Agonist by Means of an Antibody-Drug Conjugate Induces ...€¦ · Marc Damelin, Ph.D. Innate...

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

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Holistic Approach to Build the Optimal STING ADC

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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

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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

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Benchmark1C3176C3165

IRF3

Act

ivat

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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

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• 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

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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

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AntigenAntigenAntigen

FcγR

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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

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Vehicle

Targeted ADC

Control ADC

Free payload

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- 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

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ADC-20 nM

PBMCs + Cancer cells

Control

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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

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Sustained Tumor Regressions Induced by a Single Administration of STING ADC

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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])

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10 / 10 Tumor Free

Target-Dependent Immune Cell Infiltration and Cytokine Induction in Tumors

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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

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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)

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Luminex assay

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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

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0 7 14 21 28 35 42 49 56 63 70 77 840

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Days on Study

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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

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Sustained Tumor Regressions After a Single Dose in a Syngeneic Model

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6/9 Tumor-free

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Dose: mAb / payload (mg/kg)

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ody

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Immunological Memory Induced by STING Agonist ADC

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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)

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Dose: mAb/ payload mg/kg)

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Re-challenge

Conclusions

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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

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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