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CHITINASE INHIBITORS DEVELOPED BY OAT USED AS TOOLS IN OUR
MOA AND POC STUDIES
ACKNOWLEDGEMENTS
OATD-01 EFFICACY IN BLEOMYCIN-INDUCED PULMONARY FIBROSIS
MODEL IN MICE
CHRONIC HDM-INDUCED AIRWAY INFLAMMATION AND
REMODELING MODEL
INTRODUCTION
Acidic mammalian chitinase (AMCase) and chitotriosidase (CHIT1) are enzymatically active chitinases that have been implicated in
the pathology of chronic lung diseases. Significantly elevated chitinolytic activity was demonstrated in asthma, chronic obstructive
pulmonary disease (COPD) and interstitial lung diseases such as idiopathic pulmonary fibrosis (IPF) and sarcoidosis.
Herein, we describe our studies on targeting chitinases with small molecules as a potential therapy for pulmonary diseases.
PHARMACOKINETIC PROFILE IN MICE
OATD-01 has a favorable pharmacokinetics in mice suitable for once- or twice-a-day dosing regimen in mice.
CONCLUSIONS
✓ OATD-01 is the first-in-class chitinase inhibitor to have entered clinical trials (currently in phase 1b).
✓ OATD-01 is a nanomolar inhibitor of AMCase and CHIT1 with optimal pharmacokinetic profile in rodents and dogs.
✓ In addition to anti-inflammatory efficacy in asthma models, OATD-01 exhibited anti-fibrotic effects in a chronic HDM-
induced airway remodelling model and in a bleomycin-induced pulmonary fibrosis model in mice.
These data indicate that inhibition of chitinases might represent a novel therapeutic approach for pulmonary diseases as
well as several fibrotic pathologies.
OATD-01, A Dual hAMCase and hCHIT Inhibitor as a Potential Therapeutic Agent for Treatment of Pulmonary Diseases.Agnieszka Bartoszewicz1, Szymon Kłossowski1, Marzena Mazur1, Sylwia Olejniczak1, Robert Koralewski1, Michał Kowalski1, Barbara Dymek1, Piotr Sklepkiewicz1, Michał Mlącki1, Wojciech Czestkowski1, Gleb
Andryianau1, Elżbieta Pluta1, Piotr Niedziejko1, Krzysztof Matyszewski1, Mariusz M. Gruza1, Łukasz Joachimiak, Agnieszka Zagożdżon1, Magdalena Salamon1, Jakub Gołab2, Marcin Nowotny3,
Agnieszka Napiórkowska3, Małgorzata Kwiecień3, Karolina Dzwonek1, Paweł Dobrzański1, Jacek Olczak1, Adam Golebiowski1
1OncoArendi Therapeutics SA, Żwirki i Wigury 101, 02-089 Warsaw, Poland; 2Department of Immunology, Medical University of Warsaw, Nielubowicza 5, 02-097 Warsaw, Poland3ProBiostructures, International Institute of Molecular and Cell Biology, Trojdena 4, 02-109 Warsaw, Poland.
REFERENCES
* * * *
No
rmal co
ntr
ol
Sarc
oid
osis
IPF
CO
PD
0
1
2
3
4
5
6
Ch
itin
oly
tic
ac
tiv
ity
[µM
/µl/
h]
* * * *
* *
S E R U M
No
rmal co
ntr
ol
Sarc
oid
osis
IPF
CO
PD
0 .0
0 .2
0 .4
0 .6
1 .0
1 .5
2 .0
2 .5
3 .0
Ch
itin
oly
tic
ac
tiv
ity
[µM
/µl/
h]
*
* * *
* *
IN D U C E D S P U T U M
Sarc
oid
osis
IPF
CO
PD
0 .0
0 .1
0 .2
0 .3
0 .4
0 .5
0 .5
1 .0
1 .5
2 .0
Ch
itin
oly
tic
ac
tiv
ity
[µM
/µl/
h]
B A L F
SAR STUDYIn the course of our program over 2500 compounds have been designed and synthesized, resulting in OAT-870 as our advanced
lead compound. Further optimization of drug-like properties and selectivity of OAT-870 yielded a clinical candidate OATD-01.
The compound bears an additional methyl group at the morpholine ring, which abrogated undesired off-target activity towards
dopamine transporter (DAT).5,6
„Preclinical research and clinical trials of a first-in-class development candidate in therapy of asthma and inflammatory
bowel disease”
hAMCase IC50 = 14 nM
hCHIT1 IC50 = 232 nM
mAMCase IC50 = 19 nM
mCHIT1 IC50 = 2955 nM
17x
155x
Selective inhibitor of mAMCase2
hAMCase IC50 = 4.7 nM
hCHIT1 IC50 = 830 nM
Selective inhibitor of hAMCase3
mCHIT1 IC50 = 29 nM
mAMCase IC50 = 4170 nM
Selective inhibitor of mCHIT4
143x176x
Poster MEDI69
OATD-01 Pharmacokinetic Parameters
Route IV PO
Dose (mg/kg) 3 10
AUC0-inf(mg·h/L) 8.6 22.2
AUCn0-inf(kg·h/L) 2.9 2.2
C0 or Cmax(mg/L) 4.0 3.4
Tmax(h) n/a 2.0
CL (mL/min/kg) 5.8 n/a
Vss (L/kg) 1.0 n/a
T½ (h) 2.1 1.9
Bioavailability (F%) n/a 77
CRYSTAL STRUCTURE OF OATD-01-CHIT1 COMPLEX
✓ Treatment with OATD-01 significantly decreased chitinolytic activity in BAL indicating target engagement in the lungs in vivo.
Decrease in chitinolytic activity correlated with reduction of either collagen level in the lungs or edema represented by lung
weight to body weight ratio
A) Stereoview of key residues (shown as green sticks) of hCHIT1
active site. Protein−ligand hydrogen bonds are represented by orange dashed lines and distances are given in Å.
B) Active site pocket of hCHIT1 represented as surface and
colored according to electrostatic potential.
mRNA profile of chitinases in IPF lungs
0 .0
0 .2
0 .4
0 .6
0 .8
Ch
itin
oly
tic
ac
tiv
ity
in
pla
sm
a,
pH
6,
[µM
/µl/
h]
C o n tro l
B le o m y c in (2 U /k g )
O A T D -0 1 (3 0 m g /k g b . i.d )
P ir fe n id o n e (2 5 0 m g /k g b . i.d )
* * * * * * * *
Day 0 7 21
Bleomycin 2U/kg,
3x i.n. (d0-2)OATD-01 (30mg/kg; PO; b.i.d)
PIRFENIDONE (250mg/kg; PO; b.i.d)
0
5 0
1 0 0
1 5 0
So
lub
le c
oll
ag
en
[m
g/l
un
gs
]
* * *
C o n tro l
B le o m y c in (2 U /k g )
O A T D -0 1 (3 0 m g /k g b . i.d )
P ir fe n id o n e (2 5 0 m g /k g b . i.d )
0
1 0
2 0
3 0
Lu
ng
/Bo
dy
we
igh
t r
atio
C o n tro l
B le o m y c in (2 U /k g )
O A T D -0 1 (3 0 m g /k g b . i.d )
P ir fe n id o n e (2 5 0 m g /k g b . i.d )
* * * *
OATD-01 reduced the goblet cell hyperplasia as assessed with periodic acid-Shiff (PAS) staining of mucins in mouse lung tissue.
1) Yang IV et al. Thorax. 2013, 68, 1114, Data extracted from the study GSE32537 (IPF n=119; Control n=50)
2) Mazur, M.; Olczak, J.; Olejniczak, S.; Koralewski, R.; Czestkowski, W.; et al. J. Med. Chem. 2018, 61, 695.
3) Mazur M. et al. US20170066743
4) Mazur M.; Bartoszewicz A.; Dymek B.; Salamon, M.; Adrianau, G.; et al. Bioorg. Med. Chem. Lett., 2018, 28, 310.
5) Mazur, M.; Dymek, B.; Koralewski, R.; Sklepkiewicz, P.; Olejniczak, S.; et. al. J. Med. Chem. 2019, ASAP
6) Koralewski R.; Dymek, B.; Mazur, M.; Sklepkiewicz, P.; Olejniczak, S.; et. al. submited
Poster MEDI67
hAMCase IC50 = 24 nM
hCHIT1 IC50 = 47 nM
mAMCase IC50 = 20 nM
mCHIT1 IC50 = 77 nM
DAT 94% inhibition at 10 mM
hAMCase IC50 = 54 nM
hCHIT1 IC50 = 14 nM
mAMCase IC50 = 1 nM
mCHIT1 IC50 = 14 nM
hAMCase IC50 = 3 nM
hCHIT1 IC50 = 18 nM
mAMCase IC50 = 2 nM
mCHIT1 IC50 = 19 nM
DAT 34% inhibition at 10 mM
hAMCase IC50 = 23 nM
hCHIT1 IC50 = 52 nM
mAMCase IC50 = 111 nM
mCHIT1 IC50 = 87 nM
hAMCase IC50 = 826 nM
hCHIT1 IC50 = 5100 nM
mAMCase IC50 = 1000 nM
mCHIT1 IC50 = 6400 nM
hAMCase IC50 = 9 nM
hCHIT1 IC50 = 23 nM
mAMCase IC50 = 8 nM
mCHIT1 IC50 = 28 nM
DAT IA
C57Bl/6
✓ In 7-week-long HDM-induced airway
inflammation model OATD-01 administered
qd in a therapeutic regiment significantly
reduced the total number of leukocytes
and eosinophils in BALF.
✓ Anti-inflammatory activity of OATD-01
correlates with a significantly reduced
chitinolytic activity in BALF and plasma.
✓ OATD-01 reduced the serum IgE
concentration in a dose-dependent manner
indicating reduction of allergic response.
OATD-01(30mg/kg)
OATD-01 30mg/kg
OATD-01
OATD-01
OATD-01 exhibited significant anti-remodeling activity: decreased airway wall thickness and reduced collagen deposition around
bronchioles.
0
1
2
3
4
PA
S s
tain
ing
me
an
sc
ore *
C o n tro l
C h ro n ic H D M
O A T D -0 1 (3 m g /k g q d )
O A T D -0 1 (3 0 m g /k g q d )
* * * *
HDMControl
OATD-01 30mg/kgHDMControl
0
2
4
6
Ch
itin
oly
tic
ac
tiv
ity
in
BA
L,
pH
6
[ µM
/µl/
h]
(me
an
wit
h S
EM
)
***
Control
HDM
OATD-01 (3 mg/kg)
OATD-01 (30 mg/kg)
Dexamethasone (10 mg/kg)
0
50000
100000
150000
To
tal
nu
mb
er
of
BA
L e
os
ino
ph
ils
(me
an
wit
h
SE
M)
Control
HDM
OATD-01 (3 mg/kg)
OATD-01 (30 mg/kg)
Dexamethasone (10 mg/kg)
***
0
10000
20000
30000
40000
50000
60000
To
tal
IgE
in
pla
sm
a
[ng
/ml]
Control
HDM
OATD-01 (3 mg/kg)
OATD-01 (30 mg/kg)
Dexamethasone (10 mg/kg)
0
50000
100000
150000
200000
250000
To
tal B
AL
(C
D4
5+)
ce
ll n
um
be
r
(me
an
wit
h S
EM
)
Control
HDM
OATD-01 (3 mg/kg)
OATD-01 (30 mg/kg)
Dexamethasone (10 mg/kg)
***
Control Bleomycin (2U/kg) OATD-01 (30mg/kg, b.i.d) Pirfenidone (250mg/kg; b.i.d)
✓ OATD-01 treatment decreases pulmonary fibrosis formation in bleomycin induced fibrosis model.
Contr
ol
Ble
omyc
in
OATD
-01
(30
mg/k
g bid
)
Pirfen
idone
(250
mg/k
g bid
)0
2
4
6
8
Ash
cro
ft fi
bro
sis
sco
re
*****
*