December 2017

1

Combining Bi-specific Antibodies and

Oncolytic Virus Therapy

Shautong Song, MD. Ph.D.

2

Company Overview

Product Technology IndicationResearch IND

EnablingIn vitro In vivo

TEA-OV Candidates

IKT-901 FAP-TEA-VV Solid tumors

IKT-902 EpCAM-TEA-VV Solid tumors

IKT-903 EphA2-TEA-VV Solid tumors

IKT-904 HER2-TEA-VV Solid tumors

IKT-905 GPC3-TEA-VV Solid tumors

IKT-906 GD2-TEA-VV Solid tumors

IKT-907 FAP-TEA-HSV Solid tumors

VV Expressing Checkpoint Inhibitors

IKT-201 PD-L1-VV Solid tumors

IKT-202 PD-1-VV Solid tumors

IKT-203PD-L1-FAP-TEA-

VVSolid tumors

CAR (Chimeric Antigen Receptor)-T: VV Loaded & Next Gen NK

IKT-701 VV-loaded CAR-T Solid tumors

IKT-702 CD19-CAR-NK Blood tumors

IKT-703 GD2-CAR-NK Solid tumors

• BCM spin-off company at JLABS@Houston

• Technology: Oncolytic virus therapies

• Lead Program: T-cell Engager-Armed oncolytic

virus expressing bi-specific antibodies (TEA-OV)

• Short Timeline to Clinic With Lead Program:

ppIND meeting has been held regarding IKT-901,

which is expected to enter Phase I in Q3 2018

• Manufacturing & Trial Execution Leverages

Leading OV Experts: BCM & UPMC experts for

GMP production and trial execution

• Strong IP Portfolio: Robust and growing portfolio

of filed 4 patents

Company Overview & Key Highlights

1

2

3

3

Oncolytic Viruses: Validated through recent approvals and multiple strategic

partnerships

Licensor/Acquiror Target

BMS PsiOxus

Pfizer Ignite

Merck Viralytics

Merck DNAtrix

Licensor/Acquiror Target

Boehringer ViraTherapeutics

Pfizer Western Oncolytics

Celgene Oncorus

2004 2006 2008 2010 2012 2014 2016+

BioVex first reports efficacy

of OncoVEX in Phase 2

study for melanoma

Amgen’s T-VEC

was approved by

FDA for melanoma

FDA issued

guidance for OV

clinical trials

BioVex was

acquired by Amgen

for $1 billion

ONYX-015 was

approved by CFDA for

head & neck cancer

>7 OV Deals

4

Immuno-oncology: Despite recent advances, limitations remain

Therapy Class Examples%（CR+PR）

LimitationsIKT Approach to Overcome

LimitationsSolid Liquid

Tumor Antigen

mAb

12.2-

32.5%1

• Minimal Immune

Stimulation

• Vaccinia Mediated Oncolysis

• T Cell Stimulation/Recruitment

Bi-specific T cell

Engager88%2

• Side effects

• Suboptimal solid tumor

penetration

• Expression restricted to tumor

• Increase local expression of BiTE

Cytokines 25%3 • Systemic Side effects • Transgene expression limited to tumor

Checkpoint

Inhibitors6-25%4 87%5 • Safety, esp. in combo w/ other

IO• Local expression to avoid systemic effects

DC/CIK Therapy 0%6 • Minimal efficacy to date

• Not an ‘off the shelf’ product• Off the shelf approach

Oncolytic virus 10-26.4%7 • Potency as monotherapy• Armed with I/O potentiation biologics

against multiple unique targets

CAR-T cell therapy Tisagenlecleucel-T 83-95%8

• Safety

• Not an ‘off the shelf’ product

• Efficacy in solid tumors

• CAR-T’s loaded with OV expressing

biologics against solid tumor targets

Sources: 1. McCann et al. Gynecol Oncol. 2012; 127(2); Pietrantonio et al. Oncologist. 2015; 20:1261; Xiong et al. Journal of Clinical Oncology 22:2610.2. http://www.amgen.com/media/news-releases/2015/12/amgen-presents-data-from-three-trials-evaluating-blincyto-blinatumomab-in-acute-lymphoblastic-leukemia-at-ash-2015/3. Buchbinder et al. Journal for Immjunotherapy of Cancer 2016 4:52. https://www.proleukin.com/downloads/24954-Fact-Sheet-and-Questions-for-Your-Doctor-Guide_7-1.pdf4. Kwok et al. Hum Caccin Immunother. 2016:2777; Queirolo et al. Cancer Treat Rev. 2017 19:71; 5.Villasboas et al. F1000 Faculty 2016 5:Rev-7686.Maia et al. Crit Rev Oncol Hematol. 2017 113:2917.Bilsland et al. F100Res 2016 30;58. https://www.fda.gov/downloads/AdvisoryCommittees/CommitteesMeetingMaterials/Drugs/OncologicDrugsAdvisoryCommittee/UCM567385.pdf

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TEA-OV: A novel modality leveraging established IO approaches

Multi-technology approach targeted to novel tumor-specific antigens:

– Oncolytic vaccinia virus expressing bi-specific antibodies

– Targeting FAP, EpCAM, EphA2, HER2, GPC3, and GD2

Superior to other modified VVs:

– Expression and activity does not depend on development of an endogenous

antitumor response (absent/ compromised in many patients)

– Directly engages T cells to kill non-infected tumor cells

– Induces anti-tumor immunity without overdriving innate viral immunity

Addresses limitations of BiTE solid tumor therapy:

– TE continually produced locally within VV-infected tumor translating to

high tumor penetration

– High local TE concentrations reduces the systemic side effects and toxicity

– Viral oncolysis prevents antigen loss variants

Program Highlights

CD3 Ab

Target Ab

Tumor cell

6

FAP-TEA-VV: Overview

• Targeting FAP enhances antitumor activity

− Improve virus spread within tumor tissue by

targeting CAF

− Overcome immune suppressive tumor

environment by targeting TAM

− Enhance bystander killing of FAP+ tumor

• FAP-CAR-T and FAP tumor vaccine have

been evaluated in clinical studies

− The safety of FAP-CAR-T therapy is being

validated in phase 1 clinical study

− A clinical study of FAP-IL2 fusion protein

has been launched

FAP-

TEA-VV

FAP+ CAF

CTL

FAP+ TAM

CTL

Tumor

cell

FAP+ Tumor

CTL

Cancer-Associated Fibroblasts: FAP is

overexpressed in stroma of >90% of epithelial

tumors, and is not expressed in healthy adult tissues

outside of wound repair1

Tumor-Associated Macrophages

Tumor Cells:

A B C

A

B

C

Mechanism of Therapeutic Effect

FAP Expression in Tumor Environment

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PROBLEM: Spread of Virus Limited by Tumor Environment

The antitumor effects of systemic delivery of vaccinia virus is limited by its low virus delivery

efficacy (limited virus access to tumor), and suboptimal virus spread within the tumor tissue

Role of Cancer associated Fibroblasts:

• CAFs produce collagens, laminins, fibronectins, proteoglycans, and hyaluronan, forming a dense

extracellular matrix (ECM) around tumor cells

• CAFs combined with ECM provide physical barriers to oncolytic viruses, forming a shield surrounding

tumor nests

• Oncolytic viruses have limited autonomous motility and can only be spread through cell-to-cell contact

or soluble diffusion across concentration gradients, both of which can be blocked by CAF and ECM

- First, viruses could potentially adhere to any surfaces of CAFs where they are first administered

and then fail to spread since CAFs are rather resistant to virus infection.

- Second, oncolytic VV has a diameter > 200 nm and physically does not fit through the strands of

the ECM, preventing passive diffusion across concentration gradients to reach the tumor.

IKT’s FAP-targeted virus provides an alternative and unique strategy to improve the efficacy of systemic

administration of oncolytic virus, by promoting virus spread and replication within tumors

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Solution: Targeting the ECM through CAFs

Targeting elements of the ECM improves local replication of systemically delivered virus:

• Using matrix-degrading enzymes to remove physical ECM barriers is currently the foremost option as

a study of HSV (a large virus over 100 nm) showed that coinjection of matrix-degrading collagenase

improved viral spread within the tumor

• However, CAFs could produce ECMs continuously and thus the efficacy of matrix-degrading

collagenase could rely on the speed of production and degradation of ECMs

• Despite the limited gains, this provides compelling proof of principle to continually target CAFs as a

means to improve systemic efficacy of OVs

FAP-TEA-VV provides an alternative and unique strategy to improve systemic therapy of OV by:

• Destroying CAFs and removing physical barriers to oncolytic virus in a more permanent manner,

• Allowing small amount of virus to replicate effectively within tumor, and leading to enhanced

antitumor effects

We observed that administration of FAP-TEA-VV in mouse models significantly enhanced viral titer

within tumors, leading to enhanced antitumor effects and the increase of viral titer is correlated with

the destruction of FAP-positive stromal cells

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Fibroblast Activation Protein (FAP)

melanoma

Normal

skin

20X 40X

Hofmeister et al; Cancer Immunol Immunother 2006

10

FAP-TEA-VV: Construction

CD3 AbFAP Ab

FAP+

Tumor cell

1. Recombinant insertion of TE into TK gene

of ∆VGF-VV

2. Plaque purification (YFP+); YFP deletion

(Cre/loxp)

3. Production of FAP-TEA-VV in permissive

cell lines

Production of Recombinant FAP-TEA VV

vTK -L F17R vTK -RFAP-scFv CD3-scFv

FAP Ab

CD3 Ab

FAP+

Tumor cell

CD3+

T-Cell

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Mouse Version FAP-TEA-VV

FAP-TEA-VV

GM-CSF-VV

VGF

ITR 5’TK

GM-SCFPF17R

VGF

ITR

LacZ

P11K

GFPPSEL P11K

LacZ

3’TK

VGF ITR 5’TK

FAP-TEPF17R

VGF

ITR

LacZ

P11K

DsRedPSEL P11K

LacZ

3’TK

EphA2-TEA-VV

A

B C

M 0.5µg 1µg. 2.5 µg

EphA2-CD3 proteinF

AP

-TE

A-

VV

Ep

hA

2-

TE

A-V

V

GF

P-V

V

VGF

ITR 5’TK

EphA2-TEPF17R

VGF

ITR

LacZ

P11K

DsRedPSEL P11K

LacZ

3’TK

0

10

20

30

40

50

60

0 0.1 1 5

GM

-CS

F e

xpre

ssio

n

(pg/m

l)

MOI

B16

GL-261

12

FAP-TEA-VVMedium EphA2-TEA-VV

MOI 1

GFP

48 h

T cells

GL261-

mFAP 24 h 42.2%

61.0%

30.4%

16.4% 0.67%

37.4%

32.8%

EphA2-TEA-VV FAP-TEA-VV

MOI 0.1

28.6%

19.1%

1.67%

FAP-TEA: Enhanced in vitro Killing with T cells

FAP-TEA-VVMedium EphA2-TEA-VV

MOI 1

GFP

48 h

T cells

GL261-

mFAP 24 h 42.2%

61.0%

30.4%

16.4% 0.67%

37.4%

32.8%

EphA2-TEA-VV FAP-TEA-VV

MOI 0.1

28.6%

19.1%

1.67%

GFP Expressing

FAP+ Target Cells

MOI- multiplicity of infection (measure of virus concentration in assay)

T c

ell

Activa

tio

n M

ark

ers

0

2000

4000

6000

8000

Me

diu

m

Ep

hA

2-T

EA

-VV

FA

P-T

EA

-VV

mIF

N-γ

p

g/1

06

T c

ells

MOI 1

MOI 0.1

0

500

1000

1500

2000

Me

diu

m

Ep

hA

2-T

EA

-VV

FA

P-T

EA

-VV

mIL

-2 p

g/ 1

x1

06

T c

ells MOI 1

MOI 0.1

0

2000

4000

6000

8000

Me

diu

m

Ep

hA

2-T

EA

-VV

FA

P-T

EA

-VV

mIF

N-γ

p

g/1

06

T c

ells

MOI 1

MOI 0.1

0

500

1000

1500

2000

Me

diu

m

Ep

hA

2-T

EA

-VV

FA

P-T

EA

-VV

mIL

-2 p

g/ 1

x1

06

T c

ells MOI 1

MOI 0.1

Tumor cells

Murine Construct

A B

13

FAP-TEA-VV: Bystander Killing of Tumor Cells

Grow VV (FAP or Control)

in Tumor cells

Harvest culture medium

(no infected cells present)

Add to FAP+ cells

w/ T cells

Murine Construct

Experiment Design Results

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FAP-TEA-VV: Inhibition of B16 Tumor Growth

FAP-TEA-VV demonstrates robust tumor inhibition in the B16 tumor model

0 1. Implant 1x106 B16

cells in right flankC57BL/6

4

Day

7

23

2. Implant 5x104 B16

cells in left flank

3. PBS or 1x108 PFU

VV into right flank

Experiment Design

Right Flank Left Flank

Days post tumor cell implantation

D5 D7 D9 D11D0 D13 D15 D17 D19 D21 D23

Days post tumor cell implantation

D3 D5 D7 D9D0 D11 D13 D15 D17 D19

Tu

mo

r V

olu

me

(mm

3)

Tu

mo

r V

olu

me

(mm

3)

2500

2000

500

1500

1000

0

2500

2000

500

1500

1000

0

Results

FAP-TEA-VV inhibited growth of primary and secondary tumors

PBS

15

FAP-TEA-VV: Inhibition of Tumor Metastases in vivo

FAP-TEA-VV inhibits surface metastases and tumor growth in the B16 tumor model

0

50

100

150

200

PB

S

Eph

A2-

TEA

-VV

GM

CS

F-V

V

FAP

-TE

A-V

V

Mea

n #

of S

urfa

ce

Met

asta

ses

B

*

2x105

B16 i.v.

C57BL/6

1Day 0

PBS or 1x108 PFUs

VV i.v.

3 15

A

PBS

EphA2-

TEA-VV

GM-CSF

-VV

FAP-

TEA-VV

sacrifice and

harvest lungs

CExperiment Design

0

50

100

150

200

PB

S

Eph

A2-

TEA

-VV

GM

CS

F-V

V

FAP

-TE

A-V

V

Mea

n #

of S

urfa

ce

Met

asta

ses

B

*

2x105

B16 i.v.

C57BL/6

1Day 0

PBS or 1x108 PFUs

VV i.v.

3 15

A

PBS

EphA2-

TEA-VV

GM-CSF

-VV

FAP-

TEA-VV

sacrifice and

harvest lungs

C

0

1. Intravenous injection of

2x105 B16 cellsC57BL/6

1

Day

3

15

2. Inject PBS or 1x108

PFUs VV

3. Sacrifice and harvest lungsM

ean #

of

Surf

ace

Met

asta

ses

FAP-TEA-

VV

GM-CSF-

VV

EphA2-

VV

PBS

Results

Inhibition of Tumor Metastases

In the same tumor

model, FAP-TEA-VV

inhibited tumor

metastases compared

to virus alone

Murine Construct

16

FAP-TEA-VV: T cell infiltration in vivo

Murine Construct

anti-CD3 DAPI Merge

PBS

EphA2-

TEA-VV

GM-CSF

-VV

FAP-

TEA-VV

C

A B

NS

P < 0.05

P < 0.01

NS

P < 0.05

P < 0.01

%ofCD

8+tum

orinfiltratingcells

PBSEphA2- GM-CSFFAP-TEA-VV-VVTEA-VV

%ofCD

4+tum

orinfiltratingcells

PBSEphA2- GM-CSFFAP-TEA-VV-VVTEA-VV

anti-CD3 DAPI Merge

C

B16 subcutaneous tumor model

3 intratumor VV injections (D7, D10,

D13), Tissue harvest and assay (D15)

A&B) Infiltrating T cells by flow

cytometry

C) Infiltrating T cells by fluorescent

microscopy

17

FAP-TEA-VV: T cell activation in vivo

Murine Construct

B16 subcutaneous tumor model

3 intratumor VV injections (D7, D10,

D13) Tissue harvest and assay (D15)

D) Intracellular staining of tumor

infiltrating T cells

E) ELISPOT IFNγ of splenic CD8+ or

CD4+ T cells against dendritic cells

presenting FAP or TRP2 (a B16

tumor antigen)E

0

50

100

150

200

250

300

Me

diu

m

Ep

hA

2-T

EA

-VV

GM

-CS

F-V

V

FA

P-T

EA

-VVS

FC

pe

r 5

x1

04

CD

8+

T c

ells

Medium

DC-Lv-contr.

DC-FAP-Lv

DC-TRP2-Lv

0

40

80

120

160

Me

diu

m

Ep

hA

2-T

EA

-VV

GM

-CS

F-V

V

FA

P-T

EA

-VVS

FC

pe

r 5

x1

04

CD

4+

T c

ells

Medium

DC-Lv-contr.

DC-FAP-Lv

DC-TRP2-Lv

P <0.05P <0.05

PBS EphA2-TEA-VV GM-CSF-VV FAP-TEA-VV

CD4-PerCP

IL-2

CD8-PE

IFN

-γ

FSC

SS

CD

0%

0%

1.14% 6.36% 12.3%

7.35% 10.9% 9.58%

D

18

FAP-TEA-VV: Correlation of Stromal Destruction & Virus Spread

Anti-FAP DAPI Merge

PBS

EphA2-TEA-VV

GM-CSF-VV

FAP-TEA-VV

A

0

5

10

15

20

PB

S

Ep

hA

2-T

EA

-VV

GM

-CS

F-V

V

FA

P-T

EA

-VV

% F

AP

+ c

ells

B **

0

5

10

15

20

PB

S

Ep

hA

2-T

EA

-VV

GM

-CS

F-V

V

FA

P-T

EA

-VV

VV

tite

r(x

10

8P

FU

s/g

tissue)

C

0

5

10

15

20

0 4 8 12 16

%F

AP

+ c

ells

VV titer (x108 PFUs/g tissue)

PBS

EphA2-TEA-VV

GM-CSF-VV

FAP-TEA-VV

D

**

Murine Construct

B16 subcutaneous tumor model

3 intratumor VV injections (D7,

D10, D13). Tissue harvest and

assay (D15)

A) Fluorescent microscopy of

FAP+ cells

B) % of FAP+ cells in tumor tissue

C) VV titer per gram of tumor

tissue

D) Relationship between FAP+

cells and VV titer

19

FAP-TEA: mTEA-VVs have no systemic anti FAP activity

Murine Construct

B16 s.c. & i.v. tumor model

Serum was collected from mice treated with

PBS or GFP-VV or EphA2- TEA-VV or

mFAP-TEA-VV for both i.v. and s.c. models

(n = 5). Supernatant was collected from MC-

38 cells infected with GFP-VV or EphA2-

TEA-VV or mFAP-TEA-VV at MOI 5 for 24

h. Supernatant served as positive control

Medium served as negative control. GL-261-

FAP-GFP cells were co-cultured with Con A-

activated mouse splenocytes in presence of

supernatant or serum for 24 h. The killing

activity was analyzed by flow cytometry

GFP Expressing

FAP+ Target Cells

20

FAP-TEA: TEA-VV Does Not Affect Bone Marrow Cellularity or Mouse Weight- metastatic B16 model

Murine Construct

mFAP-TEA-VVs do not affect the total cell number of mice bone marrow and mice weight in systematic metastasis B16 model. 2 × 105 B16 cells

were i.v. injected through tail vein of C57/BL6 mice (n = 5). The mice were injected 1 × 108 VV through tail vein on day 1 and 3. (a) On day 15,

bone marrow was collected for counting of total bone marrow cell numbers. (b) Mice body weights were monitored at the different time points

A B

• Complete Phase I/II trials

21

R&D BCM/CAGT

2013 2014 2015 2016 2017 2018-2019 2020+

Provisional Patent Angel Round

Joined JNJ JLABS Incubator

3 Additional Patents Filed IND Application

Phase I StudyIND Enabling Studies

• Licensed TEA-VV patent from BCM & filed 3 additional patents

• IKT is working with Novella to file IND application in 2018

• IKT’s lead product (clinical grade VV) will be produced in Q4 2017

2015-2017

• Initiate phase I studies (IT/IV injection) in USA in Q2 2018

2018-2019

2019+

Rationale for FAP-targeted VV therapy

22

Limitations HSV-GMCSFVV-GMCSF

(Wyeth Strain)

FAP-TEA-VV

(WR Strain)

Tumor Stroma ▼Limits viral spread/oncolysis

▼Limits viral spread/oncolysis

▲Targeted by FAP-TE, enhancing viral spread/oncolysis

Endogenous anti-Tumor T cell

Required▲Enhanced by GMCSF

Required▲Enhanced by GMCSF

▲Not required▲TE redirects any T cell to target

Anti-viral Immunity ▼Enhanced by GMCSF

▼Enhanced by GMCSF

▲Anti-viral T-cells may be redirected to antigen target

Immune Suppressive MDSCs

▼Enhanced by GMCSF

▼Enhanced by GMCSF

▲Targeted by FAP-TE

Infectious Spread Receptor mediated ▲Efficient cell-cell▼Limited by CAF

▲Efficient cell-cell▲Enhancing spread through CAF

Viral Replication ▼Requires nuclear translocation

▲In cytoplasm within 2 hours

▲In cytoplasm within 2 hours

Shautong Song, MD/PhD

CEO, Icell Kealex Theapeutics

JLABS@TMC

2450 Holcomb Blvd J206, Houston, TX 77021

www.icellkealex.com

shautong.song@icellkealex.com

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Thank You!!!