A FIRST-IN-CLASS DECOY OLIGONUCLEOTIDE ......AsiDNA is a first-in-class product in DNA Damage...

AsiDNA™A FIRST-IN-CLASS

DECOY OLIGONUCLEOTIDETARGETING DNA REPAIR

PATHWAYS TO KILL TUMOR CELLS

2ndAnnual DNA Damage Response Therapeutics Summit

January 29-31 2019, Boston, MA

AsiDNA is a registered trademark of Onxeo

Françoise BonoChief Scientific Officer

2

Recruitment of DNA repair proteins to the sites of

genomic damage is one of the early events

in DNA repair

AsiDNA™ is a decoy oligonucleotide that mimics double-strand DNA breaks,

redirecting repair enzymes away from the real sites of

DNA damage

Tumoral cells (deficient cell cycle control)

keep dividing with damaged DNA leading to

DNA fragmentationand cell death by mitotic

catastrophe

Healthy cells (proficient cell cycle

control) stop dividing and stop transcription until

AsiDNA™ clearance

AsiDNA™ - A mechanism of action unlike any other:DNA repair inhibition through a decoy strategy

Global gene expression by principal component analysis (PCA)

Treatment with AsiDNA™ dramaticallymodifies genes expression profile in cancer

cells but not in healthy cells

January 2019

3AsiDNA™ is a first-in-class product in DNA Damage Response

A synthetic cholesterol-oligonucleotide conjugate forming an intramolecular hairpin 32-base pair double helix

Robust GMP manufacturing process scaled up to 1,7kg, long shelf stability

No formulation needed to achieve sufficient plasmatic and tumor exposure after intravenous administration confirmed in vivo as well as in DRIIV-1 trial

5’

3’

3’

5’

Genomic DNA length optimized to bind and activate DNA-PK and PARP signaling enzymes

Phosphorothioate substitutions at the 5’ and 3’ ends to prevent degradation1

Double-stranded 32 bp DNA is tethered with a loop to prevent disassociation1

Sequence not specific, chosen to be non-homologous and not immunogenic (CpG-free)

Molecular weight : 22’359.2 daltons

Active 32 bp DNA duplex

Cholesterol

Loop

1. Quanz M, et al. PLoS ONE. 2009 4(7), doi: 10.1371/journal.pone.00062982. Berthault N, et al. Cancer Gene Therapy (2011), 1-12, doi: 10.1038/cgt.2011.3

Efficient tumoral and nuclear uptake of the DNA is mediated via a covalently linked cholesterol molecule2

January 2019

4AsiDNA™ triggers an artificial DNA damage signaling

AsiDNA™ traps and hyper-activates multiple key DDR enzymes from multiple pathways

Pool down with biotinylated AsiDNATM

10 Gy irradiation

DNA-PK auto-

phosphorylation

AsiDNA™

PARPauto-parylation

γ-H2AX phosphorylation

AsiDNATMNT

53BP1phosphorylation

MO59 K Glioma cells

Adapted from Targeting the DNA Damage

Response for Anti-Cancer Therapy, John Pollard, Nicola Curtin Chap. 14. 2018

Targets trapped by AsiDNA™ belong to multiple pathways

Ku70DNA-PK

PARP1XRCC1

PCNA

PCNAPCNA

PCNA

PCNA

HR

NHEJ

a-NHEJ

BERSSBR

MMR NER

FAFEN1APE1

POLB/D/ELIG3

MSH2/MSH6EXO1

RPA/RFCPOLD/E

FA core complexFANCMFAN1

XPA/G/XPC-Rad23BERCC2/3/4/5/6/8

RPA1/2TFIIH

POLD/ELIG1

CCL1CDK7

CETN2DDB1/2

ERCC1/XPF

CtIPMRN

RPA1/2RAD51/52/54

XRCC3BRCA1/2LIG1/3

XRCC4XLF

ArtemisPOLu/D

PLKLIG4

53BP1CtIPMRNLIG3

January 2019

5

Jdey W et al, submitted

AsiDNA™ leads to cancer cell death and does not induce resistance

Treatment with AsiDNA™ reduces cancer cells survival (1-30µM IC50), while healthy cells are spared

In contrast to targeted therapies (eg PARPi), repeated treatment with AsiDNA™ leads to sensitization to AsiDNA™ and does not generate resistance

Suggests a compelling opportunity as a maintenance therapy

BC227 – TNBC HR BRCA1 mutated

NCI-H446

1st cycle 2nd cycle 3rd cycle 4th cycle0

20

40

60

80

100

120AsiDNA Talazoparib

Surv

ival (%

/NT)

*

NCI-H446 – SCLC

BC227


20

40

60

80

100

120

AsiDNA Talazoparib Olaparib

Surv

ival (%

/NT)

January 2019

6

MDA-MB-231 (TNBC HR proficient tumor cells) xenograft in mice

Treatment

1st cycleTreatment

2nd cycle

Treatment

3rd cycleRelease Release Release

AsiDNA™

1 cycle = five consecutive days (5 mg/injection/day) followed by 17 days of rest

5 mg/day ip

Jdey W et al, submitted

Repeated treatment with AsiDNA™ induces auto-sensitization in vivo

January 2019

7AsiDNA™ autosensitization properties : mechanistic considerations

The unusual evolution of the tumor cells under AsiDNA™ treatment toward increased sensitivity, could reveal :

The selection of a preexisting population more sensitive to AsiDNA™

The evolution of all the treated cells in a new state in absence of any selection process

The accumulation of cell damage or genetic instability at each cycle of AsiDNA™ treatment due to unrepaired or misrepaired chromosomes that will require a constitutive repair activity which has been lost during treatment

January 2019

8Combination of AsiDNA™ with carboplatin generates synergies and prevents resistance to carboplatin

January 2019

AsiDNA™ in combination with carboplatin

NCI-H446 Carboplatine

1st c

yle

2nd c

ycle

3rd c

ycle

4th c

ycle

5th c

ycle

0

20

40

60

80

100

120NT

Carboplatine

AsiDNA+Carboplatine

Surv

ival (

%/N

T)

[AsiDNA™]: 2,5µM [carboplatin]: 2,5µM

- SCLC

Carboplatincarboplatin

Source: Onxeo, data on file

Prevents the occurrence of resistance to carboplatinShows synergistic efficacy

0.0 0.5 1.00

20

40

60

80

100

120

Carbo Carbo + AsiDNA 5µM

* * **

CI=0.5

CI=0.29

[Carbo, µM]

Surv

ival (%

/NT)

MDA-MB-231 -TNBC

9Combination of AsiDNA™ with carboplatin shows synergy in resistant TNBC

0

100

200

300

400

500

600

700

800

900

1000

0 10 20 30 40 50 60 70 80 90 100

Mean

volu

mes (m

m3)

Time after treatment (days)


Group I : vehicle (Nacl 0.9%)

Group II : AsiDNA (IP)

Group III : carboplatin (IP)

Group IV : AsiDNA (IP) + carboplatin (IP)

Endpoint1500 mm3

d - XX graft

Week 7Week 4Week 1

Source: Onxeo, data on fileJanuary 2019

Treatment Median survival(days)

Vehicle NaCl 0,9% 15x IP (n 6) 77

Carboplatin 3x 50 mg/kg IP (n 8) 88

AsiDNA 15x 5 mg IP (n 8) 128

AsiDNA 15x 5mg IP + carboplatin 3x 50mg/kg IP (n 10)

175

10

Data support increased efficacy of the combination of AsiDNA™ with DNA-damaging agents in multiple in vivo models

Tumor model (cell lines, primary tumors)

Treatment in combination Route of administration Reference

Breast cancer (BC227, BC173, MDA-MB468, MDA-MB231) AsiDNA™ standalone

Intratumoral + Peritumoral Subcutaneous Intraperitoneal (MDA-MB231) Data on file

Breast cancer (MDAMB231, BC227) AsiDNA™ + carboplatin Intraperitoneal Data on file

Glioblastoma AsiDNA™ + Radiotherapy Intratumoral Coquery et al, 2012

Cutaneous melanoma AsiDNA™ + RadiotherapySubcutaneous / PeritumoralIntratumoral + subcutaneous / Peritumoral

Schlegel et al, 2012Biau et al, 2014

Colorectal cancer AsiDNA™ + RFA (hyperthermia) Intratumoral + subcutaneous Devun et al, 2014

Colorectal Liver metastasis (HT29) AsiDNA™ + 5FU + oxaliplatin Intraperitoneal Herath et al, 2016

HCC (HepG2) AsiDNA™ + doxorubicin Intraperitoneal Herath et al, 2016

Head & neck (Hep2) AsiDNA™ + Radiotherapy Intratumoral Quanz et al, 2009

Head & Neck (Hep2) AsiDNA™ + carboplatin Intraperitoneal Data on file

Lung cancer (TC-1) AsiDNA™ + carboplatin or cisplatin Intraperitoneal Data on file

HCC (VX2 rabbit) AsiDNA™ + TACE (Doxorubicin) Transarterial Herath et al, 2016

January 2019

11AsiDNA™ in combination with PARPi: synergy in TNBC and SCLC

BC227

1st cycle 2nd cycle0

20

40

60

80

100

120NT

AsiDNA

Talazoparib

AsiDNA+Talazoparib

no rescueafter 1st cycle

Surv

ival (

%/N

T)

NCI-H446

1st cyle 2nd cycle0

20

40

60

80

100

120NT

AsiDNA

Talazoparib

AsiDNA+Talazoparib


Surv

ival (

%/N

T)

TNBC HR deficientBC227

Small cell lung cancerNCI-H446BC227

1st cycle 2nd cycle0

20

40

60

80

100

120NT

AsiDNA

Talazoparib

AsiDNA+Talazoparib


Surv

ival (

%/N

T)

1 week recovery

No recovery after a wash out of 7 days

NCI-H446

1st cyle 2nd cycle0

20

40

60

80

100

120NT

AsiDNA

Talazoparib

AsiDNA+Talazoparib


Surv

ival (

%/N

T)

1 week recovery

No recovery after a wash out of 7 days

100 nM5µM

AsiDNA™ in combination with PARPi

Shows the same efficacy in both HR deficient and proficient cancer cells; combining AsiDNA™ with PARPi may expand PARPi’s use to HR-proficient tumors (data not shown)

Leads to a rapid and complete inhibition of cancer cell survival, which is not observed with PARPi alone (synergistic effect)

January 2019

12AsiDNA™ prevents resistance to PARPi

AsiDNA™ in combination with PARPi

Prevents at low doses the occurrence of resistance in several cancer cell lines

TNBC HR deficient (BC227) SCLC (NCI-H446)

Olaparib 5 µM - Talazoparib 100nM - AsiDNA™ 1µMTalazoparib 100nM – AsiDNA™ 1µM


20

40

60

80

100

Talazoparib AsiDNA+Talazoparib

Surv

ival (

%/N

T)


20

40

60

80

100

120

140

Olaparib Talazoparib

AsiDNA+Olaparib AsiDNA+Talazoparib

Surv

ival (

%/N

T)


January 2019

13Combination of AsiDNA™ with PARPi demonstrates synergy in HR proficient TNBC

AsiDNA™ in combination with PARPi shows high efficacy in vivo in cancer cells non-sensitive to PARPi

Opportunity to expand PARPi indications to HR proficient tumors

NT olaparib AsiDNA™ AsiDNA™ + olaparib

Complete Response = 0/6 Complete Response = 2/8 Complete Response = 4/8 Complete Response = 5/7 0% 25 % 50% 71%

January 2019



14Class-effect of the combination of AsiDNA™ with PARPi’s supportedby extensive preclinical testing

Tumor model (cell lines, primary tumors)

Treatment in combination Observed effect Reference

TNBC (MDA-MB-231xenograft mice model)

AsiDNA™ + olaparib Complete response more than doubled Data on file

Resistant PDX ovarian model AsiDNA™ + olaparib Synergy delaying tumor growth Data on file

TNBC HR deficient (BC 227) AsiDNA™ + olaparib or + talazoparib Resistance is prevented by co-treatment Data on file

Ovarian cancer AsiDNA™ + niraparib Resistance is prevented by co-treatment Data on file

SCLC (NCI-H446) AsiDNA™ + talazoparib Resistance is prevented by co-treatment Data on file

TNBC HR deficient (BC 227) AsiDNA™ + talazoparib Rapid and complete inhibition of cancer cell survival Data on file

SCLC (NCI-H446) AsiDNA™ + talazoparib Rapid and complete inhibition of cancer cell survival Data on file

TNBC HR proficient (MDAMB231) AsiDNA™ + talazoparib Synergistic antitumor effect Data on file

TNBC HR proficient (MDAMB231) AsiDNA™ + niraparib Synergistic antitumor effect Data on file

TNBC HR proficient (MDAMB231) AsiDNA™ + olaparib Synergistic antitumor effect Data on file

Both synergy and abrogation of resistance to treatment occurwith all tested PARPi and regardless of the tumor type,

opening new opportunities for clinical applications January 2019

15AsiDNA™ differentiating features validated in both preclinical and clinical studies*

*Data published, submitted for publication or on file at Onxeo

Preclinical package confirming unique properties of AsiDNA™ alone and in combination*

Favorable safety and PK profile via intravenous administration

Dose proportionality either in CMax and AUC

No drug-related serious adverse events, no MTD

PK/PD relationship demonstrated in pharmacological models

Target engagement in tumors as measured by DNA-PK activation (gH2AX) is related to plasmatic exposure

January 2019

Repeated treatment with AsiDNA™ leads to sensitization to AsiDNA™ and does not generate resistance

Resistance to PARPi treatment is prevented by co-treatment with AsiDNA™

Strong synergy of AsiDNA™ in combination with PARPi

Strong synergy of AsiDNA™ in combination with carboplatin

Efficacy and safety demonstrated in man via local administration

59% Overall RR in metastatic melanoma (AsiDNA™ + radiotherapy) via IT route

16

PHASE I

Open-label, 3+3 dose escalation

2 European countries: FR, BE

5 centers: Paris(2),Toulouse, Lyon, Brussels

Study coordinator: Pr. C. Le Tourneau (Institut Curie)

DSMB

OBJECTIVES To determine dose-limiting toxicities (DLTs) and the maximum tolerated dose (MTD)

To evaluate the pharmacokinetics/pharmacodynamics (PK/PD) effects of AsiDNA™ based on biomarkers of activity in blood and in tumor tissues

APRIL 2018FIRST PATIENT DOSED

TREATMENT SCHEDULE

Dose Level 1

Dose Level 2

Dose Level 3

Dose Level 4Dose Level 5

Dose Level 6

200 mg

400 mg

600 mg

900 mg

1300 mg1800 mg

3 patients

4 patients

3 patients

H1 2019 END OF STUDY

1-hour IV infusion

DAY 1 21 2 3 8 15

CYCLE 1CYCLE 2 and beyond : once a week

DRIIV-1 study: evaluating AsiDNA™ via IV administration DNA Repair Inhibitor administered IntraVenously in patients with advanced solid tumors (in patients failing previous anticancer therapies)

3 patients

ACTIVE DOSES FOUND

Q4 2018 BIOACTIVITY

6 patients

November 5, 2018

Compelling safety & activity observed as early as dose level 2

ongoing

January 2019

17

AsiDNATM Safety at 200mg (DL1), 400 mg (DL2) & 600 mg (DL3) (n = 10 patients / infusions = 112)

No drug-related serious adverse events

No dose-limiting toxicity

No cumulative relevant safety

Only grade 1 & 2 drug-related adverse events

Maximum tolerated dose not reached

Drug-related AE

Grade 1Grade 2

DRIIV-1 study: interim results (DL1 to DL3 completed)Safety/PK

AsiDNA™ pharmacokineticsDose proportionality across the 3 dose levels

Cmax and AUC are increased proportionally and consistently with dosing


Dose(mg)

Cmax(µg/mL)

AUC (t0-tlast) (µg.h/mL)

200 41,8 110,9

400 76,9 161,6

600 129,4 247,1

January 2019

18DRIIV-1: interim results (DL1 to DL3 completed)Pharmacodynamics - Proof-of-Mechanism in man

Analysis of the first 3 dose levels out of 6 planned

10 patients

4 biopsies available at baseline and end of cycle 2

Target engagement confirmed by significant increaseof activity biomarkers* from dose level 2 and 3

= Proof-of-Mechanism of AsiDNA™ in man

Decrease or stabilization of tumor proliferationrate from dose level 2 and 3

Activity

Robust target engagement in patients’ tumors demonstrates AsiDNA™ activity via IV route

Tumor status

Activity biomarkers

Significant increase after treatment with AsiDNA™

Major increase after treatment with AsiDNA™

DL2 : 400mg gH2AX pHSP90

Patient 0106

Patient 0109

DL3 : 600mg gH2AX pHSP90

Patient 0202

Patient 0301

Activity biomarkers

DL2 : 400mg KI67

Patient 0106

Patient 0109

DL3 : 600mg KI67

Patient 0202

Patient 0301

Tumor proliferationbiomarker

Tumor proliferationbiomarker

Source: Onxeo, data on file* gH2AX and pHSP90 are two established biomarkers of the activation of DNA-PK, a key DDR enzyme and a major target for AsiDNA™

January 2019

19AsiDNA™ predictive activity biomarkers identification for patient stratification

Trancriptomic analysis

Breast cancer cell lines

Small cell lungcancer cell lines

AsiDNA™ signature

Breast Cancer

Lung Cancer

Top 25% AsiDNAsensitive tumors

DDR genes expressionMostly downregulation

6 genesdownregulation the

most highlycorrelated with

AsiDNA sensitivity

Ranking of cells for their sensitivity to AsiDNA™

BRCA2

-11 -10 -9 -8 -70

20

40

60

80

100

120

Hep3B2.1.7

MDA-MB-157

HuH7

BxPC3

Mia Paca2

HepG2

SNU-423

U2-OS

A549

MDA-MB-436SNU-449

MDA-MB-231

BC227

FaDu

NCI-H446 (adh)

BRCA2 expression level

Surv

ival t

o A

siD

NA

(%

/NT)

M R E 1 1 A

- 1 1 - 1 0 - 9 - 8 - 7 - 6 - 50

2 0

4 0

6 0

8 0

1 0 0

1 2 0

Hep3B2.1.7

MDA-MB-157

HuH7

BxPC3

Mia Paca2

HepG2

SNU-423

U2-OS

A549

MDA-MB-436 SNU-449

MDA-MB-231

BC227

NCI-H446 (adh)

M R E 1 1 A e x p r e s s i o n l e v e l

Su

rv

iv

al

to

A

siD

NA

(

%/

NT

)

XRCC2

-11 -10 -9 -8 -7 -60

20

40

60

80

100

120

Hep3B2.1.7

HuH7

BxPC3

Mia Paca2

HepG2

SNU-423

U2-OS

A549

MDA-MB-436SNU-449

MDA-MB-231

BC227

FaDu

NCI-H446 (adh)

XRCC2 expression level

Surv

ival t

o A

siD

NA

(%

/NT)

NBN

-8 -7 -6 -50

20

40

60

80

100

120

Hep3B2.1.7

MDA-MB-157

HuH7

BxPC3

Mia Paca2

HepG2

SNU-423

U2-OS

A549

MDA-MB-436

MDA-MB-231

BC227

FaDu

NBN expression level

Surv

ival t

o A

siD

NA

(%

/NT)

FANCA

-11 -10 -9 -8 -7 -60

20

40

60

80

100

120

Hep3B2.1.7

MDA-MB-157

HuH7

BxPC3

Mia Paca2

HepG2

SNU-423

U2-OS

A549

MDA-MB-436SNU-449

MDA-MB-231

BC227

NCI-H446 (adh)

FANCA expression level

Surv

ival t

o A

siD

NA

(%

/NT)

Rad54b

-11 -10 -9 -8 -70

20

40

60

80

100

120

Hep3B2.1.7

MDA-MB-157

HuH7

BxPC3

Mia Paca2

HepG2

SNU-423

U2-OS

A549

MDA-MB-436

MDA-MB-231

BC227

FaDu

NCI-H446 (adh)

Rad54B expression level

Surv

ival t

o A

siD

NA

(%

/NT)

Pearson r: 0.75P<0.01






Bioinformatic

Demonstrated significant correlation between the

expression of the 6 genes & sensitivity to AsiDNA™

in cancer cells

Experimental

Predicted resistant Predicted sensitive

0

20

40

60

80

100

120

**

Surv

ival to

AsiD

NA

(%

/NT)

Rankingvalidation at the bench

Ongoingin vivo

validation

January 2019

20AsiDNA™, a highly differentiated molecule with near-term clinical milestones

AsiDNA™ : a unique mechanism of action enabling a new approach to cancer treatment

Exhaustive and robust preclinical package showing product wide potential in combination (PARPi, chemotherapies and radiotherapy)

Biomarker-driven patient selection strategy for AsiDNA™ treatment to be further validated in vivo and in clinic.

Established proof-of-mechanism in man, strong activity, favorable safety, active doses found

January 2019

Ready for expanded clinical development in combination

Final data from DRIIV-1 Phase 1 trialInitiation of a Phase 1b study in combination with platinum-based chemotherapies

Initiation of IND filing in the US

H1 2019

H2 2019

21

• Pr Christophe LE TOURNEAU

• Pr Jean-Pierre DELORD

• Pr Philippe CASSIE

• Dr Nuria KOTEKI

• Jean-Pierre BIZZARI

• Alain HERRERA

• Pr Michel MARTY

TeamScientificCollaborations

AdvisorsClinical

Investigators

• Marie DUTREIX (PhD team)

Acknowledgments

A FIRST-IN-CLASS DECOY OLIGONUCLEOTIDE ......AsiDNA is a first-in-class product in DNA Damage...

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Transcript of A FIRST-IN-CLASS DECOY OLIGONUCLEOTIDE ......AsiDNA is a first-in-class product in DNA Damage...