oligonucleotide structures, such
as extensions and tetraloops
• Dicerna recently published PCT
application directed to, among
other things, the approaches
Dicerna uses to conjugate
GalNAc ligands to our tetraloop
structures (GalXC platform)
• Dicerna patents and
applications directed to specific
targeting sequences (e.g., MYC,
KRAS, HAO-1, beta-catenin,
Serpin A1)
59
Overview of the Opportunity Space
Pankaj Bhargava, M.D.
Chief Medical Officer
Overview of the Opportunity Space
GalXC enables targeting of disease-driving genes in the liver
1.
2.
3.
4.
5.
6.
61
Dicerna’s Emerging Pipeline of Rare Disease Opportunities
Focus Topic on Primary Hyperoxaluria: Dr. Craig Langman
Dicerna DCR-PHsc Program
Dicerna’s Emerging Pipeline of Larger Disease Opportunities
Focus Topic on Chronic Liver Diseases: Dr. Rebecca Wells
Preclinical CLD Data: HMGB1, β-catenin (CTNNB1), GalXC UDT #8
Dicerna’s Opportunities in Rare Diseases
Rare Diseases
• HAO1 targeted
therapy for primary
hyperoxaluria type 1
Dicerna’s strategy is to retain substantial
rights to key rare disease programs
Rare diseases are an ideal case for GalXC-based therapies
•
•
•
•
•
•
High unmet medical need
A single known genetic cause
Easily identifiable patient population
Easily assayed biomarkers
Centers of excellence for treatment
Supportive advocacy organizations
To date Dicerna has qualified 10 rare disease targets
including both first-in-class and validated target opportunities
62
GalXC UDT #1 for Rare Disease
Indications and clinical characteristics
Related family of rare genetic diseases resulting in life-threatening conditions and multi-organ
complications. Various specific disease types differ in their severity and genetic cause; however,
we believe there may be a target common to all disease types.
Assessments
Technical
Commercial
Opportunity
63
Parameters
Investment Thesis
Validation
Target common to multiple rare diseases in the same family each with differing genetic causes
Genetic deletion of target in mice reduces or eliminates major pathology of disease
Biomarker
Single plasma biomarker associated with the disease family
High Unmet Need
No highly efficacious therapeutic options available
Potentially fatal in the absence of major medical intervention
Patient Numbers
Estimated genetic incidence in excess of 10 per million
Competition
Best-in-class opportunity
Challenging target for small molecules and antibodies
Patient Advocacy
Active patient groups and growing disease awareness
GalXC UDT#1 mRNA Silencing vs. Phenotypic Benefit in a GEMM
One SC dose of GalXC UDT #1 is efficacious
R e m a in in g
UDT#1 mRNA in Disease Model Mice
4
1 0 0
6 0 0
H ig h
B io m a r k e r
M e d
P B S )
5 0
L o w
M e d
3
H ig h
4 5 0
D is e a s e
m R N A
# 1
(re l to
U n d is c lo s e d
disease
L o w
7 5
%
UDT#1 Biomarker in Disease Model Mice
2 5
2
1 0
normal
3 0 0
0
0
dose
1
2
W e e k s
3
0
dose
1
2
3
W e e k s
Therapeutic response in a GEMM mouse model is proportional to mRNA gene silencing
• mRNA silencing of 50% yields an approximately 50% decrease in the disease marker
• 85% mRNA target knockdown drives the disease biomarker down to normal levels
• mRNA and biomarker reductions are approximately linear
64
GalXC UDT #7 for Rare Disease
Indications and clinical characteristics
Metabolic disorder causing progressive decline in liver function despite standard of care.
Long-term outcome includes progressive liver fibrosis and cirrhosis.
Assessments
Technical
Commercial
Opportunity
65
Parameters
Investment Thesis
Validation
Clear predictive causality from pathway has been verified using in-house animal models
Biomarkers
Common plasma disease markers combined with emerging imaging techniques
High Unmet Need
No efficacious therapeutic options available
Symptoms and disease management negatively impacts patient and family quality of life
Patient Numbers
Estimated US/EU prevalence of 3,000 - 5,000 patients
Competition
First-in-class opportunity
Challenging target for small molecules and antibodies
GalXC UDT #7 is Efficacious in a Mouse Model of Liver Disease
GalXC UDT #7 efficacy in vivo
GalXC doses
week 4
week 5
week 6
week 12
week 13
week 18
ALT
AST
ALK
Reduced
toxicity
Molecular
disease marker
reduced
Tolerability
marker
normal
Normalized
Reduced
liver damage
Reduced
liver damage
WT
WT
5
5 0
PBS UDT#7
1000
500
500
WT
WT
0
0
PBS UDT#7
1000
A S T ( U /L )
1 0
1 0 0
PBS UDT#7
PBS UDT#7
200
100
WT
0
0
0
A lk a lin e P h o s p h a t a s e ( U /L )
5
1 5
A L T ( U /L )
10
M a r k e r (m g /d L )
15
300
1500
2 0
M a r k e r C o n c e n tr a tio n
% o f T o t a l B o d y W e ig h t
m R N A
P B S )
( r e l to
T a r g e t # 7
%
66
week 11
Marker #2
0
PBS UDT#7
week 10
Disease Marker 1
20
0
week 9
Liver Weight
mRNA
silencing
5 0
week 8
Take down
UDT #7 mRNA
1 0 0
week 7
T o le r a b ility
Chronic hepatic
damage
Liver Disease
PBS UDT#7
PBS UDT#7
GalXC UDT #7: Treatment in a Mouse Model of Liver Disease
Visualization of liver damage; GalXC UDT #7 conjugate efficacy in vivo
Mouse Model
PBS
Human
20X
20X
20X
GalXC UDT#7
20X
67
Significantly improved liver morphology with GalXC treatment
Overview of the Opportunity Space
GalXC enables targeting of disease-driving genes in the liver
1. Dicerna’s Emerging Pipeline of Rare Disease Opportunities
2. Focus Topic on Primary Hyperoxaluria: Dr. Craig Langman
3. Dicerna DCR-PHsc Program
4. Dicerna’s Emerging Pipeline of Larger Disease Opportunities
5. Focus Topic on Chronic Liver Diseases: Dr. Rebecca Wells
6. Preclinical CLD Data: HMGB1, β-catenin (CTNNB1), GalXC UDT #8
68
Craig Langman, M.D.
• Dr. Craig Langman is the Isaac A. Abt, M.D., professor of Kidney Diseases, and
tenured professor of Pediatrics at the Feinberg School of Medicine,
Northwestern University. He is also the head of Kidney Diseases at the Ann and
Robert H. Lurie Children’s Hospital of Chicago, and director of the two Davita
Children’s Dialysis Centers in Chicago.
• Dr. Langman’s funded research focuses on the basic and clinical expression of
inherited or acquired disorders of calcium, phosphorus, vitamin D, and FGF23
metabolism; mechanisms of cardiovascular diseases in children with obesity or
chronic kidney disease; the proteomics of inherited stone disease; inherited
genetic diseases (cystinosis, oxalosis, kidney stones, atypical HUS,
hypophosphatasia); and the rehabilitation of patients around the world with
chronic kidney disease.
• Dr. Langman has published more than 250 articles, reviews and chapters in his
discipline. He received his medical degree from Drexel University College of
Medicine.
69
Primary Hyperoxaluria:
A New Approach
The Future is Now
Craig B. Langman, M.D.
The Issac A. Abt M.D. Professor of Kidney Disease
Feinberg School of Medicine, Northwestern University
70
Disclosures


71
Craig B. Langman, MD, is a consultant to Dicerna Pharmaceuticals,
Inc.
Craig B. Langman, MD, has documented that he has no other
conflicts to resolve
Outline of Discussion



72
What is Primary Hyperoxaluria Type I?
What do we do now for patients with PH-1?
Why is the Dicerna approach the best idea for patients with Primary
Hyperoxaluria?
Body Homeostasis of Oxalate
liver
PH1
Ox
73
Ox
Types of Hyperoxaluria

10Hyperoxaluria





PH-1 (AGXT)
PH-2 (GR/LDH)
PH-3 (HOGA1)
Unknown type
20Hyperoxaluria

Enteric





Normal
Bariatric Surgery
Bile Acid Diseases
Cystic Fibrosis
Vitamin C excess
Ethylene Glycol
Enteric
Idiopathic
Urinary
Oxalate
(mg/gCr/day)
74
10 mg
30mg 40mg
Primary
50mg
75mg
100mg +
Pathways of the Three Types of Primary
Hyperoxaluria
75
Population Genetics of PH



76
Ultra-orphan disease in all populations
The estimated PH prevalence is 3:1,000,000. However, in
approximately 20% to 50% of cases, patients suffer from either
severe renal insufficiency, or recurrent disease after transplantation,
which occurs before diagnosis.
1,000 patients are in registries!
RKSC PH Registry – April 2016





500 patients enrolled; 347 with PH-1
PH1
Median age at first symptom: 4.9 yrs
Median age at diagnosis: 10.4 yrs
Median follow-up: 5.1 yr, mean 8.5 yrs
Contributing centers: Boston Children’s, Sick Kids Toronto, CHOP,
Lurie Children’s Hospital of Chicago, Cincinnati Children’s, Mayo
Clinic, NYU, Wake Forest, Univ Alabama
683 PH1 patients
OxalEurope, Jul 2015
77
Clinical Burden: Age at Symptom Onset
PH-1 (%)
60
40
20
0
PH-2 (%)
0
5
0
5
10 15 20 25 30 35 40 45 50 60 70 75 80 y
60
40
20
0
PH-3 (%)
60
40
20
0
78
10
15
20
25
30
35
40
Age (years)
45
50
55
60
65
70
75
Primary Hyperoxaluria 1 Pathology
Systemic Oxalosis: Bone
Systemic Oxalosis: Bone & Kidney
The abnormal liver metabolism of PH1 patients produces
excess oxalate concentrated in the renal filtrate
Calcium oxalate crystals form, inducing nephrocalcinosis
Systemic Oxalosis: Skin
Systemic Oxalosis: Kidney
Systemic Oxalosis: Eye
Subsequent decline in kidney function results in systemic
oxalosis with Plasma [Ox] > 30 micromolar
(crystal deposition in other tissues)
Approximately 50% of PH1 patients will have kidney
failure by age 30-35
Photographs reprinted by permission from Macmillan Publishers Ltd:
Nature Reviews Nephrology 8 (2012) pg. 467
79
eGFR at Diagnosis is Already
Compromised in PH-1
CKD Stage
eGFR mL/min/1.73 m2
120
100
80
1
60
2
40
3
20
0
4
PH 1
PH 2
PH 3
Type of PH
80
Clin J Am Soc Nephrol. 2016 Jan 7;11(1):119-26.
Analysis of time to ESRD after PH Diagnosis
100
Free of ESRD (%)
PH3
80
PH2
60
40
PH1
20
P=0.018
n=298 without RF at PH Dx
0
0
10
20
Years after diagnosis
81
30
Unpublished Data
Clin J Am Soc Nephrol. 2016 Jan 7;11(1):119-26.
Nephrocalcinosis
Nephrocalcinosis (%)
70
60
PH 1
50
40
PH 2
30
20
10
PH 3
0
0
5
10
15
20
Years after first image
Incidence (no. at risk)
PH 1
30 (170)
PH 2
16 (32)
PH 3
3.3 (33)
82
40 (43)
32 (8)
7.1 (11)
48 (28)
32 (7)
7.1 (8)
51 (17)
32 (8)
7.1 (5)
57 (11)
32 (8)
7.1 (2)
Kidney Int. 2015 87:623-31. doi: 10.1038/ki.2014.298
Outline of Discussion



83
What is Primary Hyperoxaluria Type I?
What do we do now for patients with PH-1?
Why is the Dicerna approach the best idea for patients with Primary
Hyperoxaluria?
Vitamin B6 Therapy


It has been known for many years that pyridoxine, a metabolic
precursor of the cofactor PLP, can reduce oxalate excretion in about
one third of PH-1 patients.
Pyridoxine responsiveness is associated with two particular
missense mutations in the AGT gene (AGXT) which lead to
Gly170Arg and Phe152Ile amino acid replacements


84
Frequency of 30–40% in PH1 patients in the Western World
Pyridoxine has multiple effects on the Gly170Arg mutant, including
increasing its net expression, catalytic activity, and peroxisomal
targeting. Some of these effects are also found with other mutant
AGTs.
Magnitude of Vitamin B6 Response is
Genotype Dependents in PH-1, but Does Not
Normalize Urinary Oxalate Excretion
best
normal
85
Current Best Rx: Enzyme Replacement Therapy
with a Total Hepatectomy Followed by a Liver
Transplant
86
Urine Oxalate May Remain Elevated Even
After Liver Transplant in PH-1
87
Outline of Discussion



88
What is Primary Hyperoxaluria Type I?
What do we do now for patients with PH-1?
Why is the Dicerna approach the best idea for patients with Primary
Hyperoxaluria?
Substrate Reduction Rx for PH-1
89
Dicerna Treatment Approach
90

Blocking glycolate oxidase (GO)
enzyme lowers the production of
oxalate in the liver → substrate
reduction therapy (SRT); glycolate is
a harmless metabolite and is
excreted in urine

Using RNA interference (RNAi) to
target HAO1 mRNA that encodes GO
is a novel mode of action compared
to existing treatments
Human with HAO1 Deficiency Had No
Urinary Symptoms or Signs




91
Urinary glycolic acid
secretion was 2,000
mmol/mol creatinine (normal,
18-92)
Normal sized kidneys
No stones or
nephrocalcinosis
Normal kidney function
The Rodent Model: the GO
Knockout Mouse

GO -/-
GO-Deficient Mice Develop Normally and Show Glycolic Aciduria
GO -/-
GO -/AGXT -/-
92
Inhibition of Glycolate Oxidase with Dicer-substrate
siRNA Reduces Calcium Oxalate Deposition in a Mouse
Model of Primary Hyperoxaluria Type 1
mRNA
protein
93
Summary and Conclusions





94
PH-1 is a rare, severe, debilitating disease that leads to kidney
failure at a young age.
Treatment is generally unsatisfactory, unpredictable in B6
responsiveness, leading to an unmet medical need.
Liver and/or kidney transplant has its own substantial morbidity and
even mortality associated with it.
RNAi therapy directed at substrate reduction provides a new
treatment approach.
Dicerna is conducting clinical trials now in PH-1 patients.
Overview of the Opportunity Space
GalXC enables targeting of disease-driving genes in the liver
1. Dicerna’s Emerging Pipeline of Rare Disease Opportunities
2. Focus Topic on Primary Hyperoxaluria: Dr. Craig Langman
3. Dicerna DCR-PHsc Program
4. Dicerna’s Emerging Pipeline of Larger Disease Opportunities
5. Focus Topic on Chronic Liver Diseases: Dr. Rebecca Wells
6. Preclinical CLD Data: HMGB1, β-catenin (CTNNB1), GalXC UDT #8
95
GalXC HAO1 is Efficacious in PH1 Mouse Model
GalXC HAO1 provides a subcutaneously injected development option for patients with PH1
A single SC dose of an GalXC HAO1 produced strong effects on urinary biomarkers in
Agxt-/- PH1 disease model mice
Oxalate Reduction
Glycolate Elevation
3000
800
600
400
200
1 m g /k g
2400
2100
1800
1500
1200
900
600
1 m g /k g
300
3 m g /k g
3 m g /k g
W eeks
0
dose
1
2
3
4
W eeks
0
0
96
( m g /g C r e a tin in e /2 4 h r )
2700
U r in e G ly c o la te
O x a la te /C r e a tin in e (m g /g )
1000
5
6
7
0
1
2
3
4
5
6
7
DCR-PHsc (GalXC) Development Plan
• DCR-PHsc IND/CTA filing is expected in late 2017
• The development plan for DCR-PHsc will take into consideration:
-
Safety and biomarker data from the ongoing DCR-PH1 (LNP) clinical trials
Preclinical efficacy and toxicology data
Competitive environment and insights from KOLs and patients
• The ongoing Primary HYperoxaluria Observational Study (PHYOS) will provide important data regarding:
-
97
Natural History and clinical course of patients with PH1
Urine and plasma biomarkers
Quality of life and economic burden of disease
16 patients have been enrolled to date
Overview of the Opportunity Space
GalXC enables targeting of disease-driving genes in the liver
1. Dicerna’s Emerging Pipeline of Rare Disease Opportunities
2. Focus topic on Primary Hyperoxaluria: Dr. Craig Langman
3. Dicerna DCR-PHsc Program
4. Dicerna’s Emerging Pipeline of Larger Disease Opportunities
5. Focus Topic on Chronic Liver Diseases
6. Preclinical CLD Data: HMGB1, β-catenin (CTNNB1), GalXC UDT #8
98
Dicerna’s Opportunities in Large Population Disorders
Chronic Liver
Diseases
• HMGB1 for fibrotic
liver diseases
Cardiovascular
Diseases
• PCSK9 targeted
therapy for hypercholesterolemia
Liver Infectious
Diseases
• HBV targeted
therapy for chronic
hepatitis B infection
Dicerna’s strategy is to work with partners to develop
therapies for large patient populations
GalXC-based therapies can have a major impact
• Simple, infrequent SC dosing paradigm may bring
convenience and compliance benefits
• Exquisite target and tissue specificity nearly
eliminates off-target effects
• GalXC therapy has demonstrated a high
therapeutic index in animal studies
To date Dicerna has qualified 19 disease targets in
these major therapeutic areas
Some targets are relevant to rare disease
subpopulations
99
GalXC PCSK9: Atherosclerotic Cardiovascular Disease (CVD)
Unmet medical need to reduce recurrent CV events in statin-refractory CVD patients
Market Opportunity:
Competition:
35M statin treated patients in U.S.
with hypercholesterolemia1,2
PCSK9 GalXC is being developed for the $10B+ statin-refractory CVD market with the goal of
improved convenience and adherence (less frequent dosing) compared to mAbs.
23M (2/3) without CVD
12M (1/3) with CVD1,2,3
Partnering Opportunities:
Addressable U.S. Patient Population:
> 4.5M (>37%) CVD patients not at LDL
goal of 70 mg/dL on statin alone4
100
1.
2.
3.
4.
5.
Qiuping Gu, et al. NCHS Data Brief. 2014, No. 177.
CDC. Age and Sex Composition in the United States, 2012.
CDC. Summary Health Statistics: National Health Interview Survey 2014.
Wiviott SD, et al. J Am Coll Cardiol. 2005;46(8):1411-1416
Cannon CP, et al. NEJM. 2015; 372 (25).
PCSK9 agents complement portfolios that include other classes shown to reduce CV events;
many pharmaceutical companies would benefit from having PCSK9 GalXC, especially those with
marketed or late-stage mAbs that are likely to be disrupted by emerging long-duration oligos.
6. Eliano PN, et al. Annals of Internal Medicine. 2015; 163 (1).
7. Unpublished data. Alnylam Pharmaceuticals; 11.11.15 Press Release.
GalXC May Play a Key Role in Establishing a Functional HBV Cure
Over 350 million people are chronically infected worldwide
101
Current HBV Therapies Are Insufficient
• Nucleos(t)ide analogs: Even the best NUCs
require lifelong therapy to reduce plasma
viremia, but are ineffective in the long term
since they do not inhibit HBsAg
• PEG-interferon: Poorly tolerated, but a slightly
greater chance of functional cure (~5% of
patients/year)
• Functional Cure of chronic HBV is the best
treatment outcome
- Defined by the lack of detectable HBsAg
(often associated with seroconversion
to anti-HBsAg+)
- A therapy-induced >0.5 log decrease in
HBsAg has positive predictive value for
eventual functional cure (Vigano et al, J.
Gastroenterol 2012)
Organization of the HBV Genome: Overlapping,
Polycistronic mRNA Targets
3,221 1
preS1,
preS2
and S
2,856
Polymerase
EcoRI
2,458
834
2,309
preCore,
Core
1,622
1,816
1,873
X gene
1,376
Overview of the Opportunity Space
GalXC enables targeting of disease-driving genes in the liver
1.
2.
3.
4.
5.
6.
102
Dicerna’s Emerging Pipeline of Rare Disease Opportunities
Focus Topic on Primary Hyperoxaluria: Dr. Craig Langman
Dicerna DCR-PHsc Program
Dicerna’s Emerging Pipeline of Larger Disease Opportunities
Focus Topic on Chronic Liver Diseases: Dr. Rebecca Wells
Preclinical CLD Data: HMGB1, β-catenin (CTNNB1), GalXC UDT #8
Rebecca Wells, M.D.
• Rebecca Wells is an Associate Professor of Medicine, Division of
103
Gastroenterology, Department of Medicine, Perelman School of Medicine,
University of Pennsylvania
• Dr. Wells is a member of the faculty and physician-scientist at the Perelman
School of Medicine at the University of Pennsylvania. She is a member of
the Clinical Practices of the University of Pennsylvania and sees patients at
Penn Medicine at Radnor.
• Her research focuses on the mechanism of hepatic fibrosis, including the
role of liver stiffness, other mechanical factors and extracellular matrix
proteins in fibrosis and cirrhosis; the etiology and mechanism of injury and
fibrosis in biliary atresia; and the role of the matrix and mechanics in
hepatocellular carcinoma.
• Dr. Wells received her bachelor’s degree from Yale University and her
medical degree from Johns Hopkins School of Medicine. She also
completed an internal medicine residency and gastroenterology fellowship
at the Brigham and Women’s Hospital. She has been a gastroenterologist
for 18 years.
Overview:
Chronic Liver Disease and Fibrosis
Rebecca G. Wells
University of Pennsylvania
Disclosures
• Rebecca Wells, M.D., is a consultant to Dicerna Pharmaceuticals,
Inc.
• Rebecca Wells, M.D., has documented that he has no other
conflicts to resolve
Unmet Need in Chronic Liver
Disease and Fibrosis
• 3 million patients in the US with chronic liver disease
• 43,000+ deaths yearly directly from liver disease in the US; 23,000
from liver cancers (together higher than deaths from colon cancer)
• Worldwide, a million deaths/year from cirrhosis (not including liver
cancers)
• Hepatocellular carcinoma, mostly occurring in fibrotic livers, is
second leading cause of death in the world
• $4-5 billion financial burden yearly in the US
Gastroenterology 2015;149:1731–1741
CA: Cancer J Clin 2011;61:69
NIDDK: Digestive Diseases Statistics for the United States
BMC Medicine 2014;12:145
Overall Burden of Liver Fibrosis is
Increasing in the US
• Non-alcoholic fatty liver disease
(NAFLD/NASH) is now the major
cause of chronic liver disease in
the US
• Associated with the metabolic
syndrome (diabetes, obesity)
• Increasing markedly
CGH 2016;14:301
What is Fibrosis?
• Response to injury, generally to
hepatocytes
• Multifactorial etiology, but
common pathways of response
• Abnormal matrix deposition –
scar tissue
• Liver dysfunction results
What is Fibrosis?
• Response to injury, generally to
hepatocytes
• Multifactorial etiology, but
common pathways of response
• Abnormal matrix deposition –
scar tissue
• Liver dysfunction results
Injury to Hepatocytes and
Cholangiocytes Leads to Fibrosis
• Viral
– Hepatitis C
– Hepatitis B
• Fatty
– Non-alcoholic fatty liver disease
– Alcoholic liver disease
• Biliary
– Primary sclerosing cholangitis (PSC)
– Primary biliary cholangitis (PBC)
– Biliary atresia
• Other: autoimmune, inherited metabolic
Many target diseases – pediatric and adult
What is Cirrhosis?
F1
F3
F2
F4
Metavir staging system for liver fibrosis
showing different distribution of matrix
at different stages. Cirrhosis is F4.
What is Cirrhosis?
• End result of liver fibrosis from all etiologies
• Architectural definition: bands of matrix surrounding regenerating
hepatocytes
• Abnormal matrix amount and organization
• Marked hepatocyte and vascular dysfunction
Need to think beyond matrix-depositing cells
when considering therapy
Cirrhosis is Symptomatic
F4
Cirrhosis
Stage 4
Stage 3
Stage 2
Stage 1
F3
F2
F1
F0
Asymptomatic
Fibrosis
• Cirrhosis stages 3-4 symptoms include:
– Variceal bleeding
– Ascites
– Hepatic encephalopathy
• High risk of Hepatocellular Carcinoma
Small changes in fibrosis could have major
clinical benefits
How Are Patients Identified?
• Symptoms of liver failure in patients with end-stage cirrhosis
• Signs and symptoms related to bile duct obstruction in biliary
atresia, PBC
• Population and disease-based screening:
– Baby boomer (1945-1965) hepatitis C screening
– Alcohol abusers
– Obese/diabetic patients
– Patients with ulcerative colitis (PSC)
Can Treatment Impact Fibrosis?
• Extensive evidence that regression occurs (biopsy, stiffness
measurements, biomarkers)
• Even without normalization, significant clinical benefits e.g.
decreased portal HTN
Wanless IR et al, Arch Pathol Lab Med 2000;124:1599
With an effective antifibrotic, patients could
be treated at any stage of disease
Current Therapies
• Targeted elimination of the primary cause of hepatocyte injury
(antiviral therapy, weight loss, cessation of alcohol….)
• No approved direct antifibrotics
• Current clinical trials are focused on single agents
Mechanisms of Fibrosis
• Variable initiating events
• Common cellular elements
– Hepatocytes: source of injury,
drive response
– Hepatic stellate cells: matrix
producing cells
– Immune cells: enhance
response to injury
Hepatocytes as Drivers of Fibrosis
Immune Cells
Injured/Stressed
Hepatocytes
Soluble Factors
Apoptotic Bodies
Hepatocellular
Carcinoma
Hepatic
Stellate
Cells
Dedifferentiated
Hepatocytes
Abnormal
Matrix
Hepatocytes Are Attractive Targets
for Antifibrotic Drugs
• Primary cellular target of injury: prevention
– Viral
– Fatty
– Autoimmune and metabolic
• Soluble factor production: prevent immune response,
prevent and revert stellate cell activation
• Protect against hepatocyte dedifferentiation
• Enhance hepatocyte regeneration
• Direct matrix degrading and remodeling effects
• Appeal of targeting multiple pathways: common and specific
• Potential to reduce the incidence of HCC
Why Target Liver Fibrosis?
• Prevent and slow progression, stimulate regression
symptomatic improvement, decreased complications, decreased
mortality
• Multiple etiologies, affecting multiple populations
• Increasing case and financial burden
Potentially huge impact on health
in US and worldwide
Overview of the Opportunity Space
GalXC enables targeting of disease-driving genes in the liver
1.
2.
3.
4.
5.
6.
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Dicerna’s Emerging Pipeline of Rare Disease Opportunities
Focus Topic on Primary Hyperoxaluria: Dr. Craig Langman
Dicerna DCR-PHsc Program
Dicerna’s Emerging Pipeline of Larger Disease Opportunities
Focus Topic on Chronic Liver Diseases: Dr. Rebecca Wells
Preclinical CLD Data: HMGB1, β-catenin (CTNNB1), GalXC UDT #8
Establishing New Therapies for CLD
GalXC conjugates target hepatocytes exclusively with no pharmacological effect in other cells/tissues
Uncovering New Therapeutic Approaches by
Targeting Hepatocytes
Therapeutic Rationale
• GalXC platform enables exquisite targeting of hepatocytes
•
•
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and the silencing of injury-responsive mRNAs that result in
release of profibrotic damage signals
Dicerna’s platform enables rapid GalXC application to in
vivo disease models:
High fat and other diets mimicking human disease
Mdr2 and other knockouts
Carbon tetrachloride and other hepatotoxicity
models
Complete and partial bile duct ligation
Complex polygenic diseases such as CLD can be effectively
mined to uncover additional hepatocyte targets
NAFLD/NASH, PSC/PBC, AIH, PFIC and other
indications are in scope
Dicerna is investigating multiple additional CLD targets beyond CTNNB1 and HMGB1
Role of HMGB1 in Hepatic Pathology: Mechanism of Action
HMGB1 triggers injury amplification following necrosis
• High-Mobility Group Box 1 (HMGB1) is a highly
abundant and ubiquitously expressed protein that is
conserved from mouse to humans
• HMGB1 neutralizing antibodies:
-
Prevent CCl4-induced acute liver injury1
Protect against hepatic ischemia and
reperfusion injury2
• Conditional deletion of HMGB1 in hepatocytes
decreased APAP-induced neutrophil infiltration and
attenuated liver injury/necrosis and mortality3
Chen et al, 2014
Tsung et al, 2005
3 Huebener et al, 2015
1
2
Chen et al Mol Med 2013
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GalXC HMGB1 Blocks APAP (acetaminophen): Induced Liver Damage
Confirms hepatocyte biology, consistent with published reports using conditional knockout mice
Fast mice
Week
1
2
350 mg/kg
APAP/0.9%NaCl
3
5mg/kg GalXC HMGB1
Dose 1
Dose 2
Dose 3
ALT
Dose 4
Terminate study
24h after APAP
Dose 5
LDH
AST
25000
10000
20000
S a lin e
S a lin e
S a lin e
20000
15000
10000
5000
L D H ( U /L )
A S T ( U /L ) + /- S E M
A L T ( U / L ) + /- S E M
8000
6000
4000
2000
0
124
PBS
G a lX C H M G B 1
10000
5000
0
PBS
15000
0
PBS
PBS
G a lX C H M G B 1
PBS
PBS
G a lX C H M G B 1
GalXC HMGB1 Results in Highly Efficient Silencing in Hepatocytes
Only non-parenchymal cells remain positive for HMGB1 protein
• Potent in vivo mRNA knockdown with murine ED50 ≈1 mg/kg
PBS
10 mg/kg/dose SC, q5dx2, n=5 per cohort
Anti-HMGB1 IHC on FFPE sections, 40X
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GalXC HMBG1
• Kupffer cells, HSCs and LSECs are all expected to express HMGB1
• GalNAc technology can be used to determine if blocking DAMP
signals in hepatocytes alone can prevent HSC activation
β-catenin and CLD
CTNNB1, the gene that encodes β-catenin, is the target for GalXC triggers
β-catenin has multiple direct interactions in liver
beyond the canonical pathway
β-catenin
NFKB
(inflammation)
FOXO
(stress response)
•
•
•
HIF1/2a
(survival/adaptation)
PKA
(liver regeneration)
TCF4
(canonical)
126
• CTNNB1 antisense oligo or GEMM liver knockout is
Integrins/cadherins
(ECM and junctions)
SMAD
(TGFβ-signaling)
Role of Wnt/ β-catenin in CLD is supported by
several recent studies
B-catenin pathway is involved in:
- Zonation
- Regeneration/remodeling
- Homeostasis
- EMT
- Stellate cell activation
•
efficacious in a model of cholestasis.1
CTNNB1 antisense oligonucleotide demonstrated efficacy
in NAFLD model2
LRP6 (Frizzled/Wnt receptor co-activator) loss-of-function
mice are protected from diet-induced steatosis3
β-catenin transcriptional axis directly regulates
transcription of genes involved in lipogenesis,
gluconeogenesis/glycolysis (e.g. G6Pase, PEP
caroboxykinase) 4
β-catenin/Wnt signaling is activated during fibrogenesis in
lung, kidney, liver4
1Nejak-Bowen
et al, AASLD 2014
al, FASEB J 2015
3Go et al, Cell Metabolism 2014
4Monga, Gastroenterology 2015
2Popov et
GalXC β-catenin Prevents Activation of Hepatic Stellate Cells and Liver
Fibrosis in Thioacetamide-Treated Mice
Efficacy achieved with two weekly subcutaneous doses
GalXC
4
3
1
B
+
A
a
A
N
N
iv
lX
C
T
C
A
T
PBS
N
e
Fibrosis readily detectable in liver
5
T
Sirius Red
(Fibrosis)
S
α-SMA
(Stellate cell activation)
necropsy
6
B
TAA
300 mg/kg
7
P
TAA
200 mg/kg
22 25 29
+
11 15 18
A
TAA
100 mg/kg
8
M ean ± S EM
4
Day 29
L iv e r w e ig h t /b o d y w e ig h t ( % )
1
L iv e r : B o d y W e ig h t R a t io
10mg/kg/dose
t= day
G
a
T a rg e t m R N A K n o c k d o w n
60
80% KD
40
20
3500
3000
2500
2000
1500
1000
500
0
C
S
lX
1
B
N
N
T
C
T
A
A
A
+
A
+
G
a
P
a
N
T
G
127
a
lX
C
iv
B
e
C
T
N
P
N
B
B
S
1
0
GalXC
CTNNB1
4000
M ean ± S EM
80
C o l1 a 1 m R N A E x p r e s s io n
L iv e r m R N A E x p r e s s io n
100
M ean ± S EM
C t n n b 1 m R N A E x p r e s s io n
C o lla g e n 1 A 1
120
GalXC β-catenin Protects Mice from a High Fat Diet + TAA Challenge
Demonstration of potential in NAFLD-like setting
G a lX C
β -c a te n in
T h io a c e ta m in e
P ro te c ts
+
H ig h
M ic e
F a t
fro m
L e th a l
D ie t C h a lle n g e
1 1 0
C T N N B 1
P e r c e n t s u r v iv a l
1 0 0
3
9 0
G a lX C
m g /k g /d o s e
(n = 2 0 )
9 5 %
8 0
7 0
U n tre a te d
6 0
(n = 9 0 )
5 0
4 0
5 1 %
3 0
2 0
1 0
0
0
1
2
3
4
5
6
7
8
S tu d y
9
1 0 1 1 1 2 1 3 1 4 1 5 1 6
D a y
High Fat Diet
TAA (100-200 mg/kg)
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GalXC or PBS
(Day -6)
GalXC UDT #8: Multiple Indications
Multiple diseases of different etiologies associated with cholestasis.
Some diseases are rare genetic disorders present from birth, others affect adults.
Severe liver-related disorders are common long-term outcomes despite standard of care.
Assessments
Technical
Commercial
Opportunity
129
Parameters
Investment Thesis
Validation
Clear predictive causality from pathway has been verified using in-house animal models
Biomarkers
Common liver enzyme measurements combined with emerging imaging techniques
High Unmet Need
Inadequate therapeutic options available
Acute symptoms and disease management negatively impacts patient and family quality of
life
Patient Numbers
Estimated combined prevalence in the USA and EU exceeds 100,000 patients
Competition
First-in-class opportunity
Challenging target for small molecules and antibodies
Patient Advocacy
Multiple active groups and growing awareness
GalXC UDT #8: Prevents Liver Damage in Partial Bile Duct Ligation Model
Sirius red staining for collagen after potent silencing of GalXC UDT #8
GalXC
pBDL
GalXC
Week2
Week3
GalXC
Week 4
GalXC UDT #8
PBS
Week1
GalXC
130
4X
4X
4X
4X
4X
4X
4X
4X
4X
4X
4X
4X
• pBDL induced liver fibrosis
• GalXC UDT #8 reduces pBDL induced liver fibrosis
• Variations seen among animals
Selective Lobes Partial Bile Duct Ligation (pBDL)
Quantification of sirius red staining for collagen
% S ir iu s R e d S t a in in g
200
150
100
50
0
Image software
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Animal
A3 1
Animal
A4 2 Animal
B 3 3 Animal
B 4 4 Animal
B5 5
Each data point is represented area selected
from slides sectioned through liver tissue
samples of each animal
Overview of the Opportunity Space in CLD
GalXC enables SC delivery of Dicerna’s RNAi therapies to hepatocytes in the liver
1. Preclinical CLD Data
a) GalXC target hepatocytes exclusively with no pharmacological effect in other cells/tissues
b) GalXC enables exquisite targeting of hepatocytes and the silencing of injury-responsive
mRNAs that result in release of pro-fibrotic damage signals
c) Complex polygenic diseases such as CLD can be effectively mined with Dicerna’s platform to
investigate additional hepatocyte targets and perform rapid testing in preclinical models
2. HMGB1
a) HMGB1 GalXC results in highly efficient silencing in hepatocytes
3. β-catenin (CTNNB1)
a) β-catenin GalXC prevents activation of hepatic stellate cells and liver fibrosis in mice
4. Undisclosed #8
a) Multiple diseases of different etiologies associated with cholestasis
132
Executing Our Strategy
Douglas M. Fambrough, III, Ph.D.
President & Chief Executive Officer
Our Vision
Delivering RNAi-based breakthrough therapies to improve lives
Powerful Capability – durable liver gene silencing with a ≤1ml SC injection
Expansive Opportunity – first-in-class and validated targets across multiple Tx areas
134
Rare Diseases
Chronic Liver
Diseases
Cardiovascular
Diseases
Liver Infectious
Diseases
HAO1
HMGB1
PCSK9
HBV
Primary
hyperoxaluria
type 1
Fibrotic liver
disease
Hypercholesterolemia
Chronic
hepatitis B
infection
Data Presented Today
A selected subset of our accomplishments to date with the GalXC platform
GalXC subcutaneous gene silencing of 12 different disease targets
• 6 examples in non-human primates
• 6 additional examples in rodents
• From idea to potent subcutaneous silencing in rodents in 30 days
GalXC-mediated efficacy for 6 different disease targets
• 3 examples in rare disease models
• 3 examples chronic liver disease models
GalXC platform is now a fully-enabled drug discovery engine
• Potency on par with, or better than, comparable platforms
• Duration of effect supporting monthly or even less frequent dosing
• GalXC infrequent subcutaneous dosing could be superior to daily oral dosing
135
GalXC Development Pipeline – Dicerna Today
Product Candidate
Indication
Stage of Development
2017 Candidates 2016 Candidates
Research
136
DRNA 16.1: DCR-PHsc
Primary Hyperoxaluria
DRNA 16.2: DCR-undisclosed Orphan Genetic Disease
DRNA 16.3: DCR-PCSK9
DRNA 17.1
DRNA 17.2
DRNA 17.3
Cardiovascular Disease
Preclinical
Phase 1
More
Advanced
Studies
Pivotal
Trials
GalXC Development Pipeline Projected Growth – Dicerna 2019
Product Candidate
Indication
Stage of Development
2018 and
2017 Candidates 2016 Candidates
beyond
Research
137
DRNA 16.1: DCR-PHsc
Primary Hyperoxaluria
DRNA 16.2: undisclosed
Orphan Genetic Disease
DRNA 16.3: DCR-PCSK9
Cardiovascular Disease
DRNA 17.1
DRNA 17.2
DRNA 17.3
Dicerna has the capacity to launch 3 new
development programs each year
Preclinical
Phase 1
More
Advanced
Studies
Pivotal
Trials
Executing Our Strategy
Harnessing RNAi’s natural, long-acting, catalytic mechanism for gene silencing
Development Strategy
• Retain substantial rights to high value
programs in focused, monogenic
indications such as rare diseases
• Seek strategic collaborators to develop
therapies for large patient populations
and select rare diseases
Reducing Risk
• Rare genetic diseases where gene silencing
generates a high probability of clinical success
 Primary hyperoxaluria type 1
 Undisclosed genetic metabolic disorders
• Validated targets for disorders with large patient
populations
 PCSK9
 HBV
• Seek strategic collaborations for complex disease
states
 Chronic liver diseases
 Cardiovascular diseases
138
Key Milestones
Company expectations for the next 12-18 months
GalXC Subcutaneous Platform
• DCR-PHsc program launched, IND/CTA late 2017
• Two additional GalXC program launches in 2016 (PCSK9 and undisclosed rare disease)
• Multiple GalXC program launches in 2017
Intravenous Lipid Nanoparticle Clinical Programs
• Primary Hyperoxaluria Type 1
- Patient Phase 1 biomarker readout late 2016/early 2017
- GalXC candidate declared, IND in H2 2017
• Oncology
- DCR-MYC pancreatic neuroendocrine cohort data in 2016
- DCR-MYC biopsy cohort POC data in 2016
- DCR-MYC hepatocellular carcinoma trial data by YE 2016
139