In Vitro Efficacy Testing Services for Obesity

We provide robust and sensitive in vitro screening and characterization platforms for accelerating the discovery and screening of potential therapies for obesity. Our services offer comprehensive analysis of compound efficacy, binding, and signaling relevant to obesity-associated targets such as glucagon-like peptide-1 (GLP-1) receptor, gastric inhibitory polypeptide (GIP) receptor, and other metabolic regulators. The key targets and pathways include those involved in energy balance, appetite regulation, insulin sensitivity, and adipogenesis. We assess the modulation of these pathways and proteins to address underlying pathological processes such as dysregulated glucose metabolism, impaired satiety signaling, and abnormal lipid storage.

Our in vitro testing services encompass a diverse suite of assay formats, including biochemical, biophysical, and cell-based platforms to evaluate compound-target interactions, functional responses, and signaling events. These assays enable detailed characterization of drug candidates' mechanism of action, potency, and selectivity. The methods are designed to provide critical insights for early-stage screening and lead optimization in obesity drug discovery.

Arrestin protease recruitment assay: Evaluates compound-induced recruitment of arrestin to target receptors, indicative of receptor activation and signaling bias relevant to metabolic modulation.

Bioluminescent assay: Utilizes light-emitting reactions to quantify cellular or enzymatic responses, enabling sensitive detection of target engagement or biological activity.

Chemiluminescent assay: Measures light emission from chemical reactions to analyze binding or activity, offering high sensitivity for detecting low-abundance targets.

Chemiluminescent assay (with human serum albumin): Incorporates human serum albumin to mimic physiological conditions and assess compound activity in a more relevant matrix.

Competitive binding assay (with 0.005% human serum albumin): Determines the binding affinity of test compounds by competing with labeled ligands, using low serum albumin for physiological relevance.

Competitive binding assay (with 2% human serum albumin): Similar to the above, with higher serum albumin to further emulate in vivo conditions and assess potential protein binding effects.

Displacement of [125I]-gastric inhibitory polypeptide: Measures the ability of compounds to displace radiolabeled GIP, indicating binding affinity and potential receptor modulation.

Displacement of [125I]-glucagon: Assesses compound binding to the glucagon receptor by displacement of radiolabeled glucagon, relevant for metabolic regulation studies.

Displacement of [125I]-glucagon-like polypeptide 1: Evaluates binding to the GLP-1 receptor, a key target in obesity therapy, by radioligand displacement.

ELISA assay: Quantifies proteins, peptides, or signaling molecules using enzyme-linked detection, supporting measurement of biomarkers and cytokines related to obesity.

Fluorescence resonance energy transfer (FRET) assay: Detects changes in proximity between fluorophores to study molecular interactions and receptor activation.

Fluorescence resonance energy transfer (FRET) assay (with 2% human serum albumin): Adapts FRET to a serum albumin-containing environment, enhancing physiological relevance.

Fluorescent assay: Uses fluorescent signals to assess enzyme activity, cellular responses, or protein interactions in obesity-related pathways.

Fluorescent polarization assay: Measures changes in fluorescence polarization to evaluate binding interactions or conformational changes in target proteins.

Homogeneous Time Resolved Fluorescence (HTRF) assay: Employs time-resolved fluorescence for sensitive detection of molecular interactions and signaling events.

Homogeneous Time Resolved Fluorescence (HTRF) assay (2% human serum albumin): Conducts HTRF in the presence of serum albumin to approximate in vivo binding conditions.

Homogeneous Time Resolved Fluorescence (HTRF) assay (with fetal bovine serum/bovine serum albumin): Uses animal serum proteins to further simulate physiological assay environments.

Luciferine/luciferase assay: Detects luciferase enzyme activity, commonly used to quantify gene expression and signaling in metabolic pathways.

Luciferine/luciferase assay (with 0.005% human serum albumin): Incorporates low serum albumin to replicate plasma protein effects on compound activity.

Luciferine/luciferase assay (with 2% human serum albumin): Uses higher albumin concentrations for enhanced physiological relevance in activity measurements.

Plasma, human: Utilizes human plasma as an assay matrix to evaluate compound stability, protein binding, or activity in a native biological context.

RNA assay: Quantifies gene expression changes in response to treatment, supporting studies of transcriptional regulation in obesity.

Radioactivity assay: Uses radiolabeled compounds or ligands to assess binding, uptake, or metabolic processing with high sensitivity.

Scintillation proximity assay (SPA): Measures radioligand binding without separation steps, allowing high-throughput detection of molecular interactions.

Surface plasmon resonance assay: Real-time, label-free analysis of binding kinetics and affinity between compounds and targets.

With 0.5% human plasma: Conducts assays with added human plasma to assess effects of plasma proteins on compound activity.

With human serum albumin (1%): Assays incorporating 1% albumin to study protein binding and bioavailability aspects.

cAMP accumulation assay: Measures intracellular cAMP levels to monitor GPCR activation, a key signaling pathway in metabolic regulation.

We measure a range of pharmacological parameters that capture the potency, efficacy, and binding characteristics of test compounds. These parameters, including EC-50, IC-50, Kd, and Ki, are essential for determining the functional and biophysical profiles of drug candidates targeting obesity. Accurate quantification of these metrics supports informed decision-making during hit validation, lead optimization, and candidate selection.

EC-50: The concentration of a compound that produces 50% of its maximal effect, reflecting compound potency in functional assays.

IC-50: The concentration required to inhibit a biological or biochemical function by 50%, widely used to assess antagonist or inhibitor efficacy.

Kd: The equilibrium dissociation constant, indicating the affinity between a ligand and its target; lower Kd values denote higher affinity.

Ki: The inhibition constant, representing the potency of an inhibitor in binding assays, important for characterizing competitive interactions.

MEC: Minimum effective concentration, the lowest concentration at which a compound elicits a measurable effect, useful for dose selection.

MIC: Minimum inhibitory concentration, the lowest concentration needed to inhibit a specific biological process, aiding in efficacy evaluation.

pEC-50: The negative logarithm of the EC-50, providing a more convenient scale to compare compound potencies.

pIC-50: The negative logarithm of the IC-50, enabling straightforward comparison of inhibitor strengths across compounds.

Recommended In Vitro Efficacy Tests

5-Hydroxytryptamine Receptor 2C

The 5-Hydroxytryptamine Receptor 2C (5-HT2C) regulates appetite and energy balance, making it a crucial target in obesity drug development. Testing 5-HT2C activity helps identify compounds that modulate this receptor, aiding anti-obesity therapy design. Our service uses FRET, luciferin/luciferase, and fluorescent assays to assess ligand interactions. Key parameters measured include pEC-50, MEC, EC-50, and Ki, enabling precise evaluation of compound potency and efficacy.

Carbonic Anhydrase 2

Carbonic Anhydrase 2 (CA2) is implicated in adipocyte metabolism and energy balance, making it a potential target in obesity drug development. Testing CA2 activity is crucial for evaluating drug efficacy and off-target effects. Key methods include enzyme activity assays, ELISA, and Western blotting. Main parameters assessed are CA2 expression levels, enzymatic activity, and inhibitor potency, providing essential insights into compound mechanisms and therapeutic potential.

Epidermal Growth Factor Receptor

Epidermal Growth Factor Receptor (EGFR) plays a significant role in obesity by regulating adipocyte differentiation and metabolism. EGFR testing is crucial in obesity drug development to identify therapeutic targets and assess drug efficacy. Our service utilizes sensitive RNA assays to quantify EGFR expression levels, providing insights into molecular mechanisms. Key parameters, such as the minimum inhibitory concentration (MIC), are measured to evaluate drug potency against EGFR-mediated pathways, supporting efficient candidate selection.

Fatty Acid Synthase

Fatty Acid Synthase (FAS) is crucial in lipid biosynthesis and is often upregulated in obesity, making it a valuable target for drug development. FAS testing evaluates compound efficacy using RNA and radioactivity assays, which measure expression and enzymatic activity, respectively. The primary parameter assessed is IC-50, indicating the concentration required to inhibit FAS activity by 50%, essential for identifying potent anti-obesity candidates.

Gastric Inhibitory Polypeptide Receptor

The Gastric Inhibitory Polypeptide Receptor (GIPR) regulates insulin secretion and fat metabolism, making it a key target in obesity drug development. GIPR testing is crucial for evaluating candidate compounds’ efficacy and selectivity. Our service includes [125I]-GIP displacement, cAMP accumulation, luciferase, HTRF, arrestin recruitment, FRET, and bioluminescent/fluorescent assays. We provide critical pharmacological parameters, including pEC-50, EC-50, and Ki, to support informed drug candidate selection.

Glucagon Like Peptide 1 Receptor

Glucagon Like Peptide 1 Receptor (GLP-1R) plays a pivotal role in obesity by regulating appetite and glucose metabolism. Accurate GLP-1R testing is vital for obesity drug development to assess candidate drug efficacy and receptor selectivity. Our service utilizes advanced methods—such as luciferase/luciferin, HTRF, chemiluminescence, FRET, ELISA, and binding assays—in physiological matrices. Key pharmacological parameters measured include pEC-50, EC-50, IC-50, pIC-50, Kd, and Ki, enabling robust candidate evaluation.

Glucagon Receptor

The Glucagon Receptor plays a crucial role in glucose and energy metabolism, making it a key target in obesity drug development. Glucagon Receptor testing enables precise evaluation of drug candidates’ efficacy and selectivity using advanced methods such as cAMP accumulation, luciferase, [125I]-glucagon displacement, HTRF, FRET, chemiluminescence, and fluorescent assays in relevant biological matrices. Key parameters measured include pEC-50, IC-50, EC-50, and Ki, supporting data-driven therapeutic development.

Hypocretin Receptor 2

Our Hypocretin Receptor 2 (HCRTR2) testing service supports obesity drug development by evaluating compounds that modulate this receptor, which regulates appetite and energy balance. Utilizing advanced fluorescent and Homogeneous Time Resolved Fluorescence (HTRF) assays, we determine key pharmacological parameters, including IC-50 and EC-50 values, enabling precise assessment of compound potency and efficacy for targeted anti-obesity therapies.

Opioid Receptor Kappa 1

Opioid Receptor Kappa 1 (OPRK1) regulates appetite and energy balance, making it a significant target in obesity drug development. Our testing service utilizes the arrestin protease recruitment assay to evaluate compound interactions with OPRK1. Key parameters measured include IC-50 and EC-50, providing critical data on compound potency and efficacy. This service supports the identification and optimization of effective anti-obesity therapeutics.

Opioid Receptor Mu 1

Opioid Receptor Mu 1 (MOR-1) is implicated in regulating appetite and energy balance, making it a key target in obesity drug development. Our testing service evaluates drug candidates’ activity on MOR-1 using the arrestin protease recruitment assay. Critical parameters measured include IC-50 and EC-50 values, enabling precise assessment of compound potency and efficacy, which is essential for advancing effective anti-obesity therapeutics.

Pancreatic Lipase

Pancreatic Lipase is crucial for dietary fat digestion, and its inhibition is a proven strategy in obesity drug development. Pancreatic Lipase testing evaluates a compound’s ability to block this enzyme, helping identify potential anti-obesity agents. Using ELISA assays, we measure enzyme activity and determine the IC-50, the concentration at which an inhibitor reduces enzyme activity by 50%, providing essential data for drug efficacy assessment.

Solute Carrier Family 6 Member 4

Solute Carrier Family 6 Member 4 (SLC6A4) regulates serotonin reuptake, influencing appetite and obesity risk. Testing SLC6A4 is crucial for developing anti-obesity drugs targeting this pathway. Using radioactivity assays and scintillation proximity assays (SPA), we determine key parameters such as IC50 and Ki to assess compound potency and binding affinity, enabling effective candidate selection and optimization in obesity drug development.