Palisade Bio, Inc. · NASDAQ:PALI
Palisade Bio — Neuralstem — Technical Presentation (May 2017)
30 slides from Palisade Bio's technical presentation, May 31, 2017
Slide 1: NEURALSTEM INC
NEURALSTEM INC “The neurogenic antidepressant compound, NSI-189, shows potential as a broad neurotrophic agent” Karl Johe, Ph.D. Chief Scientific Officer NEURALSTEM, INC.
Slide 2: Safe Harbor & Conflict of Interest Statements
Safe Harbor & Conflict of Interest Statements ‘Safe Harbor statements under the Private Secures Litigation Reform Act of 1995: This presentation contains forward-looking statements as defined in Section 27A of the ‘Secutties Act of 1933 as amended, and section 21E of the Securities Exchange Act of 1
Slide 3: Molecular Psychiatry
Molecular Psychiatry Figure 2 ‘Subcortical volume differences separately for first episode and recurrent MDD patients, versus controls aS tev WS, Accumbens Amygdala”® Hippocampus* subcortical volume differences separately for early and late onset MDD patients, versus controls S Late onset Early onse
Slide 4: Human Hippocampal Neurogenesis in vitro Platform Enables Neurogenic Drug Discovery
Human Hippocampal (HI) Neurogenesis in vitro Platform Enables Neuragenic Drug Discovery +Mitogen - Mitogen for 7 Days Neural Stem Cells (Red)/ Neurons (Green)/ Nuclei (Blue) Glia (Red)/ Nuclei (Blue)
Slide 5: Library Selection: CNS-Active Compounds
Library Selection: CNS-Active Compounds * Libraries selected to target neurogenesis — Kinases & phosphatases — Nuclear receptors — Peptide mimetics — GPCRs * Five structural libraries chosen for diversity (scaffolds) * Selected “2000 compounds per library — Predict in advance for CNS-Availability —
Slide 6: Compound-Induced Neurogenesis
Compound-Induced Neurogenesis Vehicle Control NSI-106
Slide 7: Primary Screen and Additional Filters
Primary Screen and Additional Filters Screened 10,269 Compounds in Primary Screen Re-tested 1,106 Preliminary Hits in Quantitative Screen fe Secondary Hits Proliferation Hits* Neuron-Ratio Hits* >30% >18%
Slide 8: Leads Increase Neuron Number in a 7-day Neurogenesis Assay
Leads Increase Neuron Number in a 7-day Neurogenesis Assay —— NSI-137 NSI-158 ——NSI-150 Ky —#+ NSI-189 H NSI-144 Number of Neurons (% control) 1000 1000000 1.0x10°
Slide 9: Effect of Antidepressants on Primary Stem Cell Neurogenesis Screen
Effect of Antidepressants on Primary Stem Cell Neurogenesis Screen sges eo 8 8 8 8 A. Fluoxetine 400 B, Imipramine Avg. Number Neurons 102 10° 104 49" 10210" 108 108 pM pM 500( C. Rolipram £ © 400 a 3 300 o 3200 5 hoo z ob & 1040" 104 102 10% 104 108 108 107 pM
Slide 10: Leads Affect Multiple Potential Sites of Action During Stages of Neurogenesis
Leads Affect Multiple Potential Sites of Action During Stages of Neurogenesis ( 1. Proliferation 2. Differentiation © 3. Maturation/Function T 1 eer | 1 Committed 1 Immature Hippocampal Mature Neurons eae Neuronal Progerjtor Neurons Neural Stem Cells i } NSI-127, 130, 137, 143, NSI-106, 127, 144, ~
Slide 11: in vivo Neurogenesis Assay: Protocol
in vivo Neurogenesis Assay: Protocol mpd 10 mg/kg, po BrdU Omelet 2 3 45 67 8 9 10 Perf 1) Perfuse & Remove Brain on Day 11 Dentate Gyrus 2) Do coronal sections (30um) 3) Immuno-stain BrdU-labeled cells 4) Count BrdU+ cells in 10 sections/dentate gyrus/mouse 5) Stain for neuronal phenotype Pictures
Slide 12: Leads (10 mg/kg) Stimulate Neurogenesis in Healthy Adult Mouse
Leads (10 mg/kg) Stimulate Neurogenesis in Healthy Adult Mouse NSLLIU *Conducted by Cerep & NeuroDetective Inc.
Slide 13: Leads (10 mg/kg) show significant neurogenic effects
Leads (10 mg/kg) show significant neurogenic effects in healthy adult mouse dentate gyrus 1 Vehicle (n=7) 1=Vehiole (n=6) —-7= NS/-143 (n=8) 2 = NSI-160 (n=8) 2=NSI150(n=7) = NS/-127 (n=8) 3 ME NSH158 (0-7) 3=NSI444 (n=8) —-9= NSI-190 (n=7) 4 MM NSI-189 (n=6) 5 mmm NSI-149 (n=6) + ISI-130 (n=8) 10=
Slide 14: Leads show anti-depression activity in models of depression
Leads show anti-depression activity in models of depression: chronic novelty suppressed feeding mm Water EE Imipramine 60 mg/kg} * peo.as * H Latency to eat (sec) g 500 * pc0105 400 * Latency to eat (sec) g Vehicle NSI-144 NSI-160NSI-158 NSI-189 N=15-20/group; Conducted by PsychoGenics, Inc.
Slide 15: Latency to eat (sec)
1200 1050 Latency to eat (sec) 0 NSI-189 (10-100 mg/kg) and imipramine significantly decreased the latency to eat compared to vehicle (water) after 28 days of oral dosing. * No significant treatment effects on either body weight or neurological observation were seen, *N=20/group, conducted by Psycho
Slide 16: Increases neurogenesis and HI volume
Increases neurogenesis and HI volume BrdU Cell Count DG Volume Total Celt Counts (o=7lgroup) ~ 888888 Volume(mn?) ETI, . og é 28 4 s Volume(ma) Volume(mr?) hed CLP bd 28d p.0.; Histology conducted . NeuroDigiTech
Slide 17: Screening Path to NSI-189
Screening Path to NSI-189 10,269 Small Molecule Compounds through Primary Screen & 16 Neurogenic Compounds in vitro eS 16 Tested for Acute Tox in Mice & 15 Tested for Neurogenesis in Healthy, Adult Mice & 7 Orally Active Neurogenic Leads (3 Structural Classes) 1 Development Candidate Selected NSI-18
Slide 18: A Phase 1B, Randomized, Double-Blind, Placebo-Controlled Study of NSI-189 in MDD
A Phase 1B, Randomized, Double-Blind, Placebo- Controlled, Multiple-Dose Escalation Study Evaluating the Effects of NSI-189 Phosphate, a Neurogenic Compound, in Patients with Major Depressive Disorder (MDD) Maurizio Fava, M.D., Karl Johe, Ph.D., Lev G. Gertsik, MD, Larry Ereshefsky, PharmD, Bettina
Slide 19: Phase 1b Study Design
Phase 1b Study Design * Double-blind, randomized, placebo-controlled, multiple-dose study with three ascending cohorts (N=24) Cohort 1 N=8 (6 drug, 2 placebo) 40 mg QD Cohort 2 N=8 (6 drug, 2 placebo) 40 mg BID Cohort3 1-8 (6 drug, 2 placebo) 40 mg TID Drug Treatment: 28 days | No-Drug Follow up: 35
Slide 20: Positive Phase 1b clinical outcomes
Positive Phase 1b clinical outcomes MADRS ‘and Asberg CPFQ ‘Gogritve and Physical Rating Scale 9-009 60.95 0.02 60.90 aise Ta NS pray 9 ne 82 pera) Sn. perey 90.03] 1.10} Ey 100 Study Day vas 155
Slide 21: Positive biomarker results
Positive biomarker results qEEG Topographs of High Frequency alpha (10-12 Hz): Day 28 from Baseline NSI-189 Placebo Pre-dose Post-dose Change from Baseline 100 40° 150: Left Hippocampal Vol Study Days Placebo Left posterior temporal (T5) (t=2.45, p=0.02) Left parietal regions (P3) (t=3.31, p=0.004)
Slide 22: NSI-189 produces a concentration-dependent enhancement of LTP magnitude
NSI-189 produces a concentration-dependent enhancement of LTP magnitude 400, © Control - 10uM -# 3uM cy ~ £ 1UM © 300M -& 100nM 2 — 3 : g 8 200 * 2 5 46 s s o © 100 Qa Qa ° 2 Bo 5 o o 9 2 0 o ui issgsgsee u gers s 40 5 0 5 10 1% 2 2 90 35 40 Time (min)
Slide 23: NSI-189 produces a time-dependent enhancement of LTP magnitude
NSI-189 produces a time-dependent enhancement of LTP magnitude 3 -© Control -© NSI-189 (2.5 h) 3 NSI-189 (3.5 h) 8 3 50. 3 fEPSP slope (% of baseline) fEPSP slope (% of baseline) a § 0 5 10 15 20 2 30 35 40 37 Time (min)
Slide 24: NSI-189 restores LTP in hippocampal slices from Angelman Syndrome model
NSI-189 restores LTP in hippocampal slices from Angelman Syndrome model = 20 ¢ > WT © AS -# AS#NSI-189 o = e # 5 & 200. tee :} 8 2 150. s ro ® s a . 100: 2 a . 2 60 G & o oa 0 UY » u $ ° ¢ 40 6 0 5 0 15 2 2% 30 35 40 * Time (min) Yan Liut, Michael P. Hefferan2, Karl Johe2, Xiaoning Bi1, Michel Baudr
Slide 25: NSI-189 ameliorates cognitive deficit in radiation-induced brain injury model
NSI-189 ameliorates cognitive deficit in radiation-induced brain injury model Spatial Cognitive Impairment Episodic Memory Retention Novel Place Recognition Task Novel Object Recognition Task OPS pc0.05 eo 2 + \ a & | eee earn 2 fol + we : . ‘ShamiRR + vahide Ws veice AR NSBR ‘Sham RR + vehicle IRR+
Slide 26: NSI-189 ameliorates motor deficit in ischemic stroke model
NSI-189 ameliorates motor deficit in ischemic stroke model Mean Neurologic Score 05 Bederson Neurologic Test End of dosing sma Stroke + Vehicle (12 weeks) HB Stroke + NSI-189 (12 weeks) | tC Stroke + Vehicle (24 weeks) =D Stroke + NSI-189 (24 weeks) Historical Control (non-stroke) Baseline Day! Day3
Slide 27: NSI-189 induces synaptic remodeling in ischemic stroke brain
NSI-189 induces synaptic remodeling in ischemic stroke brain A. MAP-2 in Cerebral cortex B. MAP-2 in Hippocampus MAP-2 in Cerebral Cortex * pea.c001. 80 MAP-2 in Hippocampus * peiooos —— *peaarat —_t—, L2Week Control 12
Slide 28: NSI-189 increases motor nerve conduction velocity in type 1 and 2 diabetic models
NSI-189 increases motor nerve conduction velocity in type 1 and 2 diabetic models Type 1 Diabetes Model—Iintervention Study Type 2 Diabetes Model—Prevention Study A ‘Treatment tiated a g 6 a4 ry 3a ; —0- Ct + Vehicle $ 4 PDiabetes + Venice 5 Diabetes +NS1189 o FH 3 H o 0 o2 ee On HH Weeks of Diabete
Slide 29: NSI-189 increases small peripheral nerve fiber density in type 1 diabetic model
NSI-189 increases small peripheral nerve fiber density in type 1 diabetic model Paw Skin NF Pav Shin Dermal Nerves re as 22, 5 oe " S908 % 4 >. a "| “ ° 00 ° Number of PGP 9.5-stained profiles in es & 3 ¢ #« the epidermis (intra-epidermal nerve . Pa : a fibers: IENF) Cornea! Sub Basal Nerve Plexus c
Slide 30: NSI-189 - Summary
NSI-189 - Summary * Orally active neurogenic, synaptogenic, neurotrophic compound for treatment of depression, cognitive impairment, and neurodegeneration — Increases hippocampal neurogenesis, hippocampal volume, synaptogenesis, and synaptic plasticity in normal and/or lesioned brain — Novel, unknow