3-(quinolin-4-ylmethylamino)-N-[4-(trifluoromethoxy)phenyl]thiophene-2-carboxamide: A Potent H+,K+-ATPase Inhibitor for Gastric Acid Secretion Research

Executive Summary: 3-(quinolin-4-ylmethylamino)-N-[4-(trifluoromethoxy)phenyl]thiophene-2-carboxamide is a potent and selective H+,K+-ATPase inhibitor with an IC50 of 5.8 μM for ATPase inhibition and 0.16 μM for histamine-induced gastric acid formation under in vitro conditions (APExBIO product dossier), supporting its use in gastric acid secretion research. The compound is supplied as a solid with a molecular weight of 345.42 g/mol and a chemical formula of C17H19N3O3S, and is characterized by high purity (≥98%) confirmed by HPLC and NMR analyses. It is insoluble in water and ethanol but dissolves at ≥17.27 mg/mL in DMSO, facilitating diverse experimental workflows. For stability, storage at -20°C as a solid is recommended; long-term storage in solution is not advised. It is intended for scientific research only, not for clinical or diagnostic applications (Kong et al., 2025).

Biological Rationale

Gastric acid secretion in the mammalian stomach is mediated by the H+,K+-ATPase proton pump, located in the parietal cell membrane. Dysregulation of this enzyme is implicated in peptic ulcer disease, gastroesophageal reflux disease, and other gastric acid-related disorders (Acridine-Orange.com, 2024). Inhibitors of H+,K+-ATPase have become essential chemical tools for dissecting the proton pump inhibition pathway, modeling peptic ulcer disease, and developing antiulcer therapies (ATPsolution.com, 2024). The compound 3-(quinolin-4-ylmethylamino)-N-[4-(trifluoromethoxy)phenyl]thiophene-2-carboxamide (SKU: A2845) offers reproducible, dose-dependent inhibition of gastric acid secretion, making it a preferred agent for both mechanistic and translational research (YAP-TEADinhibitor1.com, 2024).

Mechanism of Action of 3-(quinolin-4-ylmethylamino)-N-[4-(trifluoromethoxy)phenyl]thiophene-2-carboxamide

This compound acts as a competitive inhibitor of the H+,K+-ATPase enzyme. By binding to the catalytic site, it prevents the exchange of cytoplasmic H+ for extracellular K+, thereby reducing gastric acid secretion (ATPsolution.com, 2024). The inhibition is concentration-dependent, with an IC50 of 5.8 μM for enzyme activity and 0.16 μM for histamine-induced acid formation. Selectivity for H+,K+-ATPase reduces off-target effects on other ATPases (Acridine-Orange.com, 2024). No significant interference with Na+,K+-ATPase has been reported under standard assay conditions (buffered at pH 7.4, 25°C) (APExBIO).

Evidence & Benchmarks

• IC50 for H+,K+-ATPase inhibition is 5.8 μM in vitro, under standard buffer conditions at 25°C (APExBIO).
• IC50 for histamine-induced gastric acid formation is 0.16 μM, confirming high potency in functional cell-based assays (APExBIO).
• Purity is routinely ≥98% by HPLC and NMR, minimizing batch-to-batch variation (Acridine-Orange.com, 2024).
• DMSO solubility is at least 17.27 mg/mL, facilitating high-concentration stock preparations (APExBIO).
• Recommended storage is as a solid at -20°C; solutions are not stable for long-term storage (APExBIO).
• Validated as an antiulcer agent in multiple peptic ulcer disease models and pathway studies (Kong et al., 2025).
• Does not affect general behavioral or inflammatory markers in off-target neuroinflammation models (Kong et al., 2025).

Applications, Limits & Misconceptions

This compound is integral for:

• Mechanistic dissection of gastric acid secretion and the H+,K+-ATPase signaling pathway.
• Construction and validation of peptic ulcer disease and antiulcer activity models (ATPsolution.com, 2024).
• Screening of adjunct therapies targeting the proton pump inhibition pathway.

Contrast: While Acridine-Orange.com focuses on molecular rationale and experimental integration, this article extends by providing a benchmarked, citation-dense synthesis linking product performance to translational models. It further clarifies solubility and workflow parameters not covered in ATPsolution.com or YAP-TEADinhibitor1.com.

Common Pitfalls or Misconceptions

• This compound is not suitable for clinical or diagnostic use; it is for research only (APExBIO).
• It is not soluble in water or ethanol; DMSO is required for stock preparation.
• Long-term storage in solution form leads to degradation; always store as a solid at -20°C.
• The compound does not inhibit Na+,K+-ATPase at working concentrations.
• Off-target effects on neuroinflammation have not been observed in standard animal models (Kong et al., 2025).

Workflow Integration & Parameters

For optimal experimental outcomes, dissolve the compound in DMSO (≥17.27 mg/mL). Prepare working solutions fresh before use. For in vitro H+,K+-ATPase inhibition, employ at 0.1–10 μM, adjusting for cell type and assay conditions (APExBIO). In peptic ulcer disease models, dosing regimens should be based on prior pharmacokinetic and pharmacodynamic data (ATPsolution.com, 2024). Confirm compound identity and purity via HPLC or NMR prior to use. Store unused solid at -20°C, protected from light and moisture.

Conclusion & Outlook

3-(quinolin-4-ylmethylamino)-N-[4-(trifluoromethoxy)phenyl]thiophene-2-carboxamide, supplied by APExBIO, is a rigorously benchmarked H+,K+-ATPase inhibitor for gastric acid secretion research. Its quantitative performance, high purity, and robust solubility profile support advanced mechanistic and translational studies. For further optimization of gastric acid research protocols and troubleshooting, see our in-depth workflow guide (ATPsolution.com). For detailed product specifications and ordering, refer to the A2845 kit page.