a first-in-class decoy oligonucleotide ......asidna is a first-in-class product in dna damage...
TRANSCRIPT
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
1st cycle 2nd cycle 3rd cycle 4th cycle0
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)
MDA-MB-231 (TNBC HR proficient tumor cells) xenograft in mice
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
no rescueafter 1st cycle
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
no rescueafter 1st cycle
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
no rescueafter 1st cycle
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
1st cycle 2nd cycle 3rd cycle 4th cycle0
20
40
60
80
100
Talazoparib AsiDNA+Talazoparib
Surv
ival (
%/N
T)
1st cycle 2nd cycle 3rd cycle 4th cycle0
20
40
60
80
100
120
140
Olaparib Talazoparib
AsiDNA+Olaparib AsiDNA+Talazoparib
Surv
ival (
%/N
T)
Source: Onxeo, data on file
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
MDA-MB-231 (TNBC HR proficient tumor cells) xenograft in mice
Source: Onxeo, data on file
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
Source: Onxeo, data on file
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
Pearson r: 0.55P<0.05
Pearson r: 0.62P<0.05
Pearson r: 0.6P<0.05
Pearson r: 0.6P<0.05
Pearson r: 0.54P<0.05
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