GABAA Receptor Modulators for Treatment of Bronchoconstrictive Disorders Novel Target and Compounds

Inventors

Doug Stafford, PhD Charles Emala, MDJames Cook, PhD George Gallos, MDAlexander Arnold, PhD

Licensing Officer

Sara GusikColumbia Technology VenturesColumbia Universitysag23@Columbia.edu

Executive Summary

Novel target for treatment of bronchoconstritivediseases and library of GABA subunit selective compounds 3 compounds are confirmed GABAA targeting agents with the capacity to relax airway smooth muscle with specificity and attenuate the inflammatory response.

Technology

Asthma

Chronic Obstructive Pulmonary Disease ( COPD)Target

Markets

Intellectual Property

2

Patent Application ( pending US, EP, Japan and Canada)Novel GABAA Agonists and Methods of Using to Control Airway Hyperresponsiveness and inflammation in Asthma (WO2014/047413)

Target Markets

AsthmaAmong adult asthma suffers, the greatest increase will be in adults over 65 years of age.2

COPDWhile historically underserved and underdiagnosed, COPD market is seeing high growth momentum. 2

41.7 MillionPrevalence of

Asthma in the US 1

16.8 MillionCOPD patient

population in US 20161

1Frost & Sullivan 2014 Competitive Analysis of Asthma and COPD Therapeutics 3

Total Global Market for Asthma is expected to reach $16.2 Billion in 2022

The Market demands innovative drug compositions for Asthma and COPD, current therapies in development are modifications of the same drugs/ steroids

GABAA Agonist Demonstrates Site and Subtype Selectivity

Published data has confirmed that human airway smooth muscle (ASM) cells express limited GABAAR subunits, with the only α subunits being α4 and α 5. Targeting GABA receptors is a proven and safe pharmaceutical approach.

Target selective towards the α subunit in ASM cells limits CNS effects

Immunohistochemistry of Human ASM shows expression of GABAAR α5 subunit

4Li, X. et al.. 2002. Med. Chem. Res. 11(9):504-537.

α4 specific

Compoundα1 α2 α3 α4 α5 α6

XHEIII-74 77 105.5 38.5 0.42 22 5.8

CMD-45 90.5 65.5 30.3 0.15 1.65 0.23

In vitro binding affinity of the GABAA agonist to GABAA/Benzodiazepine site subtypes

The GABAA agonist was assayed using two electrode voltage clamp measurements on Xenopus oocytes expressing single α-subtypes. Functional selectivity to α4 subtype is demonstrated by dose-response curve showing greater inward current(agonist activity)

GABAA α 4 and α5 Subtype Selective Agents Result in ASM Relaxation

Novel compounds targeting GABAAR α4 and α5 subunits induce ASM relaxation 5

Gallos,G . Et al. 2015. Am. J. Phys. 308(9):L931-L942

Agonists induced dose-dependent relaxation of guinea pigtracheal rings contracted with depolarizing stimulus TEA.$$, compared to initial tone, **p<0.01,***p<0.001,compared toCM-D45

GABAA α4 selective agents mediate human ASM relaxationpost acetylcholine-induced muscle contraction. * p<0.05

Representative tracings demonstrate spontaneous relaxation followingaddition of α5 selective agent SH-053 as compared to vehicle control inTEA contracted samples.

GABAA α5 selective agent significantly relaxes TEA inducedmuscle force (tension) 15 minutes after addition. * p<0.05

GABAA α5 Selective Compound’s Mechanism of Action

Ex-Vivo studies have provided insights into the mechanism of action of the GABAA α5 receptor selective agents and ASM relaxation.

GABAA α5 selective agent relaxes peripheral murine airway slices pre-contracted with methacholine (MCh). Representative phase contrast images of airway murine airway slices (1) before stimulation with Mch, (2) after stimulation with MCh (3) after addition of the a5 agonist in the continued presence of MCh and (4) after washout of agonist and continued presence of MCh. ***p<0.001

GABAA α5 receptor activation can directly relax pre-contracted airway smooth muscle by attenuating Ca2+ handling 6

Gallos, G et al. 2015. AJ P lung Phys. epub.

Compiled data from replicates demonstrates significant reduction in Ca2+ oscillations stimulated by MCh with the addition of the GABAA α5 selective agents.***p<0.001

GABAA Selective Compounds Modulate Inflammatory Response

Lung inflammation is a hallmark of asthma, genetic deletion of GABAA α4 leads to increased lung inflammation in an allergic asthma model.

H&E-stained lung sections from house dust mite sensitized (A) WT and (B) GABAA α4 KO mice demonstrate enhanced inflammatory infiltration in the α4 KO mice; quantification by Composite Lung Inflammation Score is shown in the graph

GABAA selective agents can effectively target ASM and inflammatory cells, the principal cell types involved in asthma

7

A B

GABAA agonists significantly reduce lymphocyte proliferation. WT mouse CD4+ cells were stimulated for 72 hours with anti-CD3/CD28 coated beads then co-cultured with a GABAaR agonist Muscimol and GABAA α4 selective compound( Xhe-III-74).

Limitations of Current Treatment Options for AsthmaUnmet need for non-steroidal treatments for bronchoconstructive disorders

8

Limitations of Current Asthma Drugs Advantages of GABAA Targeting Agents

X Inhaled therapies are difficult to administer in some patient populations

X Corticosteroids take several weeks to become effective

X Drug benefits wear off quickly without continued daily use

X Side effects associated with use of steroid drugs

X First line therapies do not satisfactorily treat all patients

Selective for specific GABAAR subunits

Targeting GABAA subsets reduces

potential for CNS effects

Reduced side effects as compared to

steroid therapies

Potential for variable routes of delivery

Attenuates Inflammatory response in

asthma as well as relaxation of smooth

muscle

Limitations in current products demonstrates a need for novel targets and mechanism of action

Strategic Path ForwardAsthma Drug Development

9

UWMRF Catalyst Program (compound screening and animal studies) : $ 50K

NIH/NIHLB ( Preclinical POC studies) : $2M

NIH (MOA in muscle and immune cells): $1.5M Funding

Partnering

Preclinical Proof of Concept

IND enabling studies

First-in-man studies

University of Wisconsin- Milwaukee• Synthetic chemistry, lead optimization and preclinical validation

Medical College of Wisconsin• Asthma models and immunobiology

Columbia University• Respiratory physiology and animal models

Research Team