Lawrence G. Lum, MD, DSc Director of Immunotherapy

Scientific Director of BMT

Co-Leader Tumor Microenvironment

Professor of Oncology, Medicine,

Pediatrics, and Immunology & Microbiology,

Karmanos Cancer Institute and

Wayne State University, Detroit, MI

Workshop , September 22, 2015

BiAbs, BITES, and BATs?

BiAb targeting for liquid tumors?

BiAb targeting for solid tumors?

BiAbs loaded onto effector cells (T cells, NK, monocytes, or PMNs) for:

1. Leukemias

2. Solid Tumors

Targeting Solid and Liquid Tumors

1. Can BiAb target stem cells or other tissue repair

cells to injured tissue?

2. Can BiAbs be used to armed “stem-like” or

other tissue repairs cells to injured tissues

3. What targets would you select?

Tissue Regeneration

1. Can BiAb target viral diseases?

2. Can BiAbs be used to armed immune effectors

to clear viral disease?

3. What targets and effectors pairs would you

select?

Targeting Viral Diseases

Overview

1. Example of targeting solid tumors (catuzumab

approved in Europe for IP injection in ovarian

cancer) and why not more?

2. Example of targeting liquid tumor (blinatumumab

approved for ALL) and why not more?

3. Arming T cells to target breast, prostate, lung,

ovarian, neuroblastoma, osteosarcoma and other

GD2.

4. Arming T cells with anti-CMV to treat CMV

infections

5. Arming stem cells to repair myocardial infarcts

Challenges

1. Picking the right effector and targets

2. Picking the optimal or right route of administration

3. Picking the right clinical indicating to develop the drug

4. Developing the appropriate phase I/II trial that will lead

to registration

5. Avoiding dose limiting toxicities (avoiding off-target

effects)

6. Convincing VC funding of the appropriate timing and

path for entering the clinic.

7. Understanding the regulatory environment

Targeting of Nil Expression of Her2 on Sum 1315 Cells

+

Anti-CD3 Anti-CD20

=

Anti-CD3 x Anti-CD20

Arming of T

Cells

Tumor

Kill T Cell

Targeted Killing by T cells with BiAbs

14 d T cell

Expansion

Anti-CD3 Chemical Heteroconjugation

Cytokine/Chemokine

Secretion

Killing of Breast Cancer Cellsby Armed T Cells

E:T

0 5 10 15 20 25 30

% S

pe

cif

ic K

illi

ng

0

20

40

60

80

100

No Ab

0.5 ng Her2Bi

5 ng Her2Bi

50 ng HerBi

Killing of MCF-7 Cells by Armed T Cells

Sen et al., J Hematol. & Stem Cell Res 10:247, 2001

Production of Armed T cells

OKT3 (20 ng/ml) + 100 IU/ml of IL-2

PBMC from Pheresis

ATC are split every other day

Harvest, Arm with BiAb and Cryopreserve after 10-14d

Testing for cytotoxicity and cytokine production

Quality Control (Bacteria, fungal,

and Mycoplasma stain) 7 days

Treatment Schema for Stage IV Breast

GM-CSF 250 ug/m2/dose

IL-2 300,000 IU/m2/day

Wk1 Wk2 Wk3 Wk4 Wk8

3 Wks

Dose escalation:

5, 10, 20, 40 in standard

3+3 design

Table 1: Patient Characteristics

No. %

Age

< 50

≥ 50

14

9

60.9

39.1

Cancer Stage

Stage IV

23 100

Performance Status

(ECOG)

0

1

2

18

5

0

78.3

21.7

0

ER/PR Status

Positive

Negative

Unknown

14

8

1

60.9

34.8

4.3

HER2/neu Status

0

1+

2+

3+

Unknown

10

2

2

8

1

43.5

8.7

8.7

34.8

4.3

Prior Treatment w/

Herceptin

Yes

No

8

15

26.0

74.0

Stage IV Breast Cancer Patients

Stage IV BrCa Phase I Toxicities

Toxicity Grade Grade

1

Grade

2

Grade

3

Grade

4

Total

Episodes

% of

Total

Chills 0 4 36 0 40 51

Headache 0 3 14 0 17 22

N/V 8 1 2 0 11 14

Fever 3 1 0 0 4 5

Hypotension 1 3 0 0 4 5

Hypertension 0 0 0 1 1 1.3

SOB 0 1 0 0 1 1.3

Total 12 14 52 1 77

1 patient died of CHF related to digoxin toxicity after IT

was completed.1 patient developed a subdural hematoma

that was evacuated without complications

Clinical Responses to Her2Bi-Armed ATC Infusions

Clinical Responses to Her2Bi-armed ATC by Dose Levela

Response

(%)

All

Pts #

All Pts

%

Dose

Level 1

Dose Level

2

Dose

Level 3

Dose

Level 4

PR 1 4.3 0 1(100)c 0 0

SD 12 52.2 4(33.3) 2(16.7) 6(50) 0

PD 8 34.8 4(50) 3(37.5) 1(12.5) 0

NEb 2 8.7 1(50) 0 1(50) 0

a At one month follow-up after the last infusion and 14.5 weeks after last Tx. bDid not complete

infusion schedule or died before 1 month follow-up. cPt received only 80 billion cells due to slow

expansion. Evaluation 15 weeks after last chemotherapy/hormone therapy

Her2/neu negative Pt: PR 7 months post IT

Phase I: Metastatic Breast

Stage IV BrCa (n = 9)

0 1 2 3 4 5 6 7 80.1

1

10

100

1000

10000

100000

Infusion #

IL-1

2 (

pg

/ml)

Immune Responses to Her2Bi-Armed ATC Infusions and Overall Survival

CD20Bi

MIP-1

GM-CSF

Tumor

Monocyte

TAA

DC

DC IL-2

IL-4

IL-12

IFN

T cells

Effector Memory T cell Central Memory

IFN

TNF

TAA

HLA Class I/II

Endogenous T cells

Armed T Cell

In Situ Vaccination

BATs Overcome Tumor Induced Suppression

Immunosuppression

M2

Myeloid derived

suppressor cells

(MDSC)

T regulatory cells

(Treg)

CD3 xTAABi

Regression

IFN

TNF

TAA

Armed

T Cell BAT-

induced

Th1

cytokines

MIP-1

GM-CSF

Monocyte

IL-12

M1

M1 TAM : • T and NK

cytotoxicity

• chemosensitivity

• regression

M2 tumor-associated

macrophages (TAM):

• immunosuppression

• invasion/metastasis

• vascular remodeling

• chemoresistance

Progression

T & NK effectors

Tumor

IL-12

Wk1

AT

C

Wk2

AT

C

Wk3

AT

C

Wk4

AT

C

PB

SC

T

Preparative

Regimen

Phase I Infusions of ATC Armed With CD20Bi after

AutoPBSCT for NHL

Dose Level 1: 10 billion/infusion Total 40 billion

Dose Level 2: 15 billion/infusion Total 60 billion

Dose Level 3: 20 billion/infusion Total 80 billion

Le

uk

op

he

res

is

G-CSF

Priming

5 days

AT

C E

xp

an

sio

n

Cry

op

res

erv

ati

on

14 days

Start as early a day +4 after PBSCT

Le

uk

op

he

res

is

Funded by Leukemia and Lymphoma Society

0 10 20 30 40 50 60 700

10

20

30

40

50

60

70

80

90

100

110

All Pts

Rem

Non-Rem

Months post SCT

Perc

ent

surv

ival

Targeted T cells for NHL: Toxicity, Engraftment, CTL Activity, and Overall Survival

BATs Clear Neuroblastoma from Bone Marrow.

Before

After

0

5

10

15

20

25

30

35

40

45

50

25:1 12.5:1 6.25:1 3.125:1E:T Ratio

% C

yto

tox

icit

y

UA 33 Arm 0.01 Arm 0.1 Arm 1.0 Arm 5 Arm 10 Arm 100 IVIG Bi 50

Ramesh ASH Abstr 1157 (2009)

CMVBi Dose Titration (MOI of 0.1)

Specific cytotoxicity of CMVBi Armed ATC

0

20

40

60

80

100

120

25:1 12.5:1 6.25:1 3.125:1

% C

yto

tox

icit

y

CMVBiab

Cytogam

CMVBiab-1-ADCC

Cytogam-1-ADCC

CMVBiab-2-ADCC

Cytogam-2-ADCC

CMVBiab-UA-ATC 1

CMVBiab-ARM-ATC1

CMVBiab-UA-ATC2

CMVBiab-ARM-ATC2

E:T Ratio

Binding CMVBi-armed ATC

BiAb Production

S N + H 2 Cl - NH 2

N

O

O

CH 2 C

O

O N

O

O

SO 2 Na

N

O

O

CH 2 C

O

H N

H 2 N

N

O

O

CH 2 C

O

H N

N

N + H 2 Cl -

H

S

N

N + H 2 Cl -

H

SH

Traut’s reagent

Anti-CD45 or

Anti-CD3 Sulpho - SMCC

Anti-CD45 x anti-MLC

1 2

3

S N + H 2 Cl -

S N + H 2 Cl - NH 2

N

O

O

CH 2 C

O

O N

O

O

SO 2 Na N

O

O

CH 2 C

O

O N

O

O

SO 2 Na

N

O

O

CH 2 C

O

H N N

O

O

CH 2 C

O

H N

H 2 N H 2 N

N

O

O

CH 2 C

O

H N

N

N + H 2 Cl -

H

S N

O

O

CH 2 C

O

H N N

O

O

CH 2 C

O

H N

N

N + H 2 Cl -

H

S N

N + H 2 Cl -

H

S

N

N + H 2 Cl -

H

SH N

N + H 2 Cl -

H

SH

Traut’s reagent

Anti-MLC or

Anti-ICAM-1

1 2

3

+

Anti-CD45 Anti-MLC

=

Anti-CD45 x Anti-MLC

Armed Stem Cell Stem

Cell

Targeting Stem Cells with BiAbs

Injured Target

Tissue

A Xenogeneic Rat Model

TROPONIN TROPONIN Class I TROPONIN/Class I

Armed

Unarmed

4x 20x 20x 20x

20x 20x 20x 4x

Co-localized Staining of Cardiac and Human Class I in MIs of Rats IV Treated with

CD45 x -MLC-armed CD34

5 weeks Post-Internal Jugular Vein Injection

(2 x 106 G-CSF primed CD34+ Cells)

0 10 20 30 400.00

0.25

0.50

0.75 unarmed

armed

Time after occlusion (days)

fra

ctio

na

ls

ho

rte

nin

g

0 10 20 30 400.00

0.25

0.50

0.75

unarmed

armed

Time after occlusion (days)

sy

sto

lic

dim

en

sio

n

0 10 20 30 400.10

0.15

0.20

0.25 unarmed

armed

Time after occlusion (days)

dia

sto

lic

po

ste

rio

rw

all th

ick

ne

ss

0 10 20 30 40

0.1

0.2

0.3 unarmed

armed

Time after occlusion (days)

sy

sto

lic

an

teri

or

wa

ll t

hic

kn

es

s

Echo Results for Nude Rats Receiving CD34+ cells Armed with anti-CD45 x anti-MLC

9/19/04

Those who made it Happen!

Co-Is: A Thakur, A Deol, L Ayash, M Abidi, Z Al-Kadhimi, V Ratanatharathorn, J Uberti, J Zonder, A Weise

Lab Staff: E Tomaszewski, D Schalk, V. Kondadasula, H Yano , C Pray

Support Staff: P Steele, K Meyers , K Fields, M Dufresne, BMT and IV infusion nurses

This work was supported by grants: R01 CA 092344, R01 CA 140314, Leukemia & Lymphoma Society Grants Translational Awards, Michigan Life Science Grant, DOD, Young Foundation, and The Ruth F. Rattner and Ann F. & Norman D. Katz Foundation, KCI startup funding