NPP1-IN-2 , CAS No.N1438753

CAS: N1438753 Cat. No.: N1438753 Fórmula: C₂₂H₂₂N₄OS Peso molecular: 390.50

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Descripción general

NPP1-IN-2 (compound 3h) is a potent and selective Ectonucleotide pyrophosphatase/phosphodiesterase-1 (NPP1) inhibitor with IC 50 values of 0.55, 3.73 µM for NPP1, NPP3, respectively.

Specifications

Condiciones de almacenamiento de almacenamiento

Store at -20°C

Enviado en

Ice chest + Ice pads

Este producto requiere envío en cadena de frío. Los servicios terrestres y otros servicios económicos no están disponibles.

Tipo de acción

INHIBITOR

Nombres e identificadores

Peso molecular

390.50

Application Protocols

Item-specific, validated protocols are not provided. The following general workflows are commonly used for small-molecule ENPP1 inhibitors (literature/practice; not item-specific):

• DMSO stock preparation:
  • Warm to room temperature in a desiccator before opening. Prepare 10–50 mM stocks in anhydrous DMSO. Vortex to dissolve, then clarify by brief centrifugation. Aliquot into low-bind tubes and store at -20°C.
• Biochemical inhibition assay (fluorescent or HPLC endpoint):
  • Buffer: Tris or HEPES, pH 7.4–8.0, with NaCl (100–150 mM) and required divalent cations (e.g., Zn2+, Mg2+) per enzyme source.
  • Add enzyme to buffer, preincubate with inhibitor (constant DMSO, typically ≤1%). Initiate with substrate (e.g., nucleotide phosphodiester). Incubate at 25–37°C. Quench or read kinetically.
  • Determine IC50 via 10-point serial dilution (1:3 or 1:2 steps). Fit using a four-parameter logistic model; report mean ± SD over ≥3 replicates.
• Cell-based assays:
  • Verify cell tolerance to DMSO and compound. Assess pathway modulation with appropriate biomarkers. Include genetic controls and counterscreens.
• Orthogonal confirmation:
  • Confirm inhibition by an alternative detection method (e.g., switch from fluorescence to LC–MS) to rule out reporter interference.

Adjust parameters to your specific enzyme preparation and detection modality.

Biological Roles

This section summarizes target biology relevant to an ENPP1 (NPP1) inhibitor; it does not ascribe biological properties to this specific item beyond its intended use as a research probe.

• ENPP1 function (literature): Ectonucleotide pyrophosphatase/phosphodiesterase 1 is a membrane-associated ectoenzyme that hydrolyzes extracellular nucleotide phosphodiesters and pyrophosphates. Canonical activities include ATP→AMP + PPi and hydrolysis of 2′,3′-cGAMP to AMP + GMP. ENPP1 thereby regulates extracellular nucleotide pools, mineralization via PPi, and modulates the cGAS–STING signaling axis by degrading cGAMP.
• Localization: Predominantly extracellular/pericellular; contains a catalytic site coordinated by divalent metals (commonly Zn2+ in literature models).
• Research utility of inhibitors (general): Chemical inhibition allows temporal control to dissect ENPP1’s role in nucleotide turnover, signaling crosstalk, and interactions with other ectoenzymes (e.g., CD39, CD73). Inhibitors are used in biochemical assays, live-cell target engagement studies, and pathway mapping with genetic complementation as controls.
• Selectivity considerations: ENPP family members (ENPP1–7) have overlapping but distinct substrate preferences; off-target inhibition can confound interpretation. Counterscreens and rescue experiments are recommended.

Note: No pharmacological claims are made for this product. It is provided strictly for research use to study enzyme biology and related pathways.

Buffer Applications

This product is a small-molecule research inhibitor and is not itself a buffering agent. However, buffer composition is critical for ENPP1 enzymology and inhibitor characterization (literature guidance; not item-specific):

• Buffer systems: Tris or HEPES, pH 7.2–8.0, are commonly used for ENPP1 assays. Ionic strength is typically maintained with NaCl (e.g., 100–150 mM) to approximate physiological conditions.
• Metal ions: ENPP1 activity depends on divalent cations in the active site; assay buffers often include Zn2+ and/or Mg2+ (concentrations vary in literature). Metal chelators (e.g., EDTA) can suppress activity and confound results.
• Detergents and carriers: 0.01–0.05% nonionic surfactant (Tween-20 or Pluronic F-127) can reduce nonspecific adsorption and compound aggregation. 0.1% BSA is sometimes included as a carrier to reduce sticking.
• DMSO tolerance: Validate enzyme activity across the DMSO range you plan to use (e.g., 0–1% v/v), because some enzymes are solvent-sensitive.
• Controls: Include buffer-only, substrate-only, and vehicle controls to establish baseline activity and signal stability over the assay time course.

Always optimize buffer for the specific substrate and detection method (fluorescent reporters, HPLC/LC–MS, or inorganic phosphate/pyrophosphate readouts).

Green Alternatives

With unknown structure and properties, green chemistry considerations focus on formulation and assay design rather than the molecule itself.

Greener handling choices (general guidance):

• Solvent minimization: Use the lowest feasible DMSO percentage in assays (e.g., ≤0.5% v/v) to reduce solvent burden while maintaining solubility.
• Solvent selection: Prefer DMSO over DMF where possible—DMSO is lower volatility, lower flammability, and generally considered a more benign polar aprotic option in small volumes for screening.
• Waste reduction: Prepare concentrated master stocks and dispense by acoustic or low-volume liquid handling to minimize disposable plastic and solvent waste.
• Energy use: Store at -20°C as specified, but avoid unnecessary deep-freeze cycling by aliquoting single-use volumes.
• Data-driven design: Early solubility screening in aqueous buffers can reveal opportunities to run assays with reduced organic cosolvent.

Illustrative comparison (general; not item-specific):

• DMSO (preferred for stocks): high solvency, low VOC emissions, miscible with water; disposal as aqueous-organic waste.
• Ethanol: renewable source potential; higher volatility increases emissions; may perturb enzyme activity at modest percentages.
• DMF/Acetonitrile: strong solvents but higher hazard profiles; avoid for routine stocks unless specifically required by the chemotype.

Note: For process-scale green assessments (unlikely for a screening probe), full structural knowledge is required to evaluate lifecycle and hazard metrics.

Pharmaceutical Uses

No pharmacopeial status, excipient role, or formulation specifications are provided for this product. It is supplied strictly for research use only.

Context for discovery workflows (general guidance; not item-specific):

• Hit-to-lead support: Small-molecule ENPP1 inhibitors are employed in early discovery to generate SAR, enzyme kinetics, and cell-based pathway data. Compounds may be profiled for microsomal stability, permeability, and solubility as part of preclinical research; such data, if available, would be lot- and scaffold-specific and are not provided here.
• Formulation for research dosing (preclinical labs): When in vivo or ex vivo research is contemplated, investigators often explore solubilization in biocompatible vehicles (e.g., PEG/solutol/saline systems). Any such use requires institution-specific review and is outside the scope of this listing.
• Regulatory: Not intended for human or veterinary use. No GMP status is implied. If regulated studies require defined quality attributes (e.g., residual solvents, elemental impurities), contact us regarding available analytical support.

For any application beyond routine in vitro research, obtain structural, analytical, and stability information for the exact lot via CoA/SDS and perform independent qualification.

Physical Properties

Item-specific physico-chemical properties are not provided in the Product Data.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Purity/grade: Not specified for this item; refer to CoA/Spec Sheet.
• Melting point, boiling point, density, refractive index, UV cut-off: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility (item-specific): Not specified for this item; refer to CoA/Spec Sheet.

General guidance for small-molecule screening compounds (literature/practice; not item-specific):

• Many ENPP1 inhibitor chemotypes are moderately lipophilic and are routinely prepared as concentrated DMSO stocks (e.g., 10–50 mM) and then diluted into aqueous assay buffers containing 0.01–0.1% nonionic surfactant to mitigate aggregation and adsorption.
• Typical screening buffers (literature) for ecto-nucleotide phosphodiesterases are near-neutral pH (7.2–8.0) and may include divalent cations (e.g., Zn2+, Mg2+), which can influence apparent potency.
• Ensure test concentrations remain below solubility limits; verify by visual inspection for precipitation and by parallel LC–UV/LC–MS after dilution.
• For spectroscopy-based assays, check compound absorbance/fluorescence in the detection window to avoid inner-filter effects; collect a reference UV–Vis profile when available.

Note: For all quantitative property needs (logP, pKa, aqueous solubility), please consult literature for the specific scaffold or request analytical data tied to the supplied lot.

Quality and Grades

• Grade/purity: Not specified for this item; refer to CoA/Spec Sheet.

Context and expectations for small-molecule screening compounds (general guidance):

• Research-use screening grade typically prioritizes identity confirmation (e.g., LC–MS) and a chromatographic purity suitable for biological assays (often >95% by HPLC/UPLC). Actual specification must be confirmed on the item’s CoA.
• Analytical package: For hit validation and SAR, researchers often request lot-specific LC–MS traces, purity by HPLC (single wavelength and/or PDA), and, where necessary, 1H NMR. Please contact us to obtain the current lot’s analytical set.
• Stabilizers: Not specified for this item. If present in a given lot, stabilizers or counter-ions will be disclosed on the CoA/SDS.
• Low-UV/low-fluorescence requirements (assay compatibility): If your detection modality is UV/fluorescence-based, request PDA spectra to ensure minimal interference at assay wavelengths.
• Trace contaminants and residual solvents: Not specified for this item; refer to CoA/Spec Sheet. If critical, we can provide residual solvent screening per ICH Q3C on request.
• Lot-to-lot control: Retain and reference the SKU (N1438753) and lot number; when repeating biological data, match lot numbers or re-qualify activity after any lot change.

Reaction and Applications

As a designated screening inhibitor (NPP1-IN-2), this product is intended for biochemical and cell-based research rather than for use as a synthetic reagent. No item-specific application notes are provided.

Research applications (general, target-focused; literature guidance):

• ENPP1 enzymology: Probe ENPP1-mediated hydrolysis of nucleotide phosphodiester/pyrophosphate substrates (e.g., ATP→AMP+PPi; cGAMP→AMP+GMP). Assess inhibition using colorimetric, fluorescent, HPLC, or LC–MS readouts.
• Pathway interrogation: Modulate extracellular nucleotide metabolism and cGAS–STING pathway signaling in cell models by inhibiting ENPP1 activity; confirm on-target effects with genetic controls.
• Selectivity profiling: Run counterscreens against related ENPP family members (e.g., ENPP2/autotaxin) and unrelated phosphodiesterases to characterize off-target activity.
• Mechanistic studies: Determine competitive vs noncompetitive behavior via Michaelis–Menten kinetics and substrate titrations across fixed inhibitor levels.

Practical tips (general):

• Verify solubility at working concentration and assay temperature. Centrifuge or filter pre-dilutions to remove particulates.
• Include DMSO-matched vehicle controls; maintain identical final solvent percentages across all wells.
• Check time-dependence of inhibition (preincubation experiments) to detect slow-binding or covalent mechanisms.
• Confirm activity with orthogonal assay formats to de-risk reporter interference.

Reaction Conditions

Not applicable in the classical synthetic sense: this product is a screening inhibitor, not a reagent or catalyst. No reaction condition specifications are provided.

Assay-condition guidance (general; not item-specific):

• Temperature: Many ENPP1 biochemical assays are conducted at 25–37°C. Verify thermal stability of both enzyme and compound at the chosen temperature.
• Preincubation: A 5–30 min preincubation of inhibitor with enzyme can reveal slow-onset inhibition; maintain constant DMSO content across conditions.
• Substrate concentration: Use near the Km for mechanistic analyses; run substrate titrations to classify competitive/noncompetitive behavior.
• Time course: Ensure linear product formation over the measurement window. Quench reactions appropriately (e.g., with chelators/acid for HPLC endpoints) before analysis.
• Replicates and controls: Include technical replicates and orthogonal readouts (e.g., fluorescent reporter vs LC–MS) to exclude assay interference.

All numeric parameters above are literature-style guidance and must be optimized for your enzyme preparation, substrate, and detection method.

Safety and Handling

• GHS classification, signal word, hazard and precautionary statements: Not specified for this item; refer to the SDS.
• General laboratory PPE: lab coat, safety glasses, and appropriate chemical-resistant gloves (e.g., nitrile). Work in a chemical fume hood to avoid inhalation of dusts/aerosols.
• Storage conditions (item-specific): Store at -20°C (per Product Data). Protect from moisture and light. Keep container tightly closed.
• Shipping: Ice chest + ice pads (per Product Data). Allow to equilibrate to room temperature in a desiccator before opening to minimize moisture condensation.
• Incompatibilities (general): Avoid strong oxidizers and strong acids/bases unless compatibility is known for the specific chemotype. Do not heat to decomposition; decomposition products are unknown.
• First aid (general overview; defer to SDS):
  • Inhalation: Move to fresh air; seek medical attention if symptoms persist.
  • Skin contact: Wash with soap and water; remove contaminated clothing.
  • Eye contact: Rinse cautiously with water for several minutes; remove contact lenses if present and easy to do; continue rinsing.
  • Ingestion: Rinse mouth; seek medical advice.
• Hygiene measures: Avoid contact with skin and eyes. Do not pipette by mouth. Wash hands after handling. Prevent environmental release.
• Note: This product is for research use only. Consult the official SDS for authoritative hazard classification and response.

Solvent Selection

This product is supplied as a small-molecule research inhibitor; item-specific solvent data are not provided.

General solvent guidance for screening compounds (literature/practice; not item-specific):

• Primary stock solvent: DMSO is the standard for preparing concentrated stocks (e.g., 10–50 mM) owing to broad solvency and good miscibility with aqueous buffers at low final percentages.
• Aqueous assay media: Dilute DMSO stocks into Tris, HEPES, or PBS-based buffers. Maintain final DMSO typically ≤0.5–1% v/v to limit effects on enzyme activity and cell viability.
• Surfactants: 0.01–0.05% Tween-20 or Pluronic F-127 can reduce nonspecific adsorption and aggregation artifacts in biochemical screens (literature guidance).
• Alternative cosolvents: If DMSO is unsuitable, consider DMF or ethanol (cautiously, due to volatility and enzyme sensitivity). Validate activity and solubility after any solvent change.
• Light and oxygen sensitivity: If the chemotype is unknown, handle under subdued light and minimize air exposure until stability is established by analytical checks.

Comparison (general):

• DMSO: highest solvency, low volatility; common choice for libraries.
• Ethanol: volatile, lower solvency; useful for rapid evaporation formats.
• Aqueous-only: preferred for immediate-use solutions when solubility allows; verify clarity and stability over the assay time course.

Storage and Reconstitution

• Storage (item-specific): Store at -20°C (per Product Data). Keep container tightly closed, protected from light and moisture. Minimize headspace and freeze–thaw cycles.
• Shipping: Ice chest + ice pads (per Product Data). Inspect upon receipt; if thawed, re-freeze promptly after aliquoting.
• Reconstitution (general guidance; not item-specific):
  • For screening use, prepare concentrated stocks in anhydrous DMSO (e.g., 10–50 mM). Mix until fully dissolved; if necessary, sonicate briefly. Filter if particulate persists and compatibility is confirmed.
  • Aliquot into single-use portions (low-bind polypropylene) to avoid repeated freeze–thaw. Store aliquots at -20°C. Record preparation date and concentration.
  • For aqueous working solutions, dilute the DMSO stock into pre-warmed assay buffer under vigorous mixing to avoid local supersaturation. Keep final DMSO ≤0.5–1% v/v unless otherwise validated.
• Stability: Item-specific stability data are not provided; refer to CoA/Spec Sheet. Verify stability by periodic LC–MS/HPLC of stored stocks.
• Disposal: Dispose of solutions and materials according to your institution’s chemical waste procedures.

Note: For any deviations (alternative solvents, higher temperatures), confirm compatibility experimentally and document outcomes for reproducibility.

Structure and Identity

• Item name: NPP1-IN-2 (catalog probe; small-molecule inhibitor intended for research screening against ENPP1)
• CAS: N1438753 (catalog placeholder; not a registered CAS number in public databases)
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey: Not specified for this item; refer to CoA/Spec Sheet.
• Structural features: Not provided in the Product Data. Based on the name, this product is categorized as an ENPP1 (NPP1) inhibitor; specific functional groups, ring systems, and stereochemistry are not disclosed in this listing.
• 2D structure description: Not available in this entry.
• Identity notes:
  • If you require definitive structure-related identifiers (SMILES/InChIKey), please request the current CoA/SDS or specification sheet.
  • For screening-library management, track identity via SKU (N1438753) and lot number to align analytical data (LC–MS, HPLC, NMR) with your internal records.

Synthetic Utility

This item is offered as a finished small-molecule inhibitor for biochemical research and is not intended as a synthetic building block. No reactive handles or functional-group details are provided.

General notes (when considering derivatization; not item-specific):

• Without a disclosed structure, planning covalent attachment (e.g., to beads or probes) or analogue synthesis is not feasible. If you require a derivatizable analogue (amine/azide/alkyne handle), please inquire about custom synthesis.
• For chemical biology applications (e.g., affinity probes), structure is essential to position linkers away from binding pharmacophores; SAR and co-crystal information (if available in literature for related scaffolds) guide such designs.
• Stability and reactivity: Unknown for this specific item; avoid exposure to strong acids/bases and oxidants until stability is established analytically.

Recommendation: Use this product as supplied for target engagement studies. For synthetic routes or intermediates, request a proposal for a custom route once structural disclosure is available under CDA/NDA as appropriate.

Target Specificity

• Target, clone/epitope, isotype, and species reactivity: Not applicable; this is a small-molecule inhibitor, not an antibody or protein reagent.

General considerations for small-molecule selectivity (not item-specific):

• Family profiling: Evaluate activity against ENPP family members (ENPP1–7), with special attention to ENPP2 (autotaxin), to quantify selectivity.
• Off-target panels: Consider broader phosphodiesterase and hydrolase panels, and a safety pharmacology set (e.g., hERG, CYPs) if relevant to your research.
• Mechanistic probes: Use substrate-competition experiments and mutationally altered ENPP1 constructs to confirm binding mode.

Item-specific potency/selectivity data are not provided in this listing; consult the CoA/Spec Sheet or generate de novo profiles under your assay conditions.