In Vitro Efficacy Testing Services for Narcolepsy

We provide robust and sensitive in vitro screening and characterization platforms for accelerating the discovery and screening of potential therapies for Narcolepsy. Our services offer comprehensive evaluation of compound efficacy, potency, and mechanism of action in models relevant to sleep-wake regulation and orexin/hypocretin signaling. Key targets include orexin receptors (OX1R and OX2R), monoamine transporters, serotonergic and noradrenergic receptors, and downstream second messenger pathways. We facilitate the study of pathological processes such as orexin deficiency, dysregulation of neurotransmitter signaling, and impaired G-protein coupled receptor (GPCR) function implicated in Narcolepsy.

Our in vitro testing suite encompasses a range of biochemical and cell-based assays designed to assess ligand binding, receptor activation, signal transduction, and neurotransmitter uptake. These methods enable precise quantification of drug-receptor interactions and downstream effects for pharmacological profiling. By leveraging multiple assay technologies, we provide a comprehensive evaluation of candidate compounds targeting the molecular mechanisms underlying Narcolepsy.

Bioluminescence resonance energy transfer (BRET) assay: Measures real-time protein–protein interactions and receptor activation, particularly useful for studying GPCR signaling relevant to orexin receptors.

Chemiluminescent assay: Offers sensitive detection of cellular or biochemical events, such as enzyme activity or second messenger production, to assess compound efficacy.

Displacement of [125I]-orexin A: Evaluates the binding affinity of test compounds for orexin receptors by measuring their ability to displace radiolabeled orexin A.

Displacement of [3H]-8-OH-DPAT: Assesses serotonergic receptor binding by quantifying the displacement of this selective radioligand, relevant for sleep regulation.

Displacement of [3H]-nisoxetine: Used to investigate norepinephrine transporter binding, which can influence wakefulness and alertness.

Displacement of [3H]-paroxetine: Measures compound affinity for the serotonin transporter, important for modulating sleep and mood.

Fluorescent assay: Enables high-throughput detection of cellular events such as receptor activation or second messenger changes using fluorescence-based readouts.

Homogeneous Time Resolved Fluorescence (HTRF) assay: Combines FRET technology with time-resolved detection to quantify protein interactions or second messenger levels in a homogeneous format.

Luciferine/luciferase assay: Quantifies cellular activity or gene expression by detecting bioluminescence, useful for monitoring GPCR activation or downstream signaling.

Radioactivity assay: General category for assays utilizing radiolabeled ligands to quantify binding or functional responses of receptors or transporters.

[35S]-GTPgammaS binding assay: Measures receptor-mediated G-protein activation, providing insights into GPCR signaling efficacy.

cAMP accumulation assay: Quantifies intracellular cAMP levels as a downstream marker of GPCR activation or inhibition, relevant for orexin receptor pharmacology.

We measure a variety of pharmacological parameters to characterize compound potency, efficacy, and binding affinity. These quantitative metrics are essential for comparing candidate molecules and optimizing lead selection during drug development. Accurate determination of these parameters guides decision-making and risk assessment in preclinical research.

EC-50: The concentration of a compound required to achieve half-maximal effect; critical for assessing drug potency in functional assays.

IC-50: The concentration of an inhibitor needed to reduce a specific biological or biochemical function by 50%; important for evaluating antagonist or inhibitory activity.

Ki: The equilibrium dissociation constant for inhibitor binding; used to determine binding affinity and selectivity of a compound for its target.

pEC-50: The negative logarithm of the EC-50 value; provides a more convenient and standardized measure of compound potency for comparison across studies.

Recommended In Vitro Efficacy Tests

5-Hydroxytryptamine Receptor 1A

The 5-Hydroxytryptamine Receptor 1A (5-HT1A) modulates serotonin signaling, influencing sleep-wake cycles and playing a role in narcolepsy pathophysiology. Testing 5-HT1A interactions is crucial for identifying and optimizing narcolepsy therapeutics. Our service employs [3H]-8-OH-DPAT displacement and fluorescent assays to evaluate compound binding and activity. Key parameters measured include EC50 (potency) and Ki (binding affinity), ensuring robust pharmacological profiling for drug development.

Histamine Receptor H3

Our Histamine Receptor H3 testing service supports narcolepsy drug development by targeting H3’s key role in sleep-wake regulation. This testing is crucial for identifying and optimizing compounds that modulate H3 activity. We utilize HTRF, [35S]-GTPγS binding, and luciferin/luciferase assays to assess drug efficacy, with IC50 determination as the main parameter for quantifying compound potency. Accurate H3 profiling accelerates the development of effective narcolepsy therapeutics.

Hypocretin Receptor 1

The Hypocretin Receptor 1 (HCRTR1) is crucial in regulating wakefulness, and its dysfunction is linked to narcolepsy. Our testing service supports drug development by assessing compounds’ activity at HCRTR1 using fluorescent assays and [125I]-orexin A displacement. Key pharmacological parameters, including EC-50, pEC-50, and IC-50, are determined, enabling precise evaluation of drug potency and efficacy for narcolepsy therapeutics.

Solute Carrier Family 6 Member 2

The Solute Carrier Family 6 Member 2 (SLC6A2) encodes the norepinephrine transporter, implicated in narcolepsy pathophysiology. Testing SLC6A2 activity is crucial for developing effective narcolepsy therapeutics. Our assay utilizes [3H]-nisoxetine displacement and radioactivity measurement to evaluate compound interactions. Key parameters, IC50 and Ki, provide quantitative insights into inhibitor potency and binding affinity, supporting precise drug candidate optimization.

Solute Carrier Family 6 Member 3

Solute Carrier Family 6 Member 3 (SLC6A3) encodes the dopamine transporter, crucial in regulating dopamine levels linked to Narcolepsy. Testing SLC6A3 function is vital for developing targeted Narcolepsy therapies. Our service offers both fluorescent and radioactivity assays to assess compound effects on transporter activity, with IC-50 determination as the primary parameter to quantify inhibitory potency. This enables efficient screening and optimization of drug candidates.

Solute Carrier Family 6 Member 4

The Solute Carrier Family 6 Member 4 (SLC6A4/serotonin transporter) is implicated in Narcolepsy pathophysiology by modulating serotonin reuptake. Testing its activity is essential for identifying and optimizing therapeutics targeting this transporter. Our service utilizes fluorescent and radioactivity assays, including [3H]-paroxetine displacement, to assess compound efficacy. Key parameters measured are IC-50 and Ki, enabling precise evaluation of drug potency and binding affinity for Narcolepsy drug development.

Trace Amine Associated Receptor 1

Trace Amine Associated Receptor 1 (TAAR1) is implicated in sleep-wake regulation and is a promising target for Narcolepsy drug development. Our TAAR1 testing service employs advanced BRET, chemiluminescent, and cAMP accumulation assays to assess ligand activity and receptor signaling. Key pharmacological parameters measured include EC-50 and pEC-50, enabling precise evaluation of compound potency and efficacy for targeted therapeutics in Narcolepsy.