Pengfei Zhou(MD, Ph.D)

Wuhan YZY Biopharma

Feb, 28th, 2016

A B

Bispecific antibody development

for tumor immunotherapy

Cancer Miracles 3 years cancer free! (Emily Whitehead)

Emily on stage at the Leukemia Ball with Dr. Carl June who

developed the T-cell therapy that saved her life

CD19 CART

2

Cancer Miracles Blinatumomab by Amgen

The price tag for this therapy is US$ 178,000 for two month treatment

3

Multiple molecular pathways for cancer(I)

Monotherapy vs. Multiple-targeting

Hanahan D, Cell, 2000, 100:57-70

4

Misunderstandings in Oncological Clinics (II) Ignored Metastasis Pathway

Primary

Lesion

Metastasis

Lesion

5

The Challenges of Precision Oncological

Medicine 1. Over 800 anti-cancer drugs are

in clinical developing which

target “the Hallmark of Cancer”

2. Target population has to be

identified for specific drug.

3. Biopsy sample is the golden

standard and major source of all

the pathology sample. But it’s

hard to get, not even mention to

follow up.

4. Circulating Tumor Cells (CTC)

had been regarded as one of the

reliable sample sources for the

personalized diagnosis and

therapy. Hanahan and Weinberg; Cell 2011; 144:1-29

The Hallmark of Cancer

6

Cancer Screening

• Cancer Biomarkers

• Immune biomarkers

Cancer Relapse

• CTC-Biopsy

Cancer Treatment

• Bispecific antibody

• immunotherapy

YZYBIO: R&D Strategy

Companion

Diagnostic Test Kits

Target Mutations:

KRAS/EGFR/BRAF

KIT/TP53/FLT3/JAK2

CYP2C9/CYP2C19/CY

P2D6/VKORC1

…….

7

Selected bispecific antibodies in clinical trials (2015.4.28)

Developers Lead molecule Targets Technology Indications Stage

Trion Pharma, Neovii

Biotecha(Munich) Removab

Epithelial cell

adhesion molecule

(EpCam) × CD3

Triomab quadroma technology

comprising hybrid mouse IgG2a ×

rat IgG2b antibody with intact

immune effector functions

Malignant ascites EU approval

Apr. 23, 2009

Amgen (Thousand

Oaks, California) Blincyto CD19 × CD3

Bispecific T-cell engager (BiTE)

antibodies comprising different

minimal antigen-binding domains

from two single-chain Fvs (scFvs)

arranged in tandem on a polypeptide

chain

Philadelphia

chromosome–

negative acute

lymphoblastic

leukemia

FDA approval

Dec. 3, 2014

Amgen AMG-110 EpCam × CD3 BiTE antibody Solid tumors Phase 1

AbbVie ABT-122 TNF-a x IL-17

DVD-Ig comprising tetravalent

bispecific antibody with two

additional variable domains,

attached by linkers to the N-termini

of the VH and VL domains of a

conventional monoclonal antibody

Rheumatoid

arthritis Phase 2

AbbVie ABT-981 IL-1a × IL-1b DVD-Ig Osteoarthritis Phase 2

Affimed Therapeutics AFM13 CD30 × CD16A

Tetravalent bispecific tandem

diabody (TandAb) comprising four

Fv domains joined by peptide linkers

Hodgkin lymphoma Phase 2

Nature Biotechnology, 33,219–221(2015)

8

9

Developers Lead molecule Targets Technology Indications Stage

Merrimack

Pharmaceuticals

(Cambridge,

Massachusetts)

MM-111

Human epidermal

growth factor receptor

2 (HER2) × HER3

Bispecific antibody fusion protein

comprising two scFv domains

linked by modified human serum

albumin

Gastric cancer Phase 2

Sanofi (Paris) SAR156597 IL-4 × IL-13 Tetravalent bispecific tandem

immunoglobulin (TBTI)

Idiopathic pulmonary

fibrosis Phase 2

Roche (Basel) RG7221

(RO5520985)

Angiopoietin 2 (Ang-

2) × vascular

endothelial growth

factor a (VEGF-A)

Bispecific IgG1 Crossmab

antibody based on knob-into-hole

mutations and Fab domain

exchange

Colorectal cancer Phase 2

Roche, Chugai

(Tokyo) RG6013 (ACE910) Factor IXa × factor X

Asymmetric bispecific IgG4

antibody Hemophilia A Phase 2

Genentech (S. San

Francisco,

California)

RG7597

(MEDH7945A)

Epidermal growth

factor receptor

(EGFR) × HER3

Two-in-one IgG1 antibody

KRAS wild-type metastatic

colorectal cancer;

recurrent/metastatic head

+ neck cancer

Phase 2

Ablynx (Ghent,

Belgium), Merck

Serono (Darmstadt,

Germany)

ALX-0761 IL-17A × IL-17F Bispecific nanobody with

albumin-binding domain Autoimmune disease Phase 2

Merus (Utrecht, the

Netherlands) MCLA-128 HER2 × HER3

Biclonics full-length bispecific

antibody Solid tumors Phase 1/2

AstraZeneca

(London), Amgen

MEDI-565 (AMG-

211)

Carincoembryonic

antigen × CD3 BiTE antibody

Gastrointestinal

adenocarcinoma Phase 1

Macrogenics, Servier

(Suresnes, France) MGD006 CD123 × CD3

Dual-affinity retargeting (DART)

bispecific antibody based on two

covalently linked Fv polypeptides

Acute myeloid leukemia Phase 1

Regeneron

(Tarrytown, New

York)

REGN1979 CD20 × CD3 Bispecific antibody Advanced malignancies Phase 1

Continued 9

Catumaxomab (Removab®) Blinatumomab (Blincyto®)

Time EMA, 2009/4/20 FDA, 2014/12/3;

EMA, 2015/11/26

Target EpCAM×CD3 CD19×CD3

Indication Malignant ascites ALL

Stucture

MOA T cell activation, ADCC, CDC,

phagocytosis T cell activation

Cost • 12,000 euros（7 days）

• 4 times：10、20、50、150µg/dose

•178,000 US$（28 days）

•28 days：9µg/day×7，28µg/day×21

Limitation •Mouse/rat origin: Hypersensitivity, PK, PD；

•Low production capacity;

•Side effects；

•Short half time (lack of Fc region);

• Continuous intravenous infusion;

•Side effect;

Characteristics of Approved BsAbs 10

Name Company Structure Characteristics Technology

challenges Products Deal

BiTE MicroMET Small size;

Good permeability;

No mismatch.

Short halftime;

Aggregation;

Difficult purify；low affinity；

Blinitumomab

（CD19xCD3）；MT110

（EpCAMx CD3）;

MT111（CEAx

D3）；

MT112

（PSMAxCD3）

Bayer, Sanofi,

Boehringer ,

AMGEN;

$1.9B

Triomab Trion

/Fresenius AG

IgG like structure;

good PK;

stability；

High affinity;

High

Immunogenicity；

Low expression;

Catumaxomab

（EpCAMxCD3）；

Ertumaxomab

（HER2xCD3）；

FBT-A05 (CD20xCD3)

N/A

Duobody Genmab IgG like structure;

good PK; Difficult product; N/A

Novartis，$175M；

Johnson &

Johnson‘s，

$3.6B

DVD-Ig AbbVie

IgG like structure;

Good PK;

Can expressing in

mammalian cell

Large size;

ABT-122

ABT-981 N/A

DutaMab

Dutalys IgG like structure;

No mismatch.

Difficult

development;

Influenced affinity

N/A Roche，$489M

Bispecific Antibody Platform (1)

11

Name Company Structure Characteristics Technology

challenges Products Deal

DART MacroGenics

Small size;

No linker between VH and VL;

Disulfide bond;

High stability.

Short half-life;

Probability of

mismatch;

Influenced affinity;

No ADCC.

MGD010(CD32BX

CD79B);

Boehringer

Ingelheim, $2.1B;

Gilead Science ,

$1.1B

Takeda,$500

million;

CrossMab Roche

Like a full-length antibody;

High affinity;

Binding to Fc receptors.

Difficult to remove

homodimer;

N/A N/A

Fcab F-star

Excellent binding;

Favourable pharmacokinetics;

mobilise immune effector

functions;

Protein stability;

Simplicity of manufacturing.

Not suitable for

every target;

Limited Affinity;

Influenced ADCC

effect.

6 drugs

Boehringer

Ingelheim , $1.7B

Merck Seronol ,

$708M

SEED Merck

serono

Potency like a full-length

antibody;

High affinity;

Hight stability;

Good half-life.

Influence ADCC

effect;

Difficult to remove

homodimer;

N/A N/A

Azymetric Zymeworks

tailored effector function;

ptimized serum half-life;

not rely upon chemical linkers

or chimeric constructs

Difficult to develop N/A MSD， $178M

Bispecific Antibody Platform (2)

Antigen

binding sites

S-S

12

YBODY Structural Property

13

Characteristics of YBODY

1. Whole IgG: good PK and PD, facilitate purification；

2. Knock-in-hole and salt bridge strategy: high efficiency of

heterodimerization production；

3. Unique MOA, potency improved over 1000 times；

4. Tumor immunotherapy, reduce the recurrence；

5. Low dosage, reduced side effects and cost

6. Platform technology;

Anti-CD3

TSA/TAA

human IgG1 Fc

region

human IgG1 Fc

region

14

GFP +肿瘤细胞

被淋巴细胞包围

保密文件

MW=130Kd

YBODY Unique Heterodimerization Strategy

CH

3

CH

2 C

H2

S S

S S

CH

3 C

H3

C

H2

CH

2

+ -

S S S S

CH

3

CH

3

CH

2 C

H2

+ -

S S

S S

CH

3

Knob-into-hole Salt-bridge

Knob-into-hole + Salt-bridge

High-efficiency of heterodimerization

15

保密文件

M802 (anti-HER2×CD3 BsAb)

Indication：Breast cancer, gastric cancer

16

Expression of HER2 in Different Tumors

Tumor Her2 +++ Her2 ++ Her2 +/-

Breast tumor 20% 30% 50%

Gastric tumor 47% [1] 30% [1] 23% [1]

Ovarian tumor 1.8% [4] 15% [4]

Non-small cell lung tumor 16-21% [4]

Colorectal tumor 66.7% (stage B), 58.6% (stage C),

28.6% (stage D) [3]

Bladder tumor 26% [2]

Pancreatic tumor 44% [2]

Wilms' tumor 51% [2]

[1]. http://www.cancer.gov/cancertopics/druginfo/fda-tratuzumab#anchor-Breast

[2]. S. Ménard, et al., Ann Oncol (2001)

[3]. K. Ghaffarzadegan, et al., IJBMS, Vol. 9, No. 1, Spring 2006

[4]. Robert O. Dillman. Principles of Cancer Biotherapy (5th ED, 2009)

17

保密文件

2013/2014 Global Sales of HER2-

Targeted Antibodies

Antibody Drug Appro

ved

Indication 2013/2014

sales

（$ milllion） FDA[1] EMA[2]

Trastuzumab

(Herceptin) Roche 1998

1.Adjuvant breast cancer；2.Metastatic breast cancer ；3. Metastatic Gastric cancer

1. Breast Neoplasms;

2. Gastric Neoplasms 6557/6490

Pertuzumab

(Perjeta) Roche 2012

1. Metastatic breast cancer；2. Neoadjuvant treatment of

breast cancer

1.Breast Neoplasms 352/940

Ado-trastuzumab

emtansine (Kadcyla) Roche 2013 1. Metastatic breast cancer 1.Breast Neoplasms 252/535

18

保密文件

[1] http://www.fda.gov/

[2] http://www.ema.europa.eu/ema/

Significant Unmet Need for New HER2 Drugs

• High proportions of HER2(+) BC patients do not respond:

• Trastuzumab: 20-50% non-responders in adjuvant setting

• Trastuzumab : 70% non-responders as monotherapy

• T-DM1 ~35% non-responders

• Most responders eventually progress, presumably some due to

acquired resistance.

• Trastuzumab dosen’t have a good performance in Patients with

low HER2 expression

19

Confidential

M802 cross-reacts with human CD3 and Her2 positive

tissues

Human Her2 positive breast tissue Human Thymus

20

保密文件

Compared to monoclonal antibodies against Her2 or CD3, M802 BsAb mediated

strong cytotoxicity to Her2 lowly expresssd tumor cells such as MCF-7 and MDA-

MB231, indicating broader clinical applications.

MCF-7 (Breast cancer) /CIK (Cytokine-induced killer)24h （E:T=5:1）

10-1 100 101 102 103 104 105

0

20

40

60

80

100M802

Herceptin

L2K (Anti-CD3)

Antibody conc.(pg/ml)

Cyt

oto

xic

ity

(%)

MDA-MB-231(Breast cancer) /CIK (Cytokine-induced killer)24h （E:T=5:1）

10-1 100 101 102 103 104 105

0

20

40

60

80

100M802

Herceptin

L2K (Anti-CD3)

Antibody conc.(pg/ml)

Cyt

oto

xic

ity

(%)

21

Cytotoxicity of M802 to Her2 lowly expressed cell

strain (1)

保密文件

cell line EC50(pg/ml)

SK-BR-3 3938

BT474 9443

MDA-MB-453 9067

MCF7 4512

MDA-MB-231 40671

Cytotoxicity of M802 to Her2 lowly expressed cell

strain (2)

保密文件

22

PK of M802 preclinical study

Animal：Babl/c

Route：IV, 4mg/kg；

Drug Name Half-life ((h)

Blinatumomab (Amgen) 1.25 (human)

M802 64.32 (mouse)

0 100 200 300 400 500100

1000

10000

100000M802 4mg/kg

Time (hours)

Seru

m d

rug

co

nc.(

ng

/ml)

Animal：6 cynomolgus monkey each panel

给药方式 ：IV, 0.3、1、3mg/kg；

Drug Name Half-life ((h)

Trastuzumab 144 (Monkey)

M802 92 (monkey)

23

保密文件

0 100 200 300 400 500 600 700

100

101

102

103

104

105

106

Co

ncen

trati

on

(n

g/m

L)

Time (h)

M802 (0.3 mg/kg)

M802 (1 mg/kg)

M802 (3 mg/kg)

Tumor growth inhibition of M802 to subcutaneously transplantated

NCI-N87

Animal Model: The NOD-SCID mouse was subcutaneouly transplanted with NCI-

N87 tumor cells and human CIK cells, followed by IV injection of M802.

24

保密文件

0 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 51 54 57 60

0

250

500

750

1000

1250

1500

生理盐水

Herceptin 5mg/kg

M802 5mg/kg

M802 2mg/kg

M802 1mg/kg

M802 0.5mg/kg

0 2 4

天

肿瘤

体积

/ mm

3

Animal Model: The constructed Her2 positive CT-26 in house was intravenously injected to

Balb/c, followed by IV injection of M806 (anti-human Her2×anti-mouse CD3)

Survival Curve of M806 to Babl/c intravenouly injected

with CT26-Her2

25

0 10 20 30 40 500

20

40

60

80

100

生理盐水 0/10

Herceptin (5mg/kg) 0/10

M806（0.5mg/kg) 4/10

M806（0.25mg/kg) 9/10

M806（0.05mg/kg) 10/10

M806（0.01mg/kg) 7/10

M806（0.002mg/kg) 7 /10

天

生存

率%

Preclinical Evaluation:

• No Adverse Effects were observed after ICR mice single IV injection with

M802 at 137.5 mg/kg

• 30 cynomolgus monkeys were divided into 3 panels, twice I.V. drip at 1, 3,

10 mg/kg each panel every week. After 4 weeks administration, except one

monkey produced HAMA(1/30), no other adverse effects were observed.

26

保密文件

27

保密文件

Optimization, In

Vitro/Vivo

Assay development

QC validation

Pharmacodynamics

Toxicity

I

N

D

CLD（CHO-S), Cell bank,

Process development, formulation

Pilot production

Quality Control

2013 2013 2014 2014 2014 2014 2015 2015 2015 2015

3rd 4th 1st 2nd 3rd 4th 1st 2nd 3rd 4th

Her2/CD3

IND Timeline for M802 BsAb

IND： Dec. 1st, 2015

受理号：CXSL1500125鄂

28

保密文件

M701(anti-EpCAM×CD3 BsAb)

Indication：Malignant ascites, metastatic ovarian cancer,

colorectal cancer

Cancers can cause malignant ascites

EpCAM: Epithelial Cell Adhesion Molecule

Tumor Positive rate

Ovarian tumor 88-91%

Gastric tumor 98%

Colorectal tumor 99%

Pancreatic cancer 96%

Breast tumor 90%

Endometrial 91-96%

Lung tumor 87%

Prostatic tumor 98%

The positive rate of EpCAM

expression in different tumors

Broad Therapeutic Applications for EpCAM-Targeted Drugs

29

Confidential

Model: CT26 was transfected to express human EpCAM, which was called CT26-EpCAM. The CT26-EpCAM was injected i.p.

into Balb/c mice to form syngeneic model. With CT26-EpCAM as the tumor cells, M706 was given by i.p. to mediate the mouse

immune cells to kill tumor cells. M706 was a mimetic bsAb which binds to EpCAM antigen in one side and mouse CD3 in the other

side.

Syngeneic Tumor Model

M706

PBS

27 days after administration

Mice： Balb/c

Group： 5 mice per group

Injected cells： CT26-EpCAM

Administration：i.p.

0 10 20 30 40 50 600

25

50

75

100

125PBS

Anti-EpCAM 0.125mg/kg

M706 0.125mg/kg

M706 0.0625mg/kg

Days after tumer challenge

Perc

en

t su

rviv

al

In this model, M706 (the mouse surrogate ) significantly prolonged

mouse survival in a dose-dependent manner.

30

Confidential

31

保密文件

2013 2013 2014 2014 2014 2014 2015 2015 2015 2015 2016

3rd 4th 1st 2nd 3rd 4th 1st 2nd 3rd 4th 1-2th

EpCAM/CD3

IND timeline for M701 BsAb

Optimization, In

Vitro/Vivo

Assay development

QC validation

Pharmacodynamics

Toxicity

IND

filing

CLD（CHO-S), Cell bank,

Process development, formulation

Pilot production

Quality Control

Target Indication Stage

Target

validation Engineering Potency CLD/PD Preclinical Clinical

Her2/CD3 Breast cancer,

gastric cancer

EpCAM/CD3 Malignant ascites

CD20/CD3 Lymphoma

CD19/CD3 Lymphoma

CD133/CD3 Colorectal cancer

EGFR/CD3 Colorectal cancer

EMP2/CD3 Ovarian cancer

PD-L1/CD3 Solid tumor

YZY YBODY Pipeline

32

保密文件

National Priority 12th five-year project (2015）

NSFC

National Priority 12th five-year project (2014）

YBODY armed CIK therapy (ACC)

Intellectual Property

BISPECIFIC ANTIBODY

International Application No. PCT/CN2012/084982

U.S. Patent No. US9079965B2

Taiwan Patent Application No. 102142238

China Patent Application No. 201280065551.5

Canada Patent Application No. 2892059

Europe 8 country Application No. 12888935.9

Priority Date: 21 November 2012

A CD3 ANTIGEN, A PREPARATION METHOD

AND APPLICATION THEREOF

一种CD3抗原及其制备方法和用途

Patent No. CN201310440453.1

A CD3 ANTIGEN, A PREPARATION METHOD

AND APPLICATION THEREOF

一种CD3抗原及其制备方法和用途

Patent No. CN201310399169.4

PREPARATION AND SELECTION OF CELLS

FOR PRODUCING BISPECIFIC ANTIBODIES

International Application No. PCT/CN2014/079303

Priority Date: 5 June 2014

BISPECIFIC ANTIBODY-MEDIATED CANCER

THERAPY WITH CYTOKINE-INDUCED KILLER

CELL

International Application No. PCT/CN2014/082590

Priority Date: 21 July 2014

一种鼠肿瘤模型的构建方法及其应用

China Patent Application No. 2015100299268

Priority Date: 21 January 2015

A SERIES OF BISPECIFIC ANTIBODY (Submitted)（a

total of 12 new antibodies including HER2XCD3,

EpCAMXCD3, CD19XCD3, CD20XCD3, CD133XCD3,

EGFRXCD3)

一系列双特异性抗体（共12个发明专利）

Priority Date: 21 January 2015

33

Confidential

Peer reviewed paper (CD133XCD3 YBODY®)

34

34

Journal of Immunotherapy 2015

友芝友生物制药与武汉协和医院肿瘤中心合作项目

国家自然科学基金资助项目

CD133XCD3 双靶向抗体

Summary

• Two drug with the price of one: Bispecific antibodies are the fastest

growing class of therapeutic agents.

• Excellent in vitro/in vivo pharmacology data (M802 and M701)

• Successful process development to meet the manufacturing

challenges

• YBODY provides an attractive alternative for Triomab/BiTE/DART

technologies

35

For Use with Sanofi Discussion Only

总结

• 双特异性抗体是研究进展最热的一类抗体.

• YBODY项目Her2xCD3和EpCAMxCD3具有优越的体内外肿瘤治疗生物学活性

• YBODY 中试生产工艺（培养、纯化、制剂等）成功开发，有利于大规模生产

• YBODY比Triomab/BITE/DART可能更具有技术优势

36

保密文件

周鹏飞

pfzhou@yzybio.com

37