electrophysiology for robust decision making in CNS drug discovery - - PowerPoint PPT Presentation

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electrophysiology for robust decision making in cns drug
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electrophysiology for robust decision making in CNS drug discovery - - PowerPoint PPT Presentation

Sep 26, 2023 139 likes •445 views

in vitro and in vivo electrophysiology for robust decision making in CNS drug discovery programs Robert E. Petroski, PhD MAY 28, 2020 WELCOME TO TODAYS WEBINAR! Marketing & Communication Manager Neuroservices-Alliance in CNS drug

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in vitro and in vivo electrophysiology for robust decision making in CNS drug discovery programs

MAY 28, 2020

Robert E. Petroski, PhD

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WELCOME TO TODAY’S WEBINAR! Marketing & Communication Manager Neuroservices-Alliance

in vitro and in vivo electrophysiology for robust decision making in CNS drug discovery programs 2 Synaptic transmission and plasticity

3

ADHD behavioral assays

May 28 June 11 June 25

…

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WELCOME TO TODAY’S WEBINAR

  • Ask your questions in the question box
  • Bob will answer your questions during the 10 min

Q&A following his presentation

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NEUROSERVICES-ALLIANCE

Worldwide clients Studies Years of cumulated experience

in vitro data

  • Electrophysiology

Translational data between species

  • Rodent
  • Non Human Primate
  • Human

PhD scientists

in vivo data

  • Electrophysiology
  • Behavior
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MEET TODAY’S PANELIST Scientific Liaison Neuroservices-Alliance

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PATCH CLAMP REVOLUTIONIZED NEUROPHYSIOLOGY

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OUTLINE

Opportunities for CNS Drugs CNS Drug Discovery Workflow Electrophysiology Assays

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V. Pain

I. Neuropathic pain II. Visceral pain

II. Neurodegeneration

I. Alzheimer’s disease II. Parkinson’s disease

  • III. Huntington’s disease
  • IV. Motor neuron diseases

V. Others

  • III. Neurodevelopmental disorders

I. Autism spectrum II. Angelman’s syndrome

  • III. Down syndrome
  • IV. Rett syndrome

V. Others

  • IV. Epilepsy

I. Doose syndrome II. Dravet syndrome

  • III. Lafora disease
  • IV. Landau-Kleffner syndrome

V. Lennox-Gastault syndrome

  • VI. Others

I. Psychiatric disorders

i. Anxiety ii. PTSD iii. Depression iv. Schizophrenia v. Bipolar

THERAPEUTIC OPPORTUNITIES FOR CNS DRUGS

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U N IQU E E X P E R T IS E IN N E U R OP H A R MA C OL OGY

THERAPEUTIC STRATEGIES Small molecules Biologics Gene Therapy

Antisense Oligonucleotide Therapy

Stem Cell Therapy

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IND

Stage gates

GO / NOGO

CNS DRUG DISCOVERY WORKFLOW

IND Pre-clinical development Pre-clinical development Lead optimization Lead optimization Screening Screening Target Research Target Research

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IND Pre-clinical development Lead optimization Screening Target Research

Stage gates

GO / NOGO

CNS DRUG DISCOVERY WORKFLOW

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IND Pre-clinical development Lead optimization Screening Target Research

I. Target validation

  • II. Target signaling pathways
  • III. Target mechanisms
  • IV. Target biomarkers

CNS DRUG DISCOVERY WORKFLOW

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TARGET RESEARCH

field Excitatory Post-Synaptic Potentials (fEPSP)

dentate gyrus CA3 CA1

MEA Hippocampal slice R6/2 (Huntington’s)

10 20 30 40 50 60 70 1.0 1.4 1.8 2.2

R6/2 WT

Time (min) Normalized fEPSP amplitude

HFS

Tg2576 (Alzheimer’s)

10 20 30 40 50 60 70 1.0 1.4 1.8 2.2

Tg2576 WT

Time (min) Normalized fEPSP amplitude

HFS

baseline fEPSPs after TBS
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IND Pre-clinical development Lead optimization Screening Target Research

I. HTS assays

Engineered “kit assays” using surrogate endpoints

CNS DRUG DISCOVERY WORKFLOW

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IND Pre-clinical development Lead optimization Screening Target Research

I. Cellular assays

  • II. Brain slices assays

CNS DRUG DISCOVERY WORKFLOW

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INTRISIC EXCITABILITY OF NEURONS SYNAPTIC TRANSMISSION BETWEEN NEURONS SYNAPTIC PLASTICITY INTRISIC EXCITABILITY OF NETWORKS VOLTAGE-GATED ION CHANNELS LIGAND-GATED ION CHANNELS Field excitatory postsynaptic potentials (fEPSPs) Evoked excitatory postsynaptic currents (EPSCs) Evoked inhibitory postsynaptic currents (IPSCs) Miniature spontaneous excitatory postsynaptic currents (mEPSCs) Miniature spontaneous inhibitory postsynaptic currents (mIPSCs) Spontaneous action potential firing rate resting membrane potential (Vm) Input resistance (Rm) Evoked action potential threshold (rheobase) After hyperpolarizing potential (AHP) Long-term potentiation (LTP) Long-term depression (LTD) Chemical LTD Glutamate receptors: AMPA Glutamate receptors: NMDA GABAA receptors Nicotinic receptors Na currents K currents Ca currents Population spikes (pop spikes) Epileptiform discharges (EDs) Oscillations

SIGNALS / ENDPOINTS

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GABAA CURRENT FROM CULTURED NEURON

recording electrode puffer pipette

250 pA 2 sec 3 uM GABA (100 ms)

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INDIPLON POTENTIATES NATIVE GABAA RECEPTOR CURRENTS

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10 30 50 70 1 2 3 4 5

Time (min) Firing rate (normalized)

1 µM carbachol 20 40 60 80 100 120 140 0.0 0.5 1.0 1.5 2.0 0.1 0.3 0.5 1 3

carbachol (µM) Pirenzepine (100 nM) Pirenzepine (100 nM) + carbachol (µM)

1

Time (min) Firing rate (normalized)

  • 8
  • 7
  • 6
  • 5

0.0 0.5 1.0 1.5 2.0

EC50 = 0.23 µM EC50 = 1.29 µM nH = 1.78

Carbachol Carbachol + 100 nM Pirenzepine

log [Carbachol], M Firing rate (normalised)

MEA RECORDING OF SPONTANEOUS FIRING IN HIPPOCAMPAL SLICES

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PATCH CLAMP FROM PREFRONTAL CORTEX SLICES

50 mV 500 ms PV positive interneuron (layer 2/3) pyramidal neuron (layer V)

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IND Pre-clinical development Lead optimization Screening Target Research

I. In vivo target engagement

  • II. Target engagement

in human slices

CNS DRUG DISCOVERY WORKFLOW

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IN VIVO RECORDING METHODS

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IN VIVO SIGNALS

SINGLE UNITS (HIGHPASS FILTERED >300 HZ) LFPs ARE SLOW (LOWPASS FILTERED <200 Hz)

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LOCAL FIELD POTENTIALS and QUANTITATIVE EEG

LFPs are phasic or oscillatory

  • Phasic signals include evoked potentials,

sharp waves, ictal spikes

  • Oscillatory signals include slow wave sleep,

hippocampal theta, ripples

LFPs are a great marker of large-scale synchronization

  • LFPs in rodents can be a preclinical

biomarker of target engagement

  • LFPs in rodents are a translational

biomarker for clinical EEG

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wake SWS REM

inter-ictal spike rate (Hz)

0.00 0.02 0.04 0.06 0.08

PRE-ICTAL SPIKES IN HIPPOCAMPUS OF FREELY-MOVING Tg2576 MICE

Pre-ictal spikes Pre-ictal spikes increased during REM

VEH LEV inter-ictal firing rate (Hz) 0.000 0.005 0.010 0.015 0.020 VEH LEV inter-ictal firing rate (Hz) 0.000 0.005 0.010 0.015 0.020 VEH LEV inter-ictal firing rate (Hz) 0.00 0.02 0.04 0.06 0.08

WAKE SWS REM

Levetiracetam reduces pre-ictal spikes

p= 0.030 p= 0.069 p= 0.026

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HUMAN BRAIN SLICES

Brain slices

Tissue from brain resection

20 mV 500 ms

Hippocampal pyramidal neuron from human brain slice

50 ms 20 pA Spontaneous EPSCs recorded in voltage clamp Action potential train Recorded in current clamp

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HUMAN IPSC-DERIVED NEURONS

M E A S U R E M E N T S

  • Current clamp (excitability)
  • Voltage clamp (ion channels)
  • Commercial iPSCs
  • Your proprietary iPSCs

i P S C - D E R I V E D N E U R O N S IND Pre-clinical development Lead optimization Screening Target Research

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OUR VALUE PROPOSITION

QUALITY

One-Stop-shop CRO For CNS

ROBUST AND REPRODUCIBLE DATA

SHORT TURNAROUND

LARGE BANDWIDTH AND TOP LEVEL PLATFORMS

INTERACTION

INTERACTIVE AND EXPERT SUPPORT ALL ALONG THE STUDY

FLEXIBILITY

STEP-BY-STEP STUDY PLANS

EXPERTISE

> 500 CUMULATED YEARS OF EXPERIENCE IN NEUROPHARMACOLOGY

INNOVATION

ACTIVE R&D PROGRAMS

We co-design custom solutions with our clients Every scientific question is unique and so is every solution

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ANY QUESTIONS?

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THANK YOU!

www.neuroservices-alliance.com