BIBP 3226 trifluoroacetate: Precision NPY Y1/NPFF Antagonist for NPY/NPFF System Research

Executive Summary: BIBP 3226 trifluoroacetate (CAS: 1068148-47-9) is a highly selective, non-peptide antagonist of neuropeptide Y Y1 (NPY Y1) and neuropeptide FF (NPFF) receptors, with Ki values of 1.1 nM (rat NPY Y1), 79 nM (human NPFF2), and 108 nM (rat NPFF) under standardized binding assays (APExBIO). It competitively inhibits NPFF-induced suppression of forskolin-stimulated cAMP in vitro and blocks NPFF-mediated hypothermic and anti-opioid effects in rodents (Fan et al., 2024). The compound enables targeted investigation of the NPY/NPFF axis in advanced coculture systems modeling cardiovascular and neurobiological disorders. Recent research confirms the pivotal role of the NPY/Y1R pathway in cardiac arrhythmogenesis, highlighting the need for selective pharmacological tools (Fan et al., 2024). BIBP 3226 trifluoroacetate is supplied by APExBIO with validated purity (>98%), ensuring reproducibility and integrity in NPY/NPFF system research.

Biological Rationale

The neuropeptide Y (NPY) and neuropeptide FF (NPFF) systems regulate key physiological processes, including anxiety modulation, pain processing, and cardiovascular function (Fan et al., 2024). The NPY Y1 receptor (Y1R) is a G protein-coupled receptor (GPCR) predominantly expressed in the brain, heart, and peripheral tissues. Its activation by NPY influences cell signaling through cyclic AMP (cAMP) pathways. NPFF receptors, including NPFF1 and NPFF2, are implicated in opioid modulation and thermoregulation. Recent evidence demonstrates that the adipose-neural axis, mediated by leptin and NPY/Y1R signaling, contributes to cardiac arrhythmias via enhanced sympathetic neuron activation and altered myocardial electrophysiology (Fan et al., 2024). Model systems using cocultures of adipocytes, sympathetic neurons, and cardiomyocytes facilitate mechanistic dissection of these pathways (Dissecting the Adipose-Neural Axis). BIBP 3226 trifluoroacetate, as a potent and selective antagonist, is essential for experimentally defining the roles of these neuropeptide pathways in health and disease.

Mechanism of Action of BIBP 3226 trifluoroacetate

BIBP 3226 trifluoroacetate acts as a competitive antagonist at the NPY Y1 and NPFF receptors. It binds with high affinity to the rat NPY Y1 receptor (Ki = 1.1 nM), human NPFF2 receptor (Ki = 79 nM), and rat NPFF receptor (Ki = 108 nM), established via radioligand binding assays at 25°C in buffered solutions (APExBIO). By occupying the orthosteric site on Y1 and NPFF receptors, BIBP 3226 prevents endogenous ligands from activating downstream G protein signaling. In cellular assays, it blocks NPFF-induced inhibition of forskolin-stimulated cAMP production, indicating antagonism of Gi/o-coupled receptor pathways. In vivo, BIBP 3226 trifluoroacetate abolishes NPFF-dependent hypothermic and anti-opioid responses in rodent models, confirming functional antagonism (Fan et al., 2024). The compound does not exhibit significant off-target activity at other GPCRs at concentrations up to 1 μM, as demonstrated in selectivity panels (APExBIO).

Evidence & Benchmarks

• BIBP 3226 trifluoroacetate exhibits high binding affinity for rat NPY Y1 (Ki = 1.1 nM), human NPFF2 (Ki = 79 nM), and rat NPFF (Ki = 108 nM) in competitive binding studies (APExBIO).
• In coculture models, NPY/Y1R blockade by BIBP 3226 prevents arrhythmogenic signaling induced by adipocyte-derived leptin and NPY in cardiomyocytes (Fan et al., 2024).
• BIBP 3226 reverses NPFF-induced inhibition of forskolin-stimulated cAMP production, confirming antagonism of Gi/o-linked signaling (BIBP 3226: Tool for NPY/NPFF System Dissection).
• Administration in rodents blocks NPFF-mediated hypothermic and anti-opioid effects, validating in vivo efficacy (Fan et al., 2024).
• Compound has >98% purity, with batch validation by HPLC, MS, and NMR for every lot (APExBIO).

This article extends the mechanistic coverage provided in BIBP 3226: Precision Tool for NPY/NPFF System Dissection by integrating new data from advanced coculture models and cardiac arrhythmia research.

Applications, Limits & Misconceptions

BIBP 3226 trifluoroacetate is validated for use in physiological, pharmacological, and mechanistic studies targeting the NPY/NPFF system. It is particularly valuable in:

• Cardiac arrhythmia models exploring adipose-neural signaling, as established in stem cell-based coculture systems (Fan et al., 2024).
• Analgesia research, where NPFF antagonism elucidates opioid modulation (BIBP 3226: Experimental Rigor in NPY/NPFF Studies).
• Anxiety models, dissecting the contributions of NPY/Y1R pathways to stress responses.
• In vitro and ex vivo systems requiring selective, non-peptide antagonism without peptide degradation concerns.

Common Pitfalls or Misconceptions

• BIBP 3226 trifluoroacetate is not a general NPY receptor blocker; it is highly selective for Y1 and NPFF subtypes, with minimal activity at Y2, Y4, or Y5 (APExBIO).
• The compound is for in vitro and in vivo research only; it is not approved for diagnostic or therapeutic use.
• Long-term storage of prepared solutions is not recommended due to possible activity loss; solutions should be freshly prepared (APExBIO).
• Solubility may require ultrasonic assistance in aqueous buffers; exceeding recommended concentrations may lead to precipitation or reduced efficacy.
• Off-target effects may occur at concentrations above 1 μM; always use validated working concentrations.

Workflow Integration & Parameters

BIBP 3226 trifluoroacetate is supplied as an off-white solid, with a molecular weight of 587.59 Da and chemical formula C29H32F3N5O5. It is soluble at ≥78 mg/mL in DMSO, ≥73.2 mg/mL in ethanol, and ≥12.13 mg/mL in water (with ultrasonic assistance) at 25°C. For most cell-based assays, working concentrations range from 1 nM to 1 μM, with vehicle controls recommended. Solutions should be freshly prepared and used promptly to prevent loss of activity. Storage at -20°C is advised for the solid form; avoid repeated freeze-thaw cycles. Each purchase from APExBIO (B7155 kit) is accompanied by a certificate of analysis, HPLC chromatogram, and NMR/MS verification (APExBIO). For trouble-shooting strategies and advanced workflow integration, see the guidance in BIBP 3226: Unveiling NPY/NPFF System Dissection, which this article updates by including data from 2024 cardiac arrhythmia models.

Conclusion & Outlook

BIBP 3226 trifluoroacetate is a standard-setting tool for dissecting the NPY/NPFF system in anxiety, analgesia, and cardiovascular research. Its high selectivity and validated performance in advanced models support robust mechanistic studies. Recent findings in epicardial adipose tissue–related cardiac arrhythmias underscore the relevance of NPY/Y1R antagonists in translational research (Fan et al., 2024). Continued integration of BIBP 3226 into multi-system models will accelerate discovery in neuro-adipose signaling. For ordering and quality documentation, visit the product page at APExBIO.