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

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S E R U M

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IN D U C E D S P U T U M

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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

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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 )

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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)

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B le o m y c in (2 U /k g )

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P ir fe n id o n e (2 5 0 m g /k g b . i.d )

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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.

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O A T D -0 1 (3 0 m g /k g q d )

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HDMControl

OATD-01 30mg/kgHDMControl

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OATD-01 (30 mg/kg)

Dexamethasone (10 mg/kg)

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OATD-01 (30 mg/kg)

Dexamethasone (10 mg/kg)

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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

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