BIBP 3226 Trifluoroacetate: Precision in NPY/NPFF System Research

Overview: Unveiling the Power of a Non-Peptide NPY Y1/NPFF Receptor Antagonist

Understanding the intricate signaling crosstalk between neuropeptide Y (NPY) and neuropeptide FF (NPFF) systems is crucial for unraveling mechanisms underlying anxiety, pain modulation, and cardiovascular regulation. BIBP 3226 trifluoroacetate (CAS: 1068148-47-9) stands out as a highly selective, non-peptide NPY Y1 receptor antagonist and potent NPFF receptor antagonist. With exceptional binding affinities—Ki of 1.1 nM for rat NPY Y1, 79 nM for human NPFF2, and 108 nM for rat NPFF receptors—this compound enables researchers to interrogate the neuropeptide Y and FF receptor pathways with clarity and control.

Mechanistically, BIBP 3226 trifluoroacetate competes with endogenous NPFF, effectively blocking NPFF-induced inhibition of forskolin-stimulated cAMP production. This cAMP signaling inhibition is central for studying downstream physiological roles, particularly in cardiac, neural, and adipose tissue interactions.

Recent advances underscore its importance: Fan et al. (2024) demonstrated that the leptin–NPY/Y1R axis orchestrates arrhythmogenic signaling in epicardial adipose tissue-related cardiac arrhythmias, highlighting the translational value of specific NPY Y1 antagonists in cardiovascular research.

Step-by-Step: Integrating BIBP 3226 Trifluoroacetate into Experimental Workflows

1. Reagent Preparation

• Solubilization: Dissolve BIBP 3226 trifluoroacetate at ≥78 mg/mL in DMSO, ≥73.2 mg/mL in ethanol, or ≥12.13 mg/mL in water (ultrasonic assistance recommended for water).
• Aliquoting: Prepare small aliquots to minimize freeze-thaw cycles; store at -20°C.
• Solution Stability: Use freshly prepared solutions for maximal activity. Avoid long-term storage of working solutions.

2. Cellular Assay Integration

• Coculture Models: BIBP 3226 trifluoroacetate is ideal for advanced coculture systems, such as those combining sympathetic neurons, adipocytes, and cardiomyocytes to recapitulate the adipose-neural axis, as established in Fan et al. (2024).
• Dosing: Typical working concentrations range from 1–100 nM for receptor antagonism, with titrations advised based on cell model sensitivity and desired blockade depth.
• Controls: Always include vehicle (e.g., DMSO) and positive controls (e.g., known Y1 or NPFF agonists/antagonists) to confirm specificity.

3. Functional Readouts

• cAMP Assays: Quantify cAMP levels post-forskolin stimulation to verify NPFF-dependent signaling inhibition.
• Electrophysiology: Monitor arrhythmogenic events in cardiomyocytes to assess the functional impact of Y1/NPFF blockade.
• Immunocytochemistry/Western Blot: Probe for downstream effectors such as NCX and CaMKII to connect receptor antagonism to cellular responses.

Advanced Applications and Comparative Advantages

Dissecting the Adipose-Neural Axis in Cardiac Arrhythmia

The study by Fan et al. (2024) revealed that adipocyte-derived leptin heightens NPY release from sympathetic neurons, activating Y1R on cardiomyocytes and triggering arrhythmias via NCX and CaMKII upregulation. By incorporating BIBP 3226 trifluoroacetate, researchers successfully interrupted this pathogenic signaling cascade, providing a powerful model for cardiovascular regulation research and the development of targeted interventions for atrial fibrillation and related disorders.

Enabling High-Resolution NPY/NPFF System Research

Unlike peptide-based antagonists, this robust non-peptide molecule offers:

• Superior Stability: Off-white solid form, high aqueous and organic solubility, and minimal batch-to-batch variation (purity >98% by HPLC, MS, NMR).
• Versatility: Compatible with in vitro, ex vivo, and in vivo rodent models.
• Specificity: Selectively targets NPY Y1 and NPFF receptors without off-target peptide degradation concerns.
• Translational Relevance: Facilitates mechanistic studies in anxiety research, analgesia mechanism study, and metabolic-cardiovascular cross talk.

Complementary Literature and Extended Insights

In "BIBP 3226 Trifluoroacetate: Unraveling the NPY/NPFF Axis", the compound’s role as a non-peptide NPY Y1 receptor antagonist is positioned as a pivotal tool for probing cardiac and neural signaling. This work complements Fan et al. by offering a mechanistic deep dive into receptor pathway modulation. Meanwhile, "Precision Tool for NPY/NPFF System Studies" expands on BIBP 3226’s compatibility with complex coculture models, mirroring the advanced arrhythmia model used by Fan et al., and underscores its practical edge in translational settings. Together, these resources illustrate how BIBP 3226 trifluoroacetate’s versatility and reliability extend across experimental paradigms, from molecular assays to multicellular systems.

Troubleshooting and Optimization Tips

• Solubility Issues: If precipitation occurs in water, apply ultrasonic assistance or switch to DMSO/ethanol for higher concentrations. Confirm solubility visually before use.
• Loss of Activity: Avoid repeated freeze-thaw cycles. Prepare small aliquots and use within one week when stored at -20°C. Discard any solutions showing discoloration or precipitation.
• Off-Target Effects: Validate specificity by including Y1R/NPFF knockout or knockdown controls. Consider dose-response experiments to establish a clear efficacy window.
• Batch Variation: Utilize the provided Certificate of Analysis (COA) and QC data (HPLC, MS, NMR) to verify batch consistency.
• Experimental Controls: In coculture systems, ensure cell-type purity and viability. Adjust compound dosing based on cell density and media composition to optimize receptor engagement.
• Signal Quantification: When assessing cAMP or downstream signaling, normalize data to total protein or cell count to control for experimental variability.

Future Outlook: Expanding the Reach of NPY/NPFF Pathway Interrogation

With the increasing recognition of the NPY/NPFF axis in diverse physiological and pathophysiological contexts, research enabled by BIBP 3226 trifluoroacetate is poised for rapid expansion. Potential directions include:

• High-Throughput Screening: Use in automated platforms to identify synergistic compounds modulating the neuropeptide Y receptor pathway for cardiovascular and neuropsychiatric indications.
• In Vivo Imaging: Coupling with radiolabeled ligands to visualize receptor occupancy and distribution in living systems.
• Personalized Medicine: Integrating NPY/NPFF system research with patient-derived stem cell models to tailor interventions for anxiety, pain, and arrhythmia.
• Cross-Talk Mechanisms: Examining interactions with other peptide systems (e.g., leptin, ghrelin) to elucidate integrated metabolic and neural regulation networks.

As advanced models such as coculture systems become standard in translational research, the demand for reliable, specific antagonists like BIBP 3226 trifluoroacetate will only grow. By bridging molecular pathways and cellular function, this compound remains a cornerstone for next-generation discovery in neuropeptidergic signaling.

For detailed product specifications, quality control documentation, and ordering information, visit the BIBP 3226 trifluoroacetate product page.