Improved outcomes after intratumoral administration of immunostimulatory mRNA in a novel cationic lipid

Slide 1 Confidential and Proprietary · © 2016 OnkaidoConfidential and Proprietary · © 2016 Onkaido

Improved outcomes after intratumoral administration of immunostimulatory mRNA in a novel cationic lipid4th International mRNA Health ConferenceNovember 1, 2016; Boston, MA Steve Kelsey MD, President, Onkaido, a Moderna venture

Slide 2 Confidential and Proprietary · © 2016 Onkaido

onkaido------------a moderna venture

ONCOLOGY

About Onkaido• Formed 2014• Wholly-owned Moderna Venture• mRNA cancer therapeutics• Fully integrated leverage of

Moderna platform and ecosystem


Cancer Immunotherapy Coming of Age

Coley’s toxins BCG IL2Checkpoint inhibitors IFNaImmunosurveillance

theoryAllogeneic

BMT

1891 1957 1970s 1986 1995 1998 2010-2015

Cancer vaccines


Potential for intratumoral injection of mRNA encoding immunostimulatory molecules

• Combining immunostimulatory proteins with checkpoint inhibitors may improve outcomes from cancer therapy

• Direct injection of mRNA encoding immunostimulatory molecules such as OX40L has huge potential in the treatment of amenable tumors

• Immunosuppressive effects of cytokines induced by lipid nanoparticles, or pre-medication, are counterproductive in the context of immunotherapy

Yao et al Nature Rev Drug Discov 2013


Why messenger RNA for cancer?

Potential advantages over conventional

therapeutics

Potential advantages over gene therapy

• ‘Undruggable’ targets with intra-cellular proteins• Induction of unwanted peptides or proteins• Induction of transmembrane proteins• Repletion of missing proteins• Vaccines

• Transient, regulatable effect• Does not need to enter the nucleus• No risk of genomic integration

• Easily enabled combinations• Rapid research turnaround time


• Lipid nanoparticles remain the most tractable method of delivering mRNA in vivo

• Cationic lipids such as Dlin-DMA and derivatives (eg. Dlin-MC3-DMA) are associated with toxicity in vivo

• Complement activation and cytokine induction, such as IL6, is observed

• Premedication with corticosteroids is frequently used in the clinic

From: Tabernero et al, Cancer Discovery, 2013


Moderna has invested heavily in discovery and development of novel LNP components

• 2+ year effort• >30 FTEs• chemistry, formulation, drug product development, pharmacology, bioanalytics

Relative protein expression in tumors after intratumoral injection of LNP-formulated mRNA

mRNA formulated at Onkaido TherapeuticsStudies performed at Medicilon Inc, Shanghai

GFP expression in A20 Tumor2.5 ug/mouse, iTu, 6 hours

PBS

MC3

SM10

0SM

102

SM10

3SM

104

SM10

7SM

113

SM12

2SM

123

SM12

8SM

140

SM15

1SM

157

SM18

0SM

187

SM22

3SM

233

SM23

4M

C3SM

28SM

144

SM17

5SM

191

SM19

2SM

193

SM21

2SM

224

SM24

6SM

247

SM24

8SM

249

SM25

2SM

253

SM25

4M

C3SM

86SM

217

SM22

1SM

225

SM22

6SM

227

SM22

8SM

251

SM25

5SM

259

SM26

2SM

263

SM26

4SM

265

SM26

6SM

267

SM26

8SM

269

MC3

SM86

SM02

7SM

034

SM07

3SM

085

SM09

7SM

143

SM17

4SM

178

SM27

1SM

287

SM28

8SM

289

SM29

0SM

012

SM21

2SM

175

SM22

0M

C3 (D

PBS)

SM86

(tris

)0

2

4

6

8

10

GFP

/tiss

ue(N

orm

aliz

ed to

MC

3)GF

P pe

r gra

m tu

mor

no

rmal

ized

to st

anda

rd

Stan

dard

Stan

dard

Stan

dard

Stan

dard

N1G

L


N1GL, a novel cationic lipid, has a favorable profile on initial in vivo screening

8.0

7.0

6.0

5.0

4.0

3.0

2.0

1.0

0

100

50

0

Protein expression

Tumor IL6 levels

Protein to IL6 ratio

N04GL N51GL N22GL N1GL N04GL N51GL N22GL N1GL

Studies performed at Medicilon Inc, Shanghai

Rela

tive

tum

or le

vels


N1GL has a short plasma and tissue half life

100

1000

1E+04

1E+05

0 5 10 15

Mean vs TimeMC3SM-86

0 4 8 12 16 20 240

100

200

300

400

500

SM-86 liver

Time (h)

ng/g

tiss

ue

Day 1

Day 8

Day 15

Plasma lipid after a single IV infusion NHP

N1GL in liver 24hrs after weekly dosing

Tissue PK in mice

Studies performed at Charles River Labs, and Agilux Labs

N1GL in plasmaAfter single IV dose


N1GL-based LNPs are well tolerated after local injection to rodents and primates

Toxicology findings

Injection site reactions

Systemic inflammation

Systemic inflammation induced stress

Secondary findings

STD10 (mg RNA/kg) 1

HNSTD (mg RNA/kg) 0.3

Studies performed at Charles River Laboratories


Intratumoral injection of mRNA in N1GL results in 20-25% of transfected cells, including tumor and myeloid populations

A20 tumor modelAnalyzed by flow cytometry 24hrs after dosingStudies and analysis performed at Onkaido Therapeutics

Control 0.5ug 2.5ug 12.5ug0

5

10

15

20

25

Proportion of Transmembrane Targetexpressed across cell type

% o

f liv

e ce

lls

CD4+ T Cells

CD8+ T Cells

CD11b+ Myeloid CellsNK Cells

A20 Cancer CellsB Cells

Non-cancer/non-immune cells

N1GLMajority of expression in tumor cells and myeloid cells


Minimal G-CSF induction with N1GL

NT 0.5ug 2.5ug0.0

0.5

1.0

1.5

2.0

Tumor G-CSF

Con

cent

ratio

n (n

g/g

tum

or)

6hr24hr

N1GLNT 0.5ug 2.5ug

0

10

20

30

40

Plasma G-CSF

Con

cent

ratio

n (p

g/m

L)

6hr24hr

N1GL

A20 tumor modelStudies and analysis performed at Onkaido Therapeutics


mRNA in N1GL associated with minimal increase in tumor myeloid-derived suppressor cells (MDSCs)

PBS 0.5ug 2.5ug 12.5ug0.0

0.5

1.0

1.5

Average Ly6G+ cells (PMN-MDSCs / neutrophils)%

of l

ive

cells

N1GLA20 tumor modelAnalyzed by flow cytometry 24hrs after dosingStudies and analysis performed at Onkaido Therapeutics


Single agent OX40L formulated in N1GL is efficacious

10 20 30 40 50 60 70 80 90 100 1100

500

1000

1500

2000

SM86 2.5g Transmembrane Target

Days post implant

2 CRs

Individual tumor volumes in MC38 tumor model with weekly intratumoral injection of mRNA encoding murine OX40LStudy performed at Onkaido Therapeutics

N1GL 2.5ug OX40L mRNA


Overcoming resistance to anti-PD1 therapy THREE PATIENT POPULATIONS:

• Primary failures: 60% progress within 6 months

• Partial responders: 23%, many progress

• Complete responders: 10%

3yr follow-up for Keytruda in melanoma Robert et al, ASCO 2016

Anitei et al, Clin Cancer Res 2014


Leveraging multiple entry points and combinations to enhance the cancer immunity cycle

Priming“Start Ignition”

Supply co-stimulatory signals“Step on Gas”

Remove inhibitors“Release Brakes”

1

2

3

Co-stimulators

Block co-inhibitors

Chen and Mellman, Immunity, 2013


Lack of compelling monotherapy efficacy in an ‘immune resistant’ or ‘cold’ strain of the MC38 colon cancer model, MC38’R’

8 12 16 20 24 28 32 36 400

500

1000

1500

2000

Anti-PD1 antibody

Day post implantTu

mor

vol

ume

(mm

3 )10 14 18 22 26 30 34 38 42

0

500

1000

1500

2000OX40L mRNA in N1GL

Days post implant

Tum

or v

olum

e (m

m3 )

5 mg/kg 2x/week IP dosing2.5 g Q7D ITu dosing


Robust combination efficacy in MC38’R’ model

• 5 g total mRNA, ITu, Q7D

12 16 20 24 28 32 36 400

500

1000

1500

2000

IL23 with miR122(5g)

Day post implant

Tum

or v

olum

e (m

m3 )

0/12 CRs;8 escapers

12 16 20 24 28 32 36 400

500

1000

1500

2000

IL23_miR122 :IL36_miR122(2.5 g each)

Day post implant

Tum

or v

olum

e (m

m3 )

3/12 CRs;6 escapers

12 16 20 24 28 32 36 400

500

1000

1500

2000

IL23_miR122 : IL36_miR122 :OX40L_miR122

(1.7 g each)

Day post implant

Tum

or v

olum

e (m

m3 )

3/12 CRs;4 escapers

Study performed at Onkaido Therapeutics

Cytokine A mRNA only 5ugCytokine A mRNA 2.5ug +Cytokine B mRNA 2.5ug

Non-translated control mRNA and Cytokine B alone all mice progressed by day 26 (data not shown)

Cytokine A mRNA 1.7ug +Cytokine B mRNA 1.7ug +OX40L mRNA 1.7ug


Summary and Conclusions

• Feasibility and potential of direct intratumoral injection of mRNA encoding immunostimulatory proteins

• Combinations feasible with relative ease

• Significant contribution from lipid to toxicity and tolerability

• Favorable impact of novel, less toxic cationic lipids

• Overcoming resistance to anti-PD1 therapy


• John Zielenski• Susannah Hewitt• Dyane Bailey• Russell Karp• Kana Ichikawa• Ameya Apte• Ailin Bai• Bo Ying• Josh Frederick

• Maja Sedic• Kerry Benenato• Alex Bulychev• Sarah Peterson• Ellalahewage Kumarasinghe• Staci Sabnis • Tim Salerno

Acknowledgments

And a load of other people probably not mentioned by name here

Healthcare

Improved outcomes after intratumoral administration of immunostimulatory mRNA in a novel cationic lipid