platform advances in rnai therapeutics · 2019-12-14 · 2 agenda welcome • josh brodsky,...

Platform Advances in RNAi Therapeutics

RNAi Roundtable

August 23, 2017

2

Agenda

Welcome

• Josh Brodsky, Associate Director, Investor Relations & Corporate Communications

Introduction

• Akshay Vaishnaw, M.D., Ph.D., Executive Vice President of R&D

Platform advances in RNAi therapeutics

• Advanced ESC Design for Potency Improvements

◦ Vasant Jadhav, Ph.D., Senior Director, Research

• Selection of Well Tolerated GalNAc-siRNAs by Screening in Rodent Toxicity Studies

◦ Maja Janas, Ph.D., Senior Scientist, Early Development

• ESC+ Design with Improved Specificity and Therapeutic Index

◦ Mark Schlegel, Ph.D., Senior Scientist, Research

Q&A Session

3

Reminders

Event will run for approximately 60-75 minutes

Q&A Session at end of presentation

• Submit questions at top of webcast screen

• Questions may be submitted at any time

Replay, slides and transcript available at www.alnylam.com/capella

4

Alnylam Forward Looking Statements

This presentation contains forward-looking statements, within the meaning of Section 27A of the Securities

Act of 1933 and Section 21E of the Securities Exchange Act of 1934. There are a number of important

factors that could cause actual results to differ materially from the results anticipated by these forward

looking statements. These important factors include our ability to discover and develop novel drug

candidates and delivery approaches and successfully demonstrate the efficacy and safety of our product

candidates; pre-clinical and clinical results for our product candidates; actions or advice of regulatory

agencies; delays, interruptions or failures in the manufacture and supply of our product candidates; our

ability to obtain, maintain and protect intellectual property, enforce our intellectual property rights and defend

our patent portfolio; our ability to obtain and maintain regulatory approval, pricing and reimbursement for

products; our progress in establishing a commercial and ex-United States infrastructure; competition from

others using similar technology and developing products for similar uses; our ability to manage our growth

and operating expenses, obtain additional funding to support our business activities and establish and

maintain business alliances; the outcome of litigation; and the risk of government investigations; as well as

those risks more fully discussed in our most recent quarterly report on Form 10-Q under the caption “Risk

Factors.” If one or more of these factors materialize, or if any underlying assumptions prove incorrect, our

actual results, performance or achievements may vary materially from any future results, performance or

achievements expressed or implied by these forward-looking statements. All forward-looking statements

speak only as of the date of this presentation and, except as required by law, we undertake no obligation to

update such statements.

5

Agenda

Welcome


Introduction









Q&A Session

6

HUMAN POC* EARLY STAGE(IND or CTA Filed-Phase 2)

LATE STAGE (Phase 2-Phase 3)

REGISTRATION/

COMMERCIAL

COMMERCIAL

RIGHTS

PatisiranHereditary ATTR

Amyloidosis ● US, Canada,

Western Europe

FitusiranHemophilia and Rare

Bleeding Disorders ●50%

US, Canada,

Western Europe

Inclisiran Hypercholesterolemia ● Milestones &

Royalties

GivosiranAcute Hepatic

Porphyrias ● Global

ALN-CC5Complement-

Mediated Diseases ● Global

ALN-GO1Primary

Hyperoxaluria Type 1 ●Subject to

partner option

rights

ALN-TTRsc02Hereditary ATTR

Amyloidosis ●Subject to

partner option

rights

ALN-HBVHepatitis B Virus

Infection ● Global

Focused in 3 Strategic Therapeutic Areas (STArs):

Genetic Medicines

Cardio-Metabolic Diseases

Hepatic Infectious Diseases

Alnylam Clinical Development Pipeline

*Proof of concept defined as having demonstrated target gene knockdown and/or additional evidence of activity in clinical studies

7

Alnylam Investigational RNAi Therapeutics PlatformExtensive Human Safety Experience

Number of

Programs

Number of

Clinical Studies

Total Patients or

Volunteers Dosed

Greatest Duration

of Exposure

>10 >20 >1000 ~36 months

Platform related findings*

• Low incidence (15.2%) of generally mild, transient injection site reactions

• Low incidence (2.2%) of generally mild, asymptomatic, reversible LFT increases >3x ULN

• No evidence of safety signals similar to revusiran program

Favorable emerging profile for ESC-GalNAc platform compared with

competing oligo platforms†

• No evidence of thrombocytopenia, renal toxicity, or systemic inflammatory effects

*All reported data as of December 2016† Based on reported study data - not based on direct comparative studies

8

GalNAc-Conjugate Platforms for Delivery to Liver

• Standard Template Chemistry

GalNAc conjugate

• SC administration

• Fitusiran

• Inclisiran

• Givosiran

STC-Conjugate ESC-Conjugate

• ALN-TTRSC02• ALN-GO1• ALN-CC5• ALN-HBV

Revusiran

• Enhanced Stability Chemistry

GalNAc conjugate


• 2018 INDs and CTAs

ESC+ Conjugate

• Enhanced Stability Chemistry

GalNAc conjugate, ↑ specificity


First generation

GalNAc conjugate,

Initial human POC

Second generation

GalNAc conjugate,

Human POC,

greater potency

and durability with

lower exposures

Next generation

GalNAc conjugate

with further

improvements to

specificity and

therapeutic index

9

Agenda

Welcome


Introduction









Q&A Session

10

Enhanced Stabilization Chemistry (ESC) GalNAc-siRNAsSC-Administered Conjugate Platform for Targeted Delivery to Hepatocytes

AsialoglycoproteinReceptor (ASGPR)• Highly expressed in

hepatocytes

• High capacity receptor

• Conserved across species

siRNA• Metabolic stability

• Intrinsic potency

Ligand• Trivalent GalNAc

• High affinity and specificity

11

Recap of STC to ESC Transition

Metabolic profiling of conjugates in mouse liver

Standard Template Chemistry (STC)

Enhanced Stabilization Chemistry (ESC)

5′-sense

5′-AS

5′-sense

5′-AS

= Phosphorothioate linkage (PS) = 2’-F = 2’-OMe

Region of nuclease

hotspots

Region of nuclease

hotspots

End stabilization with PS linkages

12

ESC Significantly Enhances Efficacy and DurationReduction of AT Protein After Single SC Dose in NHP

Potent and durable silencing achieved after single SC dose• >10-fold improvement in efficacy over standard template chemistry

• Substantially extended duration of effect

% K

no

ck

do

wn

se

rum

AT

(R

ela

tive

to

pre

-do

se

)

Day

100

80

60

40

20

1.0

1.2

-10 0 10 20 30 40

STC-AT3 (10 mg/kg)

ESC-AT3 (10 mg/kg)

ESC-AT3 (1 mg/kg)

Single SC dose

13

Objective• Identify new generalizable conjugate designs with improved in vivo potency through optimal

placement of 2’-F and 2’-OMe modifications

Approach• Begin with motifs identified by statistical analysis of large dataset of conjugate in vitro activity

• Apply screening platform using a set of multiple standard sequences (3 targets) and conditions for rapid and systematic evaluation of new designs

siRNA design

In vitro activityIn vivo efficacy /

exposure

Generalizability

(expanded screen)

Towards Advanced ESC DesignsIterative Screening Platform to Drive Continuous Design Improvements

14

Performance of Advanced ESC Conjugate in Mice

Advanced ESC Design

0.0001

0.001

0.01

0.1

1

10

4h 3 7 14 21 35

Liv

er

Exp

osu

re (

ug

/g)

Days Post-Dose

ESC Design

2’-F 2’-OMe PS

0.0

0.2

0.4

0.6

0.8

1.0

1.2

4h 3 7 14 21 35

mR

NA

(n

orm

to

PB

S)

Advanced ESC (0.75 mg/kg)

ESC (2.5 mg/kg)

Target mRNA levels in Liver Total siRNA levels in Liver

Days Post-Dose

Advanced ESC (0.75 mg/kg)

ESC (2.5 mg/kg)

15

Performance of Advanced ESC Conjugate in NHP

ALN-TTRsc02 (Advanced ESC design) compared to Revusiran

Revusiran

ALN-TTRsc02

Days

Total

Dose

AUEC for %

TTR lowering

Revusiran 45 mg/kg 4154

ALN-TTRsc02 1 mg/kg 8139

Adjusting for dose difference (45-fold) and AUEC

(1.95-fold), ALN-TTRsc02 shows ~88-fold in vivo

potency improvement over Revusiran

AUEC= Area under effect curve

16

Me

an

[+

/- S

EM

] T

TR

Re

lative

to

Ba

se

line

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

Days since first dose

0 40 80 120 160 200 240 280 320

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

Cohort Placebo (N=20) TTRSC02 (5mg) (N=6)TTRSC02 (25mg) (N=6) TTRSC02 (Optional; 25mg) (N=6)TTRSC02 (Subjects of Japanese descent; 25mg) (N=6) TTRSC02 (50mg) (N=6)TTRSC02 (Optional; 50mg) (N=6) TTRSC02 (Subjects of Japanese descent; 50mg) (N=6)TTRSC02 (100mg) (N=6) TTRSC02 (200mg) (N=6)TTRSC02 (300mg) (N=6)

ALN-TTRsc02 Phase 1 Preliminary Study Results

Single Ascending Dose Study in Healthy Volunteers

• No SAEs and no discontinuations due to AEs

• All AEs mild or moderate in severity

◦ 14 AEs in 8 subjects considered possibly related to treatment; majority mild

◦ Events included injection site erythema, injection site pain, injection site bruising, rhinorrhea, pruritus, cough, nausea, fatigue, genital rash and abdominal pain

◦ No clinically significant changes in lab parameters, EKG or physical exam

As of data cutoff on 31May2017

17

Pushing the EnvelopeSteadfast Focus on Platform Research Driving Potency Improvements

Sharp’s Law

Conjugate potency improvements in mice (single dose ED50)

0.01

0.1

1

10

100

2005 2007 2009 2011 2013 2015 2017

ED

50 (

mg

/kg

)

Year

Endo-

light

STC

ESC

Advanced ESC

Projected

ESC

Advanced

ESC

Endo-light

STC

18

Agenda

Welcome


Introduction









Q&A Session

19

GalNAc-siRNAStudy

Study Population

Subjects Treated

Max Duration Treatment

ALT>3x ULN

Revusiran Phase 1 NHV 66 6 wks 1Revusiran Phase 2 ATTR cardiomyopathy 26 6 wks 1Revusiran Phase 2 OLE ATTR cardiomyopathy 25^ Up to 18 mos 0Fitusiran Phase 1, Part A NHV 3 Single dose 0

Fitusiran Phase 1, Parts B & CSevere and moderate

HA and HB w/o inhibitor 25 Up to 3 mos 1Fitusiran Phase 1 D HA and HB w/ inhibitor 16 Up to 3 mos 3Fitusiran Phase 2 (OLE) HA and HB w/ and w/o inhibitor 23^ Up to 14 mos 3ALN-CC5 Phase 1, Parts A & B NHV 33 Up to 3 wks 0ALN-CC5 Phase 1, Part C PNH 6 Up to 6 mos 1Inclisiran Phase 1 NHV w/ elevated LDL-C 51 Up to 2 mos 1Inclisiran Phase 2 High Risk CVD; elevated LDL 370 Single dose 1Givosiran Phase 1, Parts A & B ASHE 23 Up to 2 mos 1Givosiran Phase 1, Part C AIP 11 Up to 6 mos 1ALN-AAT Phase 1/2, Parts A & B NHV 19 Up to 4 mos 1ALN-GO1 Part A NHV 24 Single dose 0

ALT=alanine aminotransferase; ULN=upper limit of normal ^Subjects treated with drug in previous study

ALT Elevations in Clinical ProgramsData Transfer as of November 2016

LNP-siRNAStudy

Study Population

Subjects Treated


ALT>3x ULN

Patisiran Phase 1 NHV 22 Single dose 0Patisiran Phase 2 hATTR polyneuropathy 29 Up to 4 wks 0Patisiran Phase 2 OLE hATTR polyneuropathy 27^ Up to 25mos 1

STC

ESC

20

GalNAc-siRNAStudy

Study Population

Subjects Treated


ALT>3x ULN

Revusiran Phase 1 NHV 66 6 wks 1Revusiran Phase 2 ATTR cardiomyopathy 26 6 wks 1Revusiran Phase 2 OLE ATTR cardiomyopathy 25^ Up to 18 mos 0Fitusiran Phase 1, Part A NHV 3 Single dose 0

Fitusiran Phase 1, Parts B & CSevere and moderate

HA and HB w/o inhibitor 25 Up to 3 mos 1Fitusiran Phase 1 D HA and HB w/ inhibitor 16 Up to 3 mos 3Fitusiran Phase 2 (OLE) HA and HB w/ and w/o inhibitor 23^ Up to 14 mos 3ALN-CC5 Phase 1, Parts A & B NHV 33 Up to 3 wks 0ALN-CC5 Phase 1, Part C PNH 6 Up to 6 mos 1Inclisiran Phase 1 NHV w/ elevated LDL-C 51 Up to 2 mos 1Inclisiran Phase 2 High Risk CVD; elevated LDL 370 Single dose 1Givosiran Phase 1, Parts A & B ASHE 23 Up to 2 mos 1Givosiran Phase 1, Part C AIP 11 Up to 6 mos 1ALN-AAT Phase 1/2, Parts A & B NHV 19 Up to 4 mos 1ALN-GO1 Part A NHV 24 Single dose 0

ALT Elevations in Clinical ProgramsData Transfer as of November 2016

LNP-siRNAStudy

Study Population

Subjects Treated


ALT>3x ULN

Patisiran Phase 1 NHV 22 Single dose 0Patisiran Phase 2 hATTR polyneuropathy 29 Up to 4 wks 0Patisiran Phase 2 OLE hATTR polyneuropathy 27^ Up to 25mos 1

STC

ESC

Frequency of ALT >3x ULN = 2.2%

(16/724)

ALT=alanine aminotransferase; ULN=upper limit of normal ^Subjects treated with drug in previous study

21

GalNAc-siRNA ConjugatesWide Therapeutic Index from Non-Clinical GLP Studies

NOAEL1 4 or 7 weeks

(mg/kg)

NOAEL 13 weeks

(mg/kg)

NOAEL 6 mos Rat

(mg/kg)

NOAEL 9 mos

NHP (mg/kg)

Expected Human

Therapeutic Index (TI)2

Rat NHP Rat NHP

Revusiran3 30 ≥300 N/A N/A 30 ≥200 >80

ALN-TTRsc02 N/A N/A ≥120 ≥300 Planned Planned >500

Fitusiran3,4 15 0.35 N/A N/A 0.55

0.55

>105

Inclisiran ≥250 ≥250 ≥250 ≥250 ≥250 ≥300 >500

ALN-CC5 ≥50 ≥100 50 100 ≥50 ≥100 >200

Givosiran N/A N/A ≥30 ≥150 Ongoing Ongoing >500

ALN-AAT N/A N/A ≥50 ≥150 Ongoing Ongoing >500

ALN-HBV N/A N/A ≥60 ≥200 Planned Planned>500

1No Adverse Event Level (NOAEL)2TI calculated as NOAEL in NHP/Expected dose in man37 week studies4NOAEL in hemophilia mice >100 mg/kg, qW x 75 Secondary to exaggerated pharmacology

NHP

◦ Liver

– Basophilic granules in Kupffer cells

and hepatocytes; represents drug

accumulation

◦ Lymph nodes

– Vacuolated macrophages (with

basophilic stippling); represents

phagocytosis of drug

Rat

◦ Liver– Hepatocellular vacuolation: increased

number and size of normal rat hepatocellular vacuoles, most contain lipid

– Increased single cell necrosis

– Increased mitosis and regeneration

◦ Kidney– Basophilic granules in proximal tubular

epithelium; represents drug accumulation

22

In silico prediction &In vitro efficacy

In vitro screen for predicted off-targets

Rodent Knockdown

Rat Tox @ >100x PD dose

NHP Knockdown DC

Bad Actors (40%):

Show hepatotoxicity

Good Actors (60%):

No hepatotoxicity

Subset of Conjugates Shows Rat Hepatotoxicity at Toxicological

Doses and Drop Out of DC Selection Process

Single cell necrosis and/or

hepatocellular degeneration with

↑LFT 2x upper limit of normal

23

Potential Causes of Hepatotoxicity in Rodent Toxicity Studies

1. Non-RNAi drug effects e.g. protein binding

2. Competition for RISC

loading with miRNAs

RISC loading

mRNA

RISC

On-target bindingFull sequence match

Desired

on-target activity

3’-UTR

Off-target bindingPartial sequence match

3. Undesired off-target

activity

24

5’ Modifications on Antisense Strand Can Block

RISC Loading With No Impact on Liver Exposure

RISC loading

mRNA

Sense strand removal

Target RNA cleavage

Dose: 30 mg/kg

Regimen: q2d x 6

Necropsy: Day 15

N o Y e s N o Y e s

1

1 0

1 0 0

R a t L iv e r E x p o s u re

ug

AS

/ g

liv

er

s iR N A -1 s iR N A -2

R IS C lo a d in g b lo ckN o Y e s N o Y e s

0 .0

0 .2

0 .4

0 .6

R a t R IS C L o a d in g

ng

AS

/ g

liv

er


R IS C lo a d in g b lo ck

25

Blocking Antisense RISC Loading Without Altering

2’F/2’OMe/PS Content Mitigates Hepatotoxicity

*

*^

#

No Yes

RISC loading block (siRNA-1)

Dose: 30 mg/kg

Regimen: q2d x 6

Necropsy: Day 15

N o Y e s N o Y e s

0

1 0 0

2 0 0

3 0 0

R a t A L T

U/L

R e fe re n c e ra n g e


R IS C lo a d in g b lo ck

26

Accumulation of chemically-modified siRNAs in the liver is not

sufficient for rat hepatotoxicity.

27

ESC

Advanced ESC

Changing siRNA Chemical Modification Pattern

Does Not Reduce Liver Exposure or RISC Loading Dose: 100 mg/kg

Regimen: qw x 9

Necropsy: Day 58

E S C Ad v a n c e d E S C

0

1 0 0

2 0 0

3 0 0

4 0 0

5 0 0


ng

AS

/ g

liv

er

E S C Ad v a n c e d E S C

1

1 0

1 0 0

1 0 0 0


ug

AS

/ g

liv

er

28

Changing siRNA Chemical Modification Pattern

Does Not Mitigate Hepatotoxicity Dose: 100 mg/kg

Regimen: qw x 9

Necropsy: Day 58

S a lin e E S C Ad v a n c e d E S C

0

1 0 0

2 0 0

3 0 0

R a t A L T

U/L


ESC Advanced ESC

#

* #*

29

siRNA sequence, not the chemical modifications, is important for rat

hepatotoxicity.

30


1. Non-RNAi drug effects e.g. protein binding


loading with miRNAs

RISC loading

mRNA

RISC


Desired

on-target activity

3’-UTR



activity

31

Reversir to Abrogate RNAi Activity Without Changing siRNA

Chemistry or RISC Loading

0%

20%

40%

60%

80%

100%

120%

0 3 6 9 12 15 18 21 24

Rel

ativ

e Ta

rget

Pro

tein

Lev

el

No Reversir

Time (Days)

3 mg/kg

GalNAc-siRNA

0.1 mg/kg

GalNAc-Reversir

Reversir

Reversir

Reversir binds as a “synthetic target” to antisense strand in functional RISC:

32

Prevention

Treatment

Reversir

siRNA

Reversir

siRNA

( )

Blocking RISC-Loaded Antisense Strand with Reversir

Does Not Reduce Liver Exposure or RISC Loading

3-10 mg/kg

30 mg/kg

3-10 mg/kg

30 mg/kg

1

1 0

1 0 0

1 0 0 0


ug

A

S /

g l

ive

r

- Targeting Ctr - Targeting Ctr Targeting Ctr RVR

siRNA1 2 3

1

1 0

1 0 0

1 0 0 0


ng

A

S /

g l

ive

r

- Targeting Ctr - Targeting Ctr Targeting Ctr RVR

siRNA1 2 3

33

Reversing Activity of RISC-Loaded Antisense Strand

Mitigates Hepatotoxicity

Pre

ve

nti

on

+ Ctr RVR + Targeting RVR

#*

#

+

*

*#

**

*

^

Tre

atm

en

t

0

5 0

1 0 0

1 5 0

R a t G L D H

U/L


- Targeting - Targeting Ctr RVR

siRNA- 1

0

1 0

2 0

3 0

4 0

R a t G L D H

U/L



siRNA- 2

0

1 0

2 0

3 0

4 0

R a t G L D H

U/L



siRNA- 3

siRNA: 30 mg/kg qw x 3

Reversir: 3 or 10 mg/kg SD

Necropsy: Day 15 (Prevention)

Day 30 (Treatment)

34

Competition for RISC loading with endogenous miRNAs is not a major

contributor to rat hepatotoxicity; seed-mediated hybridization of the

RISC-loaded antisense strand appears to be important.

35



loading with miRNAs

RISC loading

mRNA

RISC


Desired

on-target activity

3’-UTR



activity

36

Seed-Mediated Off-Target Effects Observed at High Doses

RNAseq in Rat Hepatocytes (24 hrs, 10 nM)

siRNA-2siRNA-1

Downregulated genes Upregulated genes

Antisense seed

enrichment

Sense seed

enrichment

Antisense seed

enrichment

Sense seed

enrichment

siRNA-1 p < 2.2e-16 p = 0.05286 p = 0.667 0.2174

siRNA-2 p < 2.2e-16 p = 0.9679 p = 0.6345 p = 0.3364

65

199

160

190

p < 0.05

On-target mRNA

On-target mRNA

37

ESC GalNAc-siRNA Hepatotoxicity Summary

Antisense strand-driven RNAi-mediated hybridization-based off-target

effects, not chemical modifications or competition for RISC loading with

miRNAs, appear to be important drivers of rat hepatotoxicity.

3’-UTR


Translation repression,

mRNA destabilization

3’-UTR

Undesired off-target activity

38

Agenda

Welcome


Introduction









Q&A Session

39

ESC+ Strategy for Improved Specificity and Therapeutic Index

Utilize seed-pairing destabilization via novel chemical

modifications to selectively destabilize off-target binding

Off-target mRNA 3’-UTR


Antisense loaded RISCpos. 2-8

Undesired off-

target activity

40

ESC+ Conjugates

Design Considerations

Important Considerations

1. On-target potency must be maintained in vivo

2. Minimal impact on metabolic stability

3. Off-target activity should be minimized

Position and nature of modification important for all three

Incorporate modification(s) to

destabilize seed-pairing

41

ESC+ vs. ESCComparable Potency and Reduced Off-Target Activity In Vitro

AD-64958 IC50: 0.0115 nM

Concentration in [nM]1x10

-50.001 0.1 10

%T

arg

et

rem

ain

ing

0

20

40

60

80

100

120

AD-72788 IC50: 0.0041 nM

Concentration in [nM]1x10

-50.001 0.1 10

%T

arg

et

rem

ain

ing

0

20

40

60

80

100

120

ESC

ESC+

Potency Off-Target Activity

On-target

On-target

42

ESC+ vs. ESC

Comparable Potency in Rats

ED50 ~ 0.050 mg/kg ED50 ~ 0.075 mg/kg

ESC ESC+

0

0.2

0.4

0.6

0.8

1

1.2

1.4

-4 0 4 8 12 16 20 24 28 32 36 40 44 48 52 56 60

Seru

m T

TR r

elat

ive

to D

ay 0

Day post-dose

0.075 mg/kg

0.15 mg/kg

0.3 mg/kg

1 mg/kg

0

0.2

0.4

0.6

0.8

1

1.2

1.4

-4 0 4 8 12 16 20 24 28 32 36 40 44 48 52 56 60

Seru

m T

TR r

elat

ive

to D

ay 0

Day post-dose

0.075 mg/kg

0.15 mg/kg

0.3 mg/kg

1 mg/kg

43

ESC+ vs. ESC

Hepatotoxicity Mitigated in Rats

Degeneration 4/4 (2.5)

SCN 4/4, (1.25)

Coag Nec 1/4 (1)

Increased mitoses 3/4 (2.25)

Bile duct hyperplasia 1/4 (1)

Vacuolation 4/4 (2)

Degeneration 0/4

SCN 1/4, (1)

Coag Nec 0/4

Increased mitoses 0/4

Bile duct hyperplasia 0/4

Vacuolation 3/4 (1)

ESC ESC+

Dose: 30 mg/kg

Regimen: qw x 3

Necropsy: Day 15

44

ESC+ vs. ESC

≥6-Fold Improvement in Therapeutic Index in Rats

ESC

0.0

75

0.1

5

0.3

0.6 1 3

10

30

60

12

0

0

2 5

5 0

7 5

1 0 0

0

1

2

3

4

5

A D -6 4 9 5 8

D o s e (m g /k g )

% K

no

ck

do

wn

To

x g

ra

de

N O A E L

N O E L

E D8 0 T I =

66

ESC+

0.0

75

0.1

5

0.3

0.6 1 3

10

30

60

12

0

0

2 5

5 0

7 5

1 0 0

0

1

2

3

4

5

A D -7 2 7 8 8

D o s e (m g /k g )

% K

no

ck

do

wn

To

x g

ra

de

N O A E L

N O E L

E D8 0 T I 4 0 0

45

Placebo

0.1 mg/kg

0.3 mg/kg

1.0 mg/kg

3.0 mg/kg

6.0 mg/kg

ALT AST

-10 0 10 20 30 40 50 60 70

Days Since Dose

-10 0 10 20 30 40 50 60 70

Days Since Dose

1X ULN

ALN-AAT Phase I Single Dose in Healthy VolunteersSerum Transaminases Elevations at 3 and 6 mg/kg SD

N

5

3

3

3

3

3

1

4

8

1

4

8

1

4

8

1

4

8

1

4

8

1

4

8

Data transfer: 30Jun2016

Haslett et al., OTS, September 2016

46

ALN-AAT02: ESC+ Version of ALN-AAT with Reduced

Off-Target Activity In Vitro

ALN-AAT ALN-AAT02

-94%

0

52AAT mRNA

0

3AAT mRNA

RNAseq from Hep3B cells transfected with 10 nM siRNA, 16 hrs treatment

47

ALN-AAT02 Retains Potent Activity in NHP (1 mg/kg)

Plan to advance ESC+ ALN-AAT02 into clinical development in 2018

0 2 0 4 0 6 0 8 0 1 0 0

0 .0

0 .2

0 .4

0 .6

0 .8

1 .0

1 .2

1 .4

D a y s P o s t-D o s e

AA

T (

Re

lati

ve

to

Pre

-do

se

)A L N -A A T

A L N -A A T 0 2

48

Summary

• Platform advances continue to drive potency of GalNAc-siRNA conjugates,

potentially allowing quarterly or even bi-annual dosing at low doses in

humans

• Demonstrated RNAi-mediated off-target effects as important driver of

hepatotoxicity for subset of ESC conjugates in rodent toxicity studies

◦ No evidence for impact of chemical modifications on observed toxicity

• Developed ESC+ design for mitigating seed-mediated off-target effects

◦ Improves specificity and further expands therapeutic index of conjugates

• Plan to advance first ESC+ conjugate, ALN-AAT02, into clinical

development in 2018

• All future candidates planned to employ ESC+ design

49

Agenda

Welcome


Introduction









Q&A Session

50

Upcoming RNAi Roundtables

Givosiran, in development for the treatment of acute hepatic porphyrias

• Thursday, September 7, 10:30 am ET

Fitusiran, in development for the treatment of hemophilia and rare

bleeding disorders

• Tuesday, September 12, 10:30 am ET

Additional details for upcoming RNAi Roundtables, including speakers, dates and times, will be

provided on the Capella section of the Company's website, www.alnylam.com/capella.

http://www.alnylam.com/our-science/capella/

Thank you

platform advances in rnai therapeutics · 2019-12-14 · 2 agenda welcome • josh brodsky,...

Documents