Safety, Pharmacokinetics and Proof-of-Mechanism of an Oral Bruton’s Tyrosine Kinase Inhibitor HM71224 in Healthy Adult Volunteers Jimin Lee1, Jeewoong Son2, Hui Jung Sin1, Jongsoo Woo2, Salah Hadi3, Khee Hyun Suh4, Young-Mi Lee4, Sunyoung Jang4, Jin-A Jung1

1Clinical Research Team, Hanmi Pharm. Co., Ltd., 2Hanmi Pharm. Co., Ltd., Seoul, Korea, Republic Of, 3PRA HealthSciences, Zuidlaren, Netherlands, 4Hanmi Research Center, Seoul, Korea, Republic Of

Introduction

Conclusions • HM71224 demonstrated well-tolerated safety profile in healthy volunteers and

desirable PK and PD properties

• The data support the potential for HM71224 to be evaluated for treatment of

autoimmune diseases such as rheumatoid arthritis

THU0185

• Bruton’s Tyrosine Kinase (BTK) plays key roles in B-cell receptor (BCR) and Fc

receptor (FcR) signaling cascades and B cell development and activation1-8

• HM71224 is an orally available, irreversible and highly selective small molecule

inhibiting BTK protein

• HM71224 may provide therapeutic opportunities in autoimmune diseases

• Primary Objective: To evaluate the safety and tolerability and if possible to

determine the maximum tolerated dose of HM71224 after single and multiple

ascending dose administration in healthy subjects

• Secondary Objective:

a. To determine the pharmacokinetics(PK) of HM71224 and selected metabolites

(M1 and M2) following single and multiple oral dose administration of HM71224

b. To assess whether the PK of HM71224 is affected by food

c. To assess the occupancy by HM71224 after multiple oral administration of

HM71224 (Multiple Ascending Dose Part only)

Methods • Phase 1 study is consisted of 3 parts; a single ascending dose (SAD) part, a single

food effect (FE) part and a multiple ascending dose (MAD) part

• The SAD, FE and a part of MAD part results were revealed previously

• In the MAD part, once daily dosing and twice daily dosing were evaluated in placebo

controlled manner under fasted conditions

Results

• Total 90 subjects were included in the Phase 1 study

• 2 subjects were withdrawn due to adverse events in the MAD part; 88 subjects

completed the study per protocol

• No serious adverse events were reported in the Phase 1 study

• No clinically relevant changes in vital signs, ECGs and clinical laboratory tests

• All treatment emergent adverse events (TEAEs) were transient and resolved without

sequelae by follow-up

• Most common TEAEs were reported in Gastrointestinal system classified by system

organ

Reference 1. Bioessays. 2001 May; 23(5):436-46.

2. Nat Rev Immunol. 2002 Dec; 2(12):945-56.

3. Nat Rev Drug Discov. 2003 Jun; 2(6):473-88.

4. Immunol Rev. 2007 Aug; 218:45-64.

5. Immunol Rev. 2009 Mar; 228(1):58-73.

6. Nat Rev Rheumatol. 2009 Jun; 5(6):317-24.

7. Int Rev Immunol. 2012 Apr; 31(2):119-32.

8. Clin Immunol. 2013 Jul; 148(1):66-78.

Acknowledgement We would like to thank all of the participating patients and their families, as well as study

coordinators of the all study sites

This study was sponsored by Hanmi Pharmaceutical Co., Ltd. ClinicalTrial.gov identifier :

NCT01765478

• BTK occupancy was evaluated with BID dosing cohorts in MAD part

• More than 80% of BTK occupancy was maintained until 48 hours after the single

dose (Day 3, pre-dose)

• At steady state, more than 90% of BTK occupancy was achieved above 20mg BID

dosing

• BTK occupation lasted more than 7 days after completion of dosing (data not shown)

% BTK occupancy after 2 weeks

Dose (mg)

% B

TK

occu

pa

ncy a

t C

tro

ug

h

02

04

06

08

01

00

1 5 10 15 20 25 30 40

90

%

QD dose

BID dose

• A PK-PD model for HM71224 showed a steep dose response relationship with

BTK occupancy

• According to the PK-PD model above 10mg BID dosing or 20mg QD dosing shows

90% of BTK occupancy with HM71224

B-Cell Receptor Fcγ Receptor

B-cell Myeloid cell

- Proliferation - Cytokine production - Antibody Secretion

- Phagocytosis - Cytokine production

Bruton’s Tyrosine Kinase

Bruton’s Tyrosine Kinase

HM71224

Objectives

The 16th European League Against Rheumatism (EULAR) Annual European Congress, Rome, Italy; June 10-13, 2015

Sing Ascending Dose (n=18)

Food Effect (n=8)

Multiple Ascending Dose QD, 14-day (n=40)

Multiple Ascending Dose BID, 14-day (n=24)

10mg

20mg

40mg

80mg

140mg

60mg 10mg

20mg

40mg

80mg

200mg

120mg

40mg

5, 20mg

60mg

1. Subject Demography

Characteristics SAD Part

(n=18)

FE Part

(n=8)

MAD Part

QD

(n=40)

BID

(n=24)

Race (N, %)

Caucasian 17 (94.4%) 6 (12.5%) 32 (80.0%) 22 (91.7%)

Black - 1 (12.5%) 3 (7.5%) 1 (4.2%)

Asian, American

Indian or Alaska naïve 1 (5.6%) 1 (75.0%) 3 (7.5%) 1 (4.2%)

Others - - 2 (5.0%) -

Age, median (yr) 52.5 (±10.0) 28.5 (±19.8) 33.5 (±15.0) 33.0 (±12.6)

Height (cm) 182.4 (±7.8) 179.3 (±8.6) 179.6 (±9.7) 180.5 (±5.2)

Weight (kg) 81.7 (±11.5) 75.6 (±10.7) 79.3 (±11.8) 78.4 (±10.0)

BMI (kg/m2) 24.5 (±2.4) 23.5 (±3.0) 24.5 (±2.5) 24.1 (±2.9)

2. Adverse Events in Multiple Ascending Dose Part QD dosing BID dosing

Placebo

N=10

n (%)

[#AEs]

10 mg

N=6

n (%)

[#AEs]

20 mg

N=6

n (%)

[#AEs]

40 mg

N=6

n (%)

[#AEs]

80 mg

N=6

n (%)

[#AEs]

120 mg

N=6

n (%)

[#AEs]

Placebo

N=6

n (%)

[#AEs]

5 mg

N=3

n (%)

[#AEs]

20 mg

N=3

n (%)

[#AEs]

40 mg

N=6

n (%)

[#AEs]

60 mg

N=6

n (%)

[#AEs]

TEAEs

6

(60%)

[13]

3

(50%)

[3]

5

(83.3%)

[7]

3

(50%)

[3]

5

(83.3%)

[20]

5

(83.3%)

[18]

3

(50%)

[5]

2

(66.7%)

[3]

2

(66.7%)

[3]

4

(66.7%)

[7]

5

(83.3%)

[16]

Any ADRs - - - -

1

(16.7%)

[1]

4

(66.7%)

[12]

- - -

1

(16.7%)

[2]

-

Severity

Mild - - - - -

3

(50%)

[7]

- - -

1

(16.7%)

[1]

-

Moderate - - - - -

1

(16.7%)

[1]

- - -

1

(16.7%)

[1]

-

Severe - - - -

1

(16.7%)

[1]

3

(50%)

[4]

- - - - -

By System Organ

Class (SOC)

Skin/Subcutaneous

tissue disorders - - - - -

4

(66.7%)

[6]

- - -

1

(16.7%)

[1]

-

Nervous system

disorders - - - - -

1

(16.7%)

[2]

- - -

1

(16.7%)

[1]

-

Gastrointestinal

disorders - - - - -

1

(16.7%)

[1]

- - -

1

(16.7%)

[1]

-

General disorders/

Administration site

conditions

- - - - -

1

(16.7%)

[1]

- - - - -

Immune system

disorders - - - -

1

(16.7%)

[1]

- - - - - -

Musculoskeletal

/Connective tissue

disorders

- - - - -

1

(16.7%)

[1]

- - - - -

Vascular disorders - - - - -

1

(16.7%)

[1]

- - - - -

3. Pharmacokinetic Profiles

4. BTK Occupancy

• HM71224 showed increasing PK profiles as ascending dose levels in QD dosing

• HM71224 indicated slight accumulation after multiple dosing for 14 days

• Inter subject variability in exposure was relatively large

• Excretion of HM71224 and metabolites in urine was limited (data not shown)

* Only active data is included above.

Study Drug Administration

Day Morning

dose

Evening

dose

1 O X

2 X X

3-13 O O

14 O X

5. PK-PD Modeling

D1 P

red

ose

D1 4

hr

D1 1

2 hr

D3 P

red

ose

D3 2

hr

D6 P

red

ose

D1 7

D1 P

red

ose

D1 4

hr

D1 1

2 hr

D3 P

red

ose

D3 2

hr

D6 P

red

ose

D1 7

D1 P

red

ose

D1 4

hr

D1 1

2 hr

D3 P

red

ose

D3 2

hr

D6 P

red

ose

D1 7

D1 P

red

ose

D1 4

hr

D1 1

2 hr

D3 P

red

ose

D3 2

hr

D6 P

red

ose

D1 7

0

1 0

2 0

3 0

4 0

5 0

6 0

7 0

8 0

9 0

1 0 0

BT

K O

cc

up

an

cy

(%

)

0 4 8 1 2 1 6 2 0 2 4 2 8 3 2 3 6 4 0 4 4 4 8

0 .1

1

1 0

1 0 0

1 0 0 0

1 0 m g , M A D , D 1

1 0 m g , M A D , D 14

2 0 m g , M A D , D 1

2 0 m g , M A D , D 14

4 0 m g , M A D , D 1

4 0 m g , M A D , D 14

8 0 m g , M A D , D 1

8 0 m g , M A D , D 14

1 2 0 m g , M A D , D 1

1 2 0 m g , M A D , D 14

T im e (h r)

Pla

sm

a c

on

ce

ntr

ati

on

(n

g/m

L)

0

2 0 0

4 0 0

6 0 0

8 0 0

1 ,0 0 0

Cm

ax

(n

g/m

L)

D1

D1

4

D1

D1

4

D1

D1

4

D1

D1

4

D1

D1

4

1 0 m g 2 0 m g 4 0 m g 8 0 m g 1 2 0 m g

0

1 ,0 0 0

2 ,0 0 0

3 ,0 0 0

4 ,0 0 0

5 ,0 0 0

6 ,0 0 0

7 ,0 0 0

AU

C0

-t (

ng

*h

/mL

)

D1

D1

4

D1

D1

4

D1

D1

4

D1

D1

4

D1

D1

4

1 0 m g 2 0 m g 4 0 m g 8 0 m g 1 2 0 m g

HM71224, a Selective BTK Inhibitor, Ameliorates Murine Lupus Development

Yu -Yon Kim1, Ki Tae Park1, Kyu Hang Lee1, Sun Young Jang1, Tae Hun Song1, Young-Mi Lee1, Young Hoon Kim1, Kwee Hyun Suh1 1Hanmi Research Center, Hanmi Pharmaceutical Co. Ltd., Korea, Republic Of

Introduction

FRI0380

Methods

Results

The 16th European League Against Rheumatism (EULAR) Annual European Congress, Rome, Italy; June 10-13, 2015

• Systemic lupus erythematosus (SLE) is known to be associated with the

formation of autoantibodies by B cell hyperactivity.

• Bruton’s tyrosine kinase(BTK) is a member of TEC family tyrosine kinases

important in B cell activation, proliferation, survival and differentiation.

• HM71224 is an orally irreversible BTK inhibitor with IC50 of 2.6 nM in kinase

inhibition assay and IC50 of 1 nM in phosphorylation inhibition assay.

• Commonly used model for SLE drug discovery is the spontaneous lupus mice

model, MRL/lpr with the lymphoproliferation mutation(Faslpr) and New Zealand

Black and White F1 hybrid (NZB/W F1) strain with co-expression of several sle

susceptibility genetic loci.

• The aim of this study is to evaluate the impacts of therapeutic intervention

on the development of SLE like disease features by HM71224 in MRL/lpr

and NZB/W F1 mice lupus models.

Clin Pharmacol Ther. 2012 91(1) 143-9

1. Animals and Administration

8

18

Dosing start

18

28~32

28 weeks old

40 weeks old

Disease onset End of the study

Mice

Female MRL/lpr

NZB/W F1

N=12/group once a day

• The measurements of urine protein with urine strip, blood urea

nitrogen and creatinine with chemical analyzer and anti-dsDNA IgG

with ELISA were conducted.

• Relative organ weights of spleen, cervical lymph node and kidney

were measured.

• Phenotyping of splenic B cells were performed by flowcytometry.

• Renal histopathology was scored in H&E and PAS stain.

• Survival rates were calculated with the Kaplan-Meier method.

• Statistical significance between groups was evaluated by one-way

ANOVA with Dunnett’s test or Kruskal-Wallis with Dunn’s test.

2. Results in MRL/lpr Mice

1. In vitro Cellular Activity

Cell Inhibition of phospho-kinase (IC50, nM)

Stimulant p-Kinase HM71224

Ramos 1) anti-IgM

p-BTKY223 1.0

p-PLCγ2 1.0

IL-4 p-STAT6 445

CTLL-2 2) IL-2 p-STAT5 >1,000

A431 3) EGF p-EGFR 800

1) Human Burkitt's lymphoma B-cell line, 2) murine cytotoxic T-cell line, 3) human epidermoid carcinoma cell line

• HM71224 is a potent and selective BTK inhibitor

• HM71224 prevents the development of skin lesions

V e h ic le 5 0 3 1 0 3 0

0 .0

0 .5

1 .0

1 .5

2 .0

H M 7 1 2 2 4C y c lo p h o s p h a m id e

(m g /k g )

* * * * * * * * * * * *Cli

nic

al

Sc

ore

at

28

we

ek

s * * * p < 0 .0 0 1 v s . v e h ic le Vehicle HM71224 (10mg/kg, QD)

2. Observations V e h ic le 5 0 3 1 0 3 0

0

1 0

2 0

3 0

4 0

* * *

* *

H M 7 1 2 2 4C y c lo p h o s p h a m id e

(m g /k g )

B2

20

+C

D6

9+ (

x1

02 c

ell

s /

10

4c

ell

s)

* * p < 0 .0 1

* * * p < 0 .0 0 1

v s . v e h ic le

V e h ic le 5 0 3 1 0 3 0

0

2 0 0

4 0 0

6 0 0

* * *

H M 7 1 2 2 4C y c lo p h o s p h a m id e

(m g /k g )

An

ti-d

sD

NA

Ig

G (

un

it/u

l)

* * * p < 0 .0 0 1

v s . v e h ic le

V e h ic le 5 0 3 1 0 3 0

0

1 0

2 0

3 0

* * *

H M 7 1 2 2 4C y c lo p h o s p h a m id e

(m g /k g )

B2

20

+G

L7

+ (

x1

02c

ell

s/1

04c

ell

s)

* * * p < 0 .0 0 1

v s . v e h ic le

Spleen Cervical LN Kidney

0.0

0.5

1.0

1.5

2.0 Vehicle control

Cyclophosphamide, 50 mg/kg, ip, QW

HM71224, 3 mg/kg, po, QD

HM71224, 10 mg/kg, po, QD

HM71224, 30 mg/kg, po, QD

***

***

*

***

**

* ** ****

Weig

ht

(g)

* p<0.05, ** p<0.01, *** p<0.001 vs. vehicle

a. Splenic Activated B cells b. Splenic Germinal Center B cells

d. Organ weights c. Autoantibody

• HM71224 inhibits the B cell activation, autoantibody production

and enlargement of lymphatic organs

a. Serum BUN

V e h ic le 5 0 3 1 0 3 0

0

1 0

2 0

3 0

4 0

* * * *

H M 7 1 2 2 4C y c lo p h o s p h a m id e

(m g /k g )

Se

ru

m B

UN

(m

g/d

l)

* p < 0 .0 5

* * * p < 0 .0 0 1

v s . v e h ic le

V e h ic le 5 0 3 1 0 3 0

0

1

2

3

4

* * *

** *

H M 7 1 2 2 4C y c lo p h o s p h a m id e

(m g /k g )

Glo

me

ru

lon

ep

hrit

is S

co

re * p < 0 .0 5

* * p < 0 .0 1

* * * p < 0 .0 0 1

v s . v e h ic le

V e h ic le 5 0 3 1 0 3 0

0

1

2

3

4

* * *

H M 7 1 2 2 4C y c lo p h o s p h a m id e

(m g /k g )

Inte

rs

tit

ial

Infla

mm

atio

n

&F

ibro

sis

Sc

ore

* * * p < 0 .0 0 1

v s . v e h ic le

• HM71224 ameliorates the renal injury and inflammation from SLE

b. GN score c. IN score

3. Results in NZB/W F1 Mice

• HM71224 inhibits the activation of B lymphocytes

d. Survival Rates c. Urine Protein

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2

5 0

6 0

7 0

8 0

9 0

1 0 0

1 1 0

V e h ic le c o n t r o l

M y c o p h e n o la t e

m o fe t i l , 3 0 m g /k g

H M 7 1 2 2 4 , 3 m g /k g

H M 7 1 2 2 4 , 1 0 m g /k g

H M 7 1 2 2 4 , 3 0 m g /k g

W e e k s a fte r A d m in is tra tio n

% S

urv

iva

l Survival rate Vehicle 66.7% MMF (30mpk) 91.7% HM (3mpk) 75.0% HM (10mpk) 100% HM (30mpk) 100%

1 1 1 1 1

0

5

1 0

1 5

M y c o p h e n o la te

m o fe til

H M 7 1 2 2 4

V e h ic le 30 3 10 30

* *

*

B2

20

+C

D6

9+ (

x1

02 c

ell

s /

10

4c

ell

s)

* p < 0 .0 5

* * p < 0 .0 1

v s . v e h ic le

1 1 1 1 1

0

2

4

6

M y c o p h e n o la te

m o fe til

H M 7 1 2 2 4

V e h ic le 30 3 10 30

* * *

*

B2

20

+/C

D1

38

+ (

x1

02c

ell

s/1

04c

ell

s)

* p < 0 .0 5

* * * p < 0 .0 0 1

v s . v e h ic le

a. Splenic Activated B cells b. Splenic Plasma B cells

V e h ic le 3 0 3 1 0 3 0

0

1

2

3

4

H M 7 1 2 2 4M y c o p e n o la te

m o fe til

(m g /k g )

Urin

e p

ro

te

in s

co

re

(4

0 w

ee

ks

of

an

ima

l a

ge

)

* *

** p < 0 .01

v s . v e h ic le

* *

G N1 )

s c o re IN2 )

s c o re V I3 )

s c o re

0

1

2

3

4V e h ic le c o n tro l

H M 7 1 2 2 4 , 3 0 m g /k g , p o , Q D

M y c o p h e n o la te m o fe til,

3 0 m g /k g , ip , Q D

H M 7 1 2 2 4 , 1 0 m g /k g , p o , Q D

H M 7 1 2 2 4 , 3 m g /k g , p o , Q D

*

*

* * * ** * *

His

top

ath

olo

gic

al

Sc

ore

(Me

an

± S

D)

1 ) G lo m e r u lo n e p h r i t i s

2 ) In te r s t i t ia l N e p h r it is

3 ) V a s c u l it is

* p < 0 .0 5 , * * p < 0 .0 1 , * * * p < 0 .0 0 1 v s . v e h ic le

Vehicle

HM71224 3 mg/kg HM71224 10 mg/kg HM71224 30 mg/kg

Mycophenolate mofetil

30 mg/kg

a. Renal Histopathology

• HM71224 reduces renal damages and lymphocyte infiltration

The thickening of basement membrane(arrow) and tubule distended with proteinaceous fluid(asterisk) were ameliorated in HM71224 treated groups.

- Effectively dampened splenic B cells and autoantibody.

- Significantly decreased the development of SLE like

manifestations such as skin lesions, enlargement of

lymph node, splenomegaly, urine protein and renal injury.

- Markedly decreased mortality from SLE in NZB/W F1 mice

model.

• BTK inhibition by HM71224 in MRL/lpr and NZB/W F1 mice

• HM71224 can efficaciously ameliorate lupus developments

in murine disease model that resembles human SLE.

Conclusion

References • Current Protocol in Pharmacology 2011; 5 : 5.60.1-5.60.40.

• BioDrugs 2013; 27(2), 85-95.

• Arthritis Research & Therapy 2012; 14, R243.