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Tumor Targeting of a STING Agonist by Means of an
Antibody-Drug Conjugate Induces Potent Anti-Tumor
Immune Responses
Marc Damelin, Ph.D.
Innate Immunity Stimulating Therapies Summit Digital Event – July 22, 2020
Innovative and Highly Differentiated ADC Technologies and Platforms
2DAR = Drug-to-antibody ratioSTING = Stimulator of Interferon Genes
Efficacy without severe neutropenia,
neuropathy, or ocular toxicity
DolaLock Dolaflexin
Improved therapeutic index vs. other
platforms
Dolasynthen
Homogenous & Customizable Platform
Immunosynthen
Systemic administration with targeted immuno-stimulatory effect
• Controlled bystander effect
• Selectively toxic to rapidly dividing cells
• Not a PgP substrate
• Induces immunogenic cell death
• DolaLock payload
• Polymer scaffold
• DAR ~10-12
• Excellent drug like properties
• DolaLock payload
• Synthetic scaffold
• Site-specific
• Precise DAR (2-24)
• Novel STING agonist
• Complete regression with one dose in multiple preclinical models
• Limited effect on systemic cytokines
• Preclinical evidence that STING activation induces prolonged anti-tumor activity and generates immune memory• Other agonists, including TLR7/8, have not shown similar activity in reported studies
• STING activation is more specific to potent Type I interferon gene activation, while TLR activation is associated with general inflammation
• Emerging clinical evidence that STING agonists (intratumoral injection) activate the pathway and do not have significant tolerability concerns
• STING agonists are highly compatible with bioconjugation through the platform technology as they have favorable physicochemical properties
• Oligonucleotides are less compatible (i.e. TLR9, RIG-1)
• Mersana has focused on non-CDN agonists
Why We Invested in STING over other Innate Immunity Pathways
3
ADCs are suited to overcome limitations of free agonist (intratumoral or IV)
– Targeted delivery reduces toxicity liabilities • Minimize toxicity to T and B cells by selective targeting of ADCs (T cell intrinsic function)• Minimize systemic inflammation
– Improved pharmacokinetics
– Accessibility to metastatic sites
– No restriction on tumor type, location or size
Strong Rationale for a STING ADC Approach
4
Holistic Approach to Build the Optimal STING ADC
5
1. Platform• Payload• Linker• Scaffold• Drug-to-Antibody Ratio (DAR)
2. Target and Antibody• Immune cell antigens• Tumor cell antigens• Tumor-associated antigens
Payload molecule
Drug load per scaffold
Charge balance
Aqueous solubilityBioconjugationAntibody
Evaluation included:• Analytical• In vitro characterization• In vivo characterization• Developability
• The target cell is not necessarily a tumor cell– Potential for new mechanisms for payload delivery– Implications for choice of targets and antibodies
• Optimal payload requirements are not known– Potency– Membrane permeability & efflux properties– Metabolism rate (once released)
• Special considerations for I-O in vivo studies– Xenograft models are grown in immune-compromised mice– Syngeneic models are not compatible with certain targets and antibodies
Immunostimulatory ADC Platform Development Cannot Be Based Solely on Cytotoxic ADC Experience
6
Platform Development
• Identified novel compounds representing multiple series– Compounds have a range of biological activity & diverse physicochemical properties
• Leveraged structure-based drug design (SBDD) and crystallography– Crystal structure solved with novel ligand bound to STING
• Filed IP
Novel STING Agonists Designed for ADCs
*Ligand has been removed from structure 8
0.01 1 100 100000
10000
20000
30000
40000
Log Drug Conc. (nM)
Benchmark1C3176C3165
IRF3
Act
ivat
ion
Examples ofnovelMersanapayloads
Payload 2
Payload 1
Leveraging Our ADC Expertise and Proprietary Technologies
10.0 12.5 15.0 17.5 20.0 2
ADCmAb
Aggregation
12.5 15.0 17.5 20.0 22
ADC (mAb #1)ADC (mAb #2)
Optimization
Aggregation in Initial STING ADCNo Aggregation after Optimization
(STING ADCs from 2 different mAbs)
Modular Synthemer Approach Enables ADC Optimization
Payload molecule
Drug load per scaffold
Charge balance
Aqueous solubilityBioconjugationAntibody
Summary of Exploratory Toxicology in NHP
10
• Evaluated ADCs based on 3 antibodies• Dosed up to 9 mg/kg antibody (~0.3 mg/kg STING agonist)• Repeat-dose and single-dose cohorts• Clinical observations
• All animals appeared normal throughout study• No changes in body temperature• No mortality or unscheduled euthanasia
• Toxicokinetics• High exposure after both administrations; dose dependent; overall profile similar to non-STING ADCs• ADC highly stable in circulation; minimal free payload in plasma
• Serum Cytokines• Transient, modest elevation of 5 cytokines out of 24 tested; similar to results in mouse
• No adverse changes in hematology or clinical chemistry• No adverse findings in histopathology
Targets and Antibodies
Comprehensive Approach to Target Validation and Selection
12Image based on DOI: 10.5772/intechopen.72648
Cell Types in the Tumor Microenvironment
Macrophages
Dendritic cellsTumor Cell and Tumor-associatedAntigens
STING Agonist ADCs with Complementary Therapeutic Rationales Based on Antigens and Target Cells
13
PotentialActivation
Activation
Delivery to 2 Cell Types
Target Category Rationale
Immune Cell • Direct activation of immune cells
Tumor Cell• Delivery to tumor and immune cells• Tumor-targeted delivery
Tumor-Associated• Proximity of antigen to immune cells• Tumor-targeted delivery
0.01 0.1 1 10 100 10000
10000
20000
30000
40000
Agonist [nM]
Luci
fera
se A
ctiv
ity
0.01 0.1 1 10 100 10000
50000
100000
150000
Agonist [nM]
Luci
fera
se A
ctiv
ity
Targeted ADC Targeted ADC – Fc MutantControl ADCFree Payload
AntigenAntigenAntigen
FcγR
Antibody variantsTargetedAntigen and
FcγR binding
ControlNo antigen binding
Fc mutantNo FcγR binding
X X
Co-culture assay
Immune CellIRF3 reporter(Luciferase)
Cancer Cell
Recombinant antigen assay
Immune CellIRF3 reporter(Luciferase)
Antigen-coated plate
Tumor-Targeted ADC Activates STING in Immune Cells
FcγRFcγR
Legend
Tumor-Targeted STING Agonist ADC Induces Killing of Cancer Cells by PBMCs
15
Vehicle
Targeted ADC
Control ADC
Free payload
1
10100
100
10
100
10
Agonist [nM]
- Cancer cells have stable expression of a red fluorescent protein in the nucleus
- These cancer cells have minimal STING activity
PBMCs
Co-culture:Cancer cells & PBMCs
100-Fold Increased Potency of ADC over Free Agonist
T Cells are Dispensable for Cancer Cell KillingSupports the hypothesized mechanism of action
PBMCs
CD3
CD
14
CD3+50%
Enriched Monocytes (CD14+CD16+)
CD3+4%
CD16-depleted Monocytes(CD14+CD16-)
CD3+2%
Control
Payload-4 nM
ADC-4 nM
Payload-20 nM
ADC-20 nM
PBMCs + Cancer cells
Control
Payload-4 nM
ADC-4 nM
Payload-20 nM
ADC-20 nM
Enriched Monocytes + Cancer cells Control
Payload-4 nM
ADC-4 nM
Payload-20 nM
ADC-20 nM
CD16-depleted Monocytes+ Cancer cells
16
Sustained Tumor Regressions Induced by a Single Administration of STING ADC
0 7 14 21 28 35 42 49 56 63-20
-10
0
10
20Body Weights
Days on Study
Bod
y W
eigh
t Cha
nge
(%)
Mea
n +/
- SEM0 7 14 21 28 35 42 49 56 63
0
250
500
750
1000Tumor Growth Inhibition
Days on study
Mea
n Tu
mor
Vol
ume
(mm
3 )±SE
M
diABZI I.V. agonist (0 / 5 mg/kg)
Control ADC (1 / 0.04 mg/kg)Control ADC (3 / 0.12 mg/kg)
Targeted ADC (1 / 0.03 mg/kg)Targeted ADC (3 / 0.09 mg/kg)
Unconjugated Antibody (3 / 0 mg/kg)
• All groups dosed IV• Doses in (mAb [mg/kg] / Payload [mg/kg])
17
Vehicle
Legend
10 / 10 Tumor Free
Target-Dependent Immune Cell Infiltration and Cytokine Induction in Tumors
18
Murine cytokine expression(qPCR on FFPE samples)
• ADC single dose• Tumors harvested 12 or 72 hrs post dose
Vehicle
12 hrs.
72 hrs.
Targeted ADC Control ADC
CD45 Immunohistochemistry
0
5
10
15
20
25C C 0
0
0.4
0.8
1.2
1.6
0
2
4
6
12 H 12 H 12 H 72 H72 H72 H
VehicleTargeted ADCControl ADC
mCXCL10 mIL-6mIFNβ
Dramatically Lower Induction of Serum Cytokines in Mice by STING ADC Compared to Free STING Agonist
CXCL10 IL-6 CXCL1 MIG (CXCL9)
RANTES (CCL5)
pg/m
L
0 20 40 60 800
1000
2000
3000
4000
Hours
0 20 40 60 800
5000
10000
15000
20000
25000
Hours0 20 40 60 80
0
2000
4000
6000
Hours0 20 40 60 80
0
2000
4000
6000
Hours
0 20 40 60 800
5000
10000
15000
Hours
0 20 40 60 800
5000
10000
15000
Hours
0 20 40 60 800
1000
2000
3000
4000
5000
Hours0 20 40 60 80
0
50
100
150
200
Hours
0 20 40 60 800
20
40
60
80
Hours
0 20 40 60 800
200
400
600
Hours0 20 40 60 80
0
200
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600
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1000
Hours0 20 40 60 80
0
100
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Hours
0 20 40 60 800
500
1000
1500
Hours
0 20 40 60 800
500
1000
1500
2000
Hours
0 20 40 60 800
20
40
60
80
Hours0 20 40 60 80
0
5
10
15
Hours
MIP-1a (CCL3)TNF-α IFNγ
pg/m
L
Luminex assay
0 5 10 1510
100
1000
10000
100000
Plasma Concentration
Time (Days)
Con
cent
ratio
n [n
g/m
l]M
ean
+/- S
D
Total AntibodyConjugated Drug
STING ADC Yields Extended Exposure
Vehicle control
Targeted ADC (0.09 mg/kg payload = dose for complete tumor regression)
Control ADC (0.09 mg/kg payload)
STING diABZI I.V. agonist (5 mg/kg payload = maximum tolerated dose; 37% tumor growth inhibition) 19
Another Target and Tumor Model:STING Agonist ADC Inhibits Tumor Growth After a Single Dose
20
0 7 14 21 28 35 42 49 56 63 70 77 840
300
600
900
1200Tumor Growth Inhibition
Days on Study
Mea
n Tu
mor
Vol
ume
(mm
3 )±SE
M
Control ADC (3 / 0.1 mg/kg)Targeted ADC (3 / 0.1 mg/kg)• All groups dosed IV• Doses in (mAb [mg/kg] / Payload [mg/kg])
Vehicle
Legend
0 7 14 21 28 35 42 49 56 63 70 77 84-20
-10
0
10
20
Body Weights
Days on study
Bod
y W
eigh
t Cha
nge
(%)
Mea
n±
SEM
Sustained Tumor Regressions After a Single Dose in a Syngeneic Model
0 7 14 21 28 350
500
1000
1500Tumor Growth - Group Means
Days on study
Mea
n Tu
mor
Vol
ume
(mm
3 )±SE
M
21
6/9 Tumor-free
2/10Tumor-free
Dose: mAb / payload (mg/kg)
0 7 14 21 28 35-20
-10
0
10
20Body Weight
Days on studyB
ody
Wei
ght C
hang
e (%
)M
ean±
SEM
0 7 14 21 28 35 42 490
500
1000
1500
2000
Targeted ADC (0.88 / 0.04 mg/kg)
Days on Study
Tum
or V
olum
e (m
m3 )
0 7 14 21 28 35 42 490
500
1000
1500
2000
Control ADC (1.09 / 0.04 mg/kg)
Days on studyTu
mor
Vol
ume
(mm
3 )0 7 14 21 28 35 42 49
0
500
1000
1500
2000Vehicle
Days on study
Tum
or V
olum
e (m
m3 )
Immunological Memory Induced by STING Agonist ADC
220 7 14 21 28
0
400
800
1200
1600Non-targeted tumor
Days post re-challenge
Mea
n Tu
mor
Vol
ume
(mm
3 )±SE
M UntreatedControls
6/6Tumors
• Tumor free mice re-implanted with:• Original targeted tumor on the opposite flank
(blue), and• Non-targeted tumor on the other flank (red).
• Untreated age matched mice also implanted as a control (black line).
Efficacy Study Rechallenge Study (Dual Flank)
0 7 14 21 280
50
100
150
200
250
Targeted tumor
Days post re-challenge
Mea
n Tu
mor
Vol
ume
(mm
3 )±SE
M
UntreatedControls
6/6Tumor-free
Dose: mAb/ payload mg/kg)
0 7 14 21 28 35 42 490
500
1000
1500
2000
Targeted ADC (0.88 / 0.04 mg/kg)
Days on Study
Tum
or V
olum
e (m
m3 )
6/9 Tumor-free
Re-challenge
Conclusions
23
1. STING agonist ADC platform• Novel agonist payload optimized for ADC
• Linker & scaffold designed to maximize therapeutic index
• Well-tolerated in non-human primates
2. In vivo activity in multiple targets, tumor models and mouse strains
3. Differentiation from IV agonist: activity and tolerability
4. Immunological memory
5. On track to nominate 1st Development Candidate in 2020
Mersana’s STING ADC Research Team
24
Discovery ChemistryJeremy DuvallJosh ThomasKeith BentleyBrian JonesEoin KelleherLiping YangRadha IyengarDorin Toader
BiologyRaghida Bukhalid Naniye CetinbasChen-Ni ChinScott CollinsTimothy EitasWinnie LeeTravis MonnellMarina ProtopopovaLiuLiang Qin Kelly SlocumMarc Damelin
Analytical & BioanalyticalKenneth AvocetienStephen BradleyTyler CarterSusan ClardyMark NazzaroElena Ter-OvanseyanLing XuJeffrey ZuritaDavid Lee
Translational MedicinePamela ShawRebecca Mosher
Chief Science & Technology OfficerTimothy B. Lowinger
BioconjugationKalli CatcottBingfan DuCheri StevensonAlex UttardAnouk DirksenDorin Toader