In Vitro Efficacy Testing Services for Pulmonary Arterial Hypertension

We provide robust and sensitive in vitro screening and characterization platforms for accelerating the discovery and screening of potential therapies for Pulmonary Arterial Hypertension (PAH). Our services enable comprehensive evaluation of candidate compounds, focusing on their effects on vascular smooth muscle cell proliferation, endothelial dysfunction, and signaling pathways implicated in PAH. Key targets include endothelin receptors, prostacyclin pathways, phosphodiesterases (notably PDE5), and cGMP-mediated signaling. We assess critical pathological processes such as vasoconstriction, cellular remodeling, and inflammation associated with the progression of PAH.

Our in vitro testing methods encompass a wide range of biochemical and cell-based assays designed to measure compound efficacy, mechanism of action, and pathway modulation. These assays utilize luminescent, fluorescent, and radiometric technologies to ensure high sensitivity, specificity, and quantitative analysis of drug-target interactions or functional outcomes. The overall goal is to generate reliable data to guide lead optimization and therapeutic development.

ATP assay: Quantifies cellular ATP levels as an indicator of cell viability, metabolic activity, and cytotoxicity, relevant for assessing compound effects on pulmonary vascular cells.

Bioluminescence resonance energy transfer (BRET) assay: Detects real-time protein-protein interactions or signaling events in live cells, useful for monitoring receptor activation and pathway modulation in PAH.

Chemiluminescent assay: Measures specific biomolecules or enzymatic activities using luminescent readouts, providing sensitive detection of target engagement or downstream signaling.

ELISA assay: Enables quantitative measurement of proteins, peptides, or cytokines, such as endothelin-1 or inflammatory mediators, to evaluate pathway-specific effects.

Fluorescence resonance energy transfer (FRET) assay: Monitors molecular interactions or conformational changes, supporting mechanistic studies of signaling cascades relevant to PAH.

Fluorescent assay: Offers versatile detection of cellular or biochemical events, including calcium flux, membrane potential, or reactive oxygen species, important in PAH pathology.

Fluorescent polarization assay: Measures binding interactions between small molecules and target proteins, facilitating high-throughput affinity screening.

Homogeneous Time Resolved Fluorescence (HTRF) assay: Combines FRET with time-resolved measurements for sensitive, homogeneous detection of signaling molecules or second messengers like cGMP.

RNA assay: Quantifies gene expression changes for relevant markers or targets, enabling assessment of compound impact on transcriptional regulation.

Radioactivity assay: Employs radiolabeled substrates or ligands to measure enzyme activities or receptor binding with high sensitivity.

cGMP as substrate: Utilizes cGMP quantification to assess modulation of the nitric oxide signaling pathway, a key therapeutic target in PAH.

Our assays provide a range of key pharmacological parameters critical for drug development and candidate prioritization. These parameters include potency, efficacy, and selectivity metrics that inform structure-activity relationships and therapeutic potential. Accurate measurement of these values enables data-driven decision-making throughout the lead optimization process.

EC-50: The concentration of a compound that produces 50% of its maximal effect, representing compound potency and informing dose selection.

IC-50: The concentration of inhibitor required to reduce a specific biological activity by 50%, used to compare compound inhibitory strength on target enzymes or pathways.

MEC (Minimum Effective Concentration): The lowest concentration at which a compound demonstrates a measurable biological effect, guiding therapeutic window estimation.

pIC-50: The negative logarithm of the IC-50 value, providing a standardized comparison of inhibitory potency across compounds and assays.

Recommended In Vitro Efficacy Tests

Abl Proto-Oncogene 1, Non-Receptor Tyrosine Kinase

Abl Proto-Oncogene 1, Non-Receptor Tyrosine Kinase (ABL1) regulates vascular remodeling and smooth muscle proliferation in Pulmonary Arterial Hypertension (PAH). Testing ABL1 activity is crucial for assessing drug efficacy and safety in PAH drug development. Key methods include kinase activity assays, Western blotting, and qPCR. Main parameters assessed are ABL1 phosphorylation status, expression levels, and downstream signaling pathway activation.

Colony Stimulating Factor 1 Receptor

Colony Stimulating Factor 1 Receptor (CSF1R) is implicated in vascular remodeling and inflammation in Pulmonary Arterial Hypertension (PAH). Testing CSF1R inhibition is crucial for identifying promising drug candidates targeting these pathways. Our service utilizes Bioluminescence Resonance Energy Transfer (BRET) and ATP assays to assess compound activity, providing precise IC-50 values to quantify inhibitory potency, thereby accelerating the development of effective PAH therapies.

Endothelin Receptor Type A

Endothelin Receptor Type A (ETA) mediates vasoconstriction and proliferation in Pulmonary Arterial Hypertension (PAH). ETA testing is crucial for evaluating drug efficacy and selectivity in PAH therapy development. Key methods include radioligand binding assays and functional cell-based assays. Main parameters assessed are binding affinity (Ki), receptor selectivity, and functional inhibition (IC50), enabling optimization of candidate drugs targeting the ETA pathway in PAH.

Kit Proto-Oncogene, Receptor Tyrosine Kinase

Kit Proto-Oncogene, Receptor Tyrosine Kinase is implicated in abnormal cell proliferation in Pulmonary Arterial Hypertension (PAH). Testing its activity is crucial for evaluating targeted therapies in PAH drug development. Using a chemiluminescent assay, this service quantitatively measures kinase inhibition, with IC-50 as the main parameter to determine compound potency. This enables efficient screening of drug candidates targeting the Kit pathway.

Nfe2 Like Bzip Transcription Factor 2

The Nfe2 Like Bzip Transcription Factor 2 (Nrf2) plays a pivotal role in oxidative stress response implicated in Pulmonary Arterial Hypertension (PAH) pathogenesis. Testing its activity is crucial for evaluating candidate drugs targeting this pathway. Our service employs sensitive chemiluminescent assays to quantify Nrf2 modulation, providing key parameters such as EC-50 and MEC, essential for determining compound potency and minimum effective concentration in PAH drug development.

Peroxisome Proliferator Activated Receptor Gamma

Peroxisome Proliferator Activated Receptor Gamma (PPARγ) regulates vascular remodeling and inflammation in Pulmonary Arterial Hypertension (PAH). Testing PPARγ activity is crucial for evaluating novel PAH therapeutics targeting this pathway. Our service utilizes RNA assays to quantify PPARγ-related gene expression, providing the Minimum Effective Concentration (MEC) of candidate drugs, ensuring precise assessment of therapeutic efficacy in PAH drug development.

Phosphodiesterase 5A

Phosphodiesterase 5A (PDE5A) regulates cGMP levels, influencing vascular tone and contributing to Pulmonary Arterial Hypertension (PAH). PDE5A testing is vital for evaluating drug efficacy in PAH development. Our service employs fluorescent assays, fluorescent polarization assays, and radioactivity assays using cGMP as a substrate. Key parameters measured include IC50 and pIC50, enabling precise assessment of compound potency and inhibitor effectiveness in PDE5A-targeted drug discovery.

Platelet Derived Growth Factor Receptor Alpha

Platelet Derived Growth Factor Receptor Alpha (PDGFRα) promotes vascular remodeling in Pulmonary Arterial Hypertension (PAH). Testing PDGFRα is vital for identifying therapeutic targets and monitoring drug efficacy. Key methods include immunohistochemistry, Western blotting, and qPCR. Main parameters assessed are PDGFRα expression levels, phosphorylation status, and downstream signaling activity, enabling precise evaluation of drug effects in PAH research and development.

Platelet Derived Growth Factor Receptor Beta

Platelet Derived Growth Factor Receptor Beta (PDGFRβ) plays a critical role in vascular remodeling and smooth muscle cell proliferation in Pulmonary Arterial Hypertension (PAH). Testing PDGFRβ activity is essential for evaluating drug candidates targeting this pathway. Our service employs FRET, HTRF, and ELISA assays to assess compound effects on PDGFRβ, with IC-50 as the primary parameter, enabling precise measurement of inhibitor potency for PAH drug development.

Prostaglandin I2 Receptor

Prostaglandin I2 Receptor (IP receptor) plays a key role in vasodilation and inhibition of proliferation in Pulmonary Arterial Hypertension (PAH). Accurate testing of IP receptor modulation is crucial for developing effective PAH drugs. Our service utilizes ELISA and Homogeneous Time Resolved Fluorescence (HTRF) assays to quantify receptor activity. Main parameters measured include EC50 and IC50, providing essential data for evaluating drug potency and efficacy.