2DG - a glucose analogue and glycolytic inhibitor with anticonvulsant and antiepileptic actions Tom Sutula, MD, PhD Department of Neurology, University of Wisconsin Chief Scientific Officer, NeuroGenomex, Inc.

cannot undergo isomerization The novel mechanism of 2DG: why 2DG is a glycolytic inhibitor

Completed preclinical efficacy studies Animal Models of Acute & Chronic Epilepsy in vitro Models of Seizure Induction  protection against seizures evoked by 6Hz stimulation  protection against audiogenic seizures in Fring’s mice  2-fold slowing of latency to status epilepticus onset by kainic acid, pilo  2-fold slowing of kindling progression evoked from different brain sites  time course of action & dose range  effective against seizure progression when administered up to 10 minutes after a seizure 2DG reduces epileptic discharges evoked by:  K + (7.5mM, ictal & interictal)  bicuculline (GABA antagonist)  4AP (K + channel antagonist)  DHPG (metabotropic glutamate agonist )

Completed preclinical efficacy studies Animal Models of Acute & Chronic Epilepsy in vitro Models of Seizure Induction  these properties are unlike ANY other marketed drugs  2DG is disease- modifying against progressive adverse effects of seizures  efficacy studies support clinical utility for both ACUTE and CHRONIC uses 2DG reduces epileptic discharges evoked by:  K + (7.5mM, ictal & interictal)  bicuculline (GABA antagonist)  4AP (K + channel antagonist)  DHPG (metabotropic glutamate agonist )  protection against seizures evoked by 6Hz stimulation  protection against audiogenic seizures in Fring’s mice  2-fold slowing of latency to status epilepticus onset by kainic acid, pilo  2-fold slowing of kindling progression evoked from different brain sites  time course of action & dose range  effective against seizure progression when administered up to 10 minutes after a seizure  implies that actions of 2DG at the cellular level are “broad- spectrum” against different mechanisms of network synchronization

Completed preclinical efficacy studies Animal Models of Acute & Chronic Epilepsy in vitro Models of Seizure Induction  implies that actions of 2DG at the cellular level are “broad- spectrum” against different mechanisms of network synchronization  these properties are unlike ANY other marketed drugs  2DG is disease- modifying against progressive adverse effects of seizures  efficacy studies support clinical utility for both ACUTE and CHRONIC uses 2DG reduces epileptic discharges evoked by:  K + (7.5mM, ictal & interictal)  bicuculline (GABA antagonist)  4AP (K + channel antagonist)  DHPG (metabotropic glutamate agonist )  protection against seizures evoked by 6Hz stimulation  protection against audiogenic seizures in Fring’s mice  2-fold slowing of latency to status epilepticus onset by kainic acid, pilo  2-fold slowing of kindling progression evoked from different brain sites  time course of action & dose range  effective against seizure progression when administered up to 10 minutes after a seizure

Completed preclinical efficacy studies Animal Models of Acute & Chronic Epilepsy in vitro Models of Seizure Induction  implies that actions of 2DG at the cellular level are “broad- spectrum” against different mechanisms of network synchronization  these properties are unlike ANY other marketed drugs  2DG is disease- modifying against progressive adverse effects of seizures  efficacy studies support clinical utility for both ACUTE and CHRONIC uses 2DG reduces epileptic discharges evoked by:  K + (7.5mM, ictal & interictal)  bicuculline (GABA antagonist)  4AP (K + channel antagonist)  DHPG (metabotropic glutamate agonist )  protection against seizures evoked by 6Hz stimulation  protection against audiogenic seizures in Fring’s mice  2-fold slowing of latency to status epilepticus onset by kainic acid, pilo  2-fold slowing of kindling progression evoked from different brain sites  time course of action & dose range  effective against seizure progression when administered up to 10 minutes after a seizure

Completed preclinical efficacy studies Animal Models of Acute & Chronic Epilepsy in vitro Models of Seizure Induction  implies that actions of 2DG at the cellular level are “broad- spectrum” against different mechanisms of network synchronization  these properties are unlike ANY other marketed drugs  2DG is disease- modifying against progressive adverse effects of seizures  efficacy studies support clinical utility for both ACUTE and CHRONIC uses 2DG reduces epileptic discharges evoked by:  K + (7.5mM, ictal & interictal)  bicuculline (GABA antagonist)  4AP (K + channel antagonist)  DHPG (metabotropic glutamate agonist )  protection against seizures evoked by 6Hz stimulation  protection against audiogenic seizures in Fring’s mice  2-fold slowing of latency to status epilepticus onset by kainic acid, pilo  2-fold slowing of kindling progression evoked from different brain sites  time course of action & dose range  effective against seizure progression when administered up to 10 minutes after a seizure

likely useful for acute epileptic conditons with ongoing seizures (eg., status epilepticus, clusters) potentially useful for peri- seizure administration to optimize delivery and minimize side-effects potentially novel methods of delivery in combination with stimulation /device therapies Implications for clinical applications 2DG delivery to neurons and circuits is activity-dependent: a novel advantageous property for an anticonvulsant 18F-2DG PET scan during status epilepticus H3-2DG autoradiogram during 1 hz stimulation 2DG is preferentially delivered to regions with high energy needs by activity-dependent neurovascular coupling involving neurons, glia, and endothelial cells in the “neurovascular unit”

likely useful for acute epileptic conditons with ongoing seizures (eg., status epilepticus, clusters) potentially useful for peri- seizure administration to optimize delivery and minimize side-effects potentially novel methods of delivery in combination with stimulation /device therapies Implications for clinical applications 2DG delivery to neurons and circuits is activity-dependent: a novel advantageous property for an anticonvulsant 18F-2DG PET scan during status epilepticus H3-2DG autoradiogram during 1 hz stimulation 2DG is preferentially delivered to regions with high energy needs by activity-dependent neurovascular coupling involving neurons, glia, and endothelial cells in the “neurovascular unit”

likely useful for acute epileptic conditons with ongoing seizures (eg., status epilepticus, clusters) potentially useful for peri- seizure administration to optimize delivery and minimize side-effects potentially novel methods of delivery in combination with stimulation /device therapies Implications for clinical applications 2DG delivery to neurons and circuits is activity-dependent: a novel advantageous property for an anticonvulsant 18F-2DG PET scan during status epilepticus H3-2DG autoradiogram during 1 hz stimulation 2DG is preferentially delivered to regions with high energy needs by activity-dependent neurovascular coupling involving neurons, glia, and endothelial cells in the “neurovascular unit”

likely useful for acute epileptic conditons with ongoing seizures (eg., status epilepticus, clusters) potentially useful for peri- seizure administration to optimize delivery and minimize side-effects potentially novel methods of delivery in combination with stimulation /device therapies Implications for clinical applications 2DG delivery to neurons and circuits is activity-dependent: a novel advantageous property for an anticonvulsant 18F-2DG PET scan during status epilepticus H3-2DG autoradiogram during 1 hz stimulation 2DG is preferentially delivered to regions with high energy needs by activity-dependent neurovascular coupling involving neurons, glia, and endothelial cells in the “neurovascular unit”

Activity-dependent effects of 2DG on synaptic transmission

Activity-dependent effects of 2DG on synaptic transmission no effects on synaptic properties in normal conditions

2DG reduces amplitude and frequency of sEPSCs after exposure to conditions that increase activity Activity-dependent effects of 2DG on synaptic transmission

modification in mEPSCs is activity-dependent implicates presynaptic vesicle release as a mechanism of action of 2DG Activity-dependent effects of 2DG on synaptic transmission NOT SHOWN: “homeostatic” effects on synaptic plasticity

2DG in the pipeline  PET imaging agent for 30+ years  Favorable toxicity history Completed Phase 1 in cancer More than 20 other investigator clinical studies with > 300 subjects Need to confirm safety in effective dose (ED) range – cardiac toxicity observed with chronic dosing at 10X > ED range, and mild potentially reversible cardiac autophagy in ED range  bioanalytical assay development, toxicity, PK, TK, ADME studies underway in 2010 Q2 Toxicology  distinctive pattern of effectiveness in pre-clinical models  disease-modifying against progressive adverse effects of seizures  novel acute and chronic anti- epilepsy mechanisms of action  2DG concentrates in areas of ictal epileptic activity Efficacy  IP licensed from WARF: 1 patent issued and 1 pending in US (has been issued in Australia)  NGX has exclusive license from WARF for all human therapeutic use Intellectual Property Remaining preclincal studies

2DG in the pipeline  PET imaging agent for 30+ years  Favorable toxicity history Completed Phase 1 in cancer More than 20 other investigator clinical studies with > 300 subjects Need to confirm safety in effective dose (ED) range – cardiac toxicity observed with chronic dosing at 10X > ED range, and mild potentially reversible cardiac autophagy in ED range  bioanalytical assay development, toxicity, PK, TK, ADME studies underway in 2010 Q2 Toxicology  distinctive pattern of effectiveness in pre-clinical models  disease-modifying against progressive adverse effects of seizures  novel acute and chronic anti- epilepsy mechanisms of action  2DG concentrates in areas of ictal epileptic activity Efficacy  IP licensed from WARF: 1 patent issued and 1 pending in US (has been issued in Australia)  NGX has exclusive license from WARF for all human therapeutic use Intellectual Property Remaining preclincal studies IND filing anticipated 2011 Q1

Current Development Plan Complete preclinical toxicity, formulation, CMC, and filing of IND “IND-enabling preclinical studies of 2DG for treatment of epilepsy” (awaiting NIH RAID, anticipated start Q2 2010) Will complete preclinical studies including bioanalytical assay development, pharmacokinetic, toxicological, toxicokinetic, manufacturing, formulation, and clinical trial designs, and regulatory documentation for submission of an IND. Investigator-initiated first in humans Phase I/II clinical trial in patients with intractable epilepsy “A Preliminary Tolerability and Efficacy Study of 2DG in Intractable Epilepsy” Nathan Fountain, MD, University of Virginia (EpilepsyTherapy Project-ERF, WARF, NGX, anticipated start in Q4 2010) This will be a preliminary study of 2DG that will seek to detect an efficacy signal and assess tolerability in intractable patients with frequent seizures. Development of delayed release formulations Zeeh Experimental Pharmacy Station (University of Wisconsin) (E. Elder, PhD, supported by WARF) This program is developing delayed release formulations to exploit the activity-dependent uptake and short t 1/2 (~ 40 min) enabling chronic administration at lower total doses.