In Vitro Efficacy Testing Services for Interstitial Cystitis

We provide robust and sensitive in vitro screening and characterization platforms for accelerating the discovery and screening of potential therapies for interstitial cystitis. Our service offers comprehensive efficacy testing using cellular, molecular, and binding assays to assess drug candidates targeting key aspects of interstitial cystitis pathology. Relevant targets include urothelial receptors, inflammatory mediators, pain pathway modulators, and ion channels implicated in bladder dysfunction. We can evaluate drug effects on inflammation, neurotransmitter activity, barrier function, and nociceptive signaling associated with interstitial cystitis.

Our in vitro testing methods encompass a diverse array of biochemical, biophysical, and cell-based assays designed to elucidate efficacy, mechanism of action, and interaction profiles of candidate compounds. We utilize advanced technologies such as radioligand binding, fluorescence-based assays, electrophysiological recordings, and immunoassays to provide comprehensive pharmacological characterization. These approaches allow for precise evaluation of drug-target engagement, functional activity, and cellular responses.

ATP assay: Assesses cellular viability and metabolic activity by quantifying ATP levels, useful for evaluating cytotoxicity and cell health in response to therapies.

Biolayer interferometry assay: Measures real-time biomolecular interactions, enabling kinetic analysis of drug binding to disease-relevant targets.

Calcein deposition assay: Uses calcein dye to detect cell membrane integrity and permeability, relevant for assessing urothelial barrier function.

Competitive binding assay: Determines the affinity of compounds for specific receptors or transporters by measuring displacement of known ligands.

Displacement of [3H]-citalopram: Evaluates binding to serotonin transporters, important for modulating pain and inflammation.

Displacement of [3H]-imipramine: Assesses compound interaction with monoamine transporters, relevant to pain transmission.

Displacement of [3H]-nicotinic acid: Measures binding to specific receptors involved in inflammation and metabolic pathways.

Displacement of [3H]-nisoxetine: Tests affinity for norepinephrine transporters, linked to pain signaling modulation.

Displacement of [3H]-nociceptin: Assesses binding to nociceptin receptors, involved in pain and bladder function.

Displacement of [3H]-paroxetine: Evaluates interaction with serotonin transporters, central to neuromodulatory pathways.

ELISA assay: Quantifies proteins, cytokines, or biomarkers to monitor inflammatory responses or therapeutic effects.

Endothelial cells (umbilical vein), human: Utilizes primary human endothelial cells to test vascular and inflammatory responses to candidate drugs.

Fluorescent (Aequorin) assay: Measures changes in intracellular calcium, reflecting receptor activation or inhibition.

Fluorescent assay: Broadly detects molecular or cellular changes using fluorescence, adaptable to diverse targets.

Patch-clamp assay: Records ionic currents across cell membranes to study drug effects on ion channels critical in bladder function.

Patch-clamp assay (-60 mV): Specialized patch-clamp at -60 mV to assess voltage-dependent ion channel activity.

RNA assay: Measures gene expression changes, providing insights into molecular mechanisms and target engagement.

Radioactivity assay: Quantifies ligand binding or enzymatic activity using radiolabeled compounds for high sensitivity.

Surface plasmon resonance assay: Provides label-free, real-time analysis of biomolecular interactions and binding kinetics.

Voltage-clamp assay (-60 mV): Assesses ion channel function by controlling membrane potential, vital for evaluating neuromodulatory effects.

Whole cell recording: Captures electrical activity of entire cells to assess functional responses to treatments.

Whole-cell patch-clamp assay (-80 mV): Measures drug effects on ion channel activity at hyperpolarized potentials, relevant for neuronal signaling.

[35S]-GTPgammaS binding assay: Evaluates G-protein coupled receptor activation, important for signaling pathway analysis.

cAMP accumulation assay: Quantifies intracellular cAMP levels to assess receptor-mediated signaling relevant to inflammation and cell signaling.

Our assays yield a range of quantitative pharmacological parameters including potency, efficacy, and binding affinity, which are central to drug candidate evaluation. These parameters, such as EC-50, IC-50, and Ki, provide critical insights into the strength, selectivity, and functional effects of compounds on target proteins. Accurate measurement of these values guides optimization and selection of promising therapeutic leads.

EC-50: The concentration of a compound that produces 50% of its maximal effect; indicates functional potency.

ED-50: The dose at which 50% of the maximal therapeutic effect is observed; used to compare effective dosages.

IC-50: The concentration at which a compound inhibits a specific biological or biochemical function by 50%; measures inhibitory potency.

Kd: The equilibrium dissociation constant, reflecting the affinity between a drug and its target; lower Kd indicates higher affinity.

Ki: The inhibition constant, quantifying the binding affinity of an inhibitor for its target; essential for characterizing competitive binding.

Ki(h): Human-specific inhibition constant, providing affinity data for compounds in human systems.

MED: Minimum effective dose, representing the lowest dose at which a biological effect is observed; important for dosing strategies.

MIC: Minimum inhibitory concentration, the lowest concentration required to inhibit microbial growth; relevant for anti-infective agents.

pIC-50: The negative logarithm of IC-50, used for convenient comparison of inhibitory potencies.

pKi: The negative logarithm of Ki, allowing for standardized reporting and comparison of binding affinities.

Recommended In Vitro Efficacy Tests

Adrenoceptor Beta 3

The Adrenoceptor Beta 3 testing service evaluates the receptor’s role in modulating bladder function, crucial for interstitial cystitis drug development. This service uses cAMP accumulation assays to assess compound activity, determining Minimum Effective Dose (MED), half-maximal inhibitory concentration (IC-50), and half-maximal effective concentration (EC-50). Accurate profiling of Beta 3 agonists or antagonists aids in identifying promising therapeutic candidates for interstitial cystitis.

Atp Binding Cassette Subfamily B Member 1

Atp Binding Cassette Subfamily B Member 1 (ABCB1) modulates drug efflux and impacts therapeutic response in interstitial cystitis. Testing ABCB1 function is crucial for optimizing drug efficacy and minimizing resistance. Our service employs ATP assays, calcein deposition, and fluorescent assays to assess ABCB1 activity. Key parameters measured include Ki(h) for inhibitor potency and MIC for minimal inhibitory concentration, ensuring precise evaluation of candidate drug interactions with ABCB1.

Fibroblast Growth Factor 2

Fibroblast Growth Factor 2 (FGF2) is implicated in the pathophysiology of interstitial cystitis by modulating inflammation and tissue repair. Accurate FGF2 testing is essential for drug development, enabling assessment of therapeutic efficacy. Our service utilizes RNA assays to quantify FGF2 expression, with Minimum Inhibitory Concentration (MIC) as a key parameter, ensuring precise evaluation of drug candidates targeting FGF2-mediated pathways in interstitial cystitis.

Hydroxycarboxylic Acid Receptor 2

Hydroxycarboxylic Acid Receptor 2 (HCA2) modulates inflammatory responses implicated in interstitial cystitis. Testing HCA2 activity is crucial for identifying potential therapeutics targeting this pathway. Our service employs fluorescent (Aequorin) and cAMP accumulation assays, along with [3H]-nicotinic acid displacement, to assess ligand-receptor interactions. Key pharmacological parameters measured include Ki, IC50, and EC50, enabling robust evaluation of compound potency and efficacy for drug development.

Opioid Related Nociceptin Receptor 1

The Opioid Related Nociceptin Receptor 1 (NOP) is implicated in pain modulation and bladder dysfunction in interstitial cystitis. Testing NOP is crucial for developing targeted therapies. Our service assesses drug-receptor interactions via [3H]-nociceptin displacement and [35S]-GTPγS binding assays. Key parameters measured include Ki (binding affinity) and EC50 (functional potency), providing essential data for evaluating candidate drug efficacy and selectivity.

Purinergic Receptor P2X 2

Our Purinergic Receptor P2X2 testing service supports interstitial cystitis drug development by evaluating compounds that modulate P2X2, a receptor implicated in bladder pain and inflammation. Testing is vital for identifying agents that target this pathway. Using whole cell recording, we measure receptor activity and determine compound potency via pIC50 values, offering precise insights into efficacy for therapeutic advancement.

Purinergic Receptor P2X 3

Our Purinergic Receptor P2X3 testing service supports interstitial cystitis drug development by targeting the P2X3 receptor, a key mediator of pain and urinary symptoms. Accurate assessment of P2X3 antagonists is critical for therapeutic advancement. We utilize fluorescent, patch-clamp, voltage-clamp (-60 mV), and whole-cell patch-clamp (-80 mV) assays to evaluate compound efficacy, focusing on IC50 determination as the main pharmacological parameter.

Solute Carrier Family 6 Member 2

The Solute Carrier Family 6 Member 2 (SLC6A2) transporter regulates norepinephrine uptake, implicated in interstitial cystitis pathophysiology. Testing SLC6A2 activity is crucial for identifying potential therapeutic agents. Using [3H]-nisoxetine displacement and radioactivity assays, this service determines compound affinity and efficacy, providing key pharmacological parameters: Ki (inhibitor binding affinity), ED-50 (effective dose), and IC-50 (half-maximal inhibitory concentration), essential for drug candidate evaluation.

Solute Carrier Family 6 Member 4

The Solute Carrier Family 6 Member 4 (SLC6A4) is implicated in interstitial cystitis via serotonin reuptake modulation, influencing bladder pain and inflammation. Testing SLC6A4 interactions is vital for developing effective therapeutics. Our service uses [3H]-imipramine, [3H]-paroxetine, and [3H]-citalopram displacement assays, with radioactivity-based detection, to determine compound affinities. Key parameters measured include Ki, ED-50, pKi, and IC-50, enabling robust evaluation of candidate drug efficacy and potency.

Tumor Necrosis Factor

Tumor Necrosis Factor (TNF) is implicated in the inflammatory processes of interstitial cystitis, making it a key target in drug development. Our TNF testing service employs biolayer interferometry, competitive binding, ELISA, surface plasmon resonance assays, and human umbilical vein endothelial cells to assess drug efficacy. We provide critical parameters, including IC-50, MED, Kd, and MIC, enabling precise evaluation of candidate therapeutics for TNF modulation in interstitial cystitis.