TrkA-IN-4 - ≥99% , CAS No.T658654

CAS: T658654 Cat. No.: T658654 Formula: C₂₇H₂₁F₃N₄O₅ Peso molecolare: 538.47

QUALITÀ & PUREZZA ≥99%

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

TrkA-IN-4, a potent, orally active and allosteric TrkA inhibitor, is a proagent of TrkA-IN-3 ( HY-151948 ). TrkA-IN-4 exhibits potent antinociceptive effects.

Appearance:Solid

IC50& Target:TrkA

In Vitro:TrkA-IN-4 (compound 39) demonstrates 65.1% and 46.3% of kinase inhibition towards TrkA at concentrations of 1 μM and 0.1 μM, respectively. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

In Vivo:TrkA-IN-4 (compound 39) (0.9375-120mg/kg; i.g.) exhibits stronger maximum antinociceptive effects 3 h after administration, with an ED 50 of 7.836mg/kg in hot plate testing on male mice. MCE has not independently confirmed the accuracy of these met

Biological Activity:TrkA-IN-4, a potent, orally active and allosteric TrkA inhibitor, is a proagent of TrkA-IN-3 ( HY-151948 ). TrkA-IN-4 exhibits potent antinociceptive effects.

Specifiche

Specifiche e purezza

≥99%

Condizioni di conservazione di stoccaggio

Store at -20°C

Spedito in

Ice chest + Ice pads

Questo prodotto richiede spedizione a catena fredda. I servizi di terra e altri servizi economici non sono disponibili.

Purezza

≥99%

Nomi e identificatori

Sorrisi canonici

CC(OCOC(C1=CN=C(C(NC(C2=CC=C(C(C3=NN(C=C3)C)=C2)C(F)(F)F)=O)=C1)C4=CC=CC=C4)=O)=O

Peso molecolare

538.47

Proprietà chimiche e fisiche

• Solubilità: DMSO : 50 mg/mL (92.86 mM; ultrasonic and warming and heat to 60°C)

Application Protocols

• Item-specific tested applications/dilutions: Not specified for this item; refer to CoA/Spec Sheet.

• Example protocols (general guidance; adjust to your system)

  • Enzyme inhibition (ADP-Glo format)
    • Prepare 10 mM DMSO stock; serially dilute 1:3–1:5 in DMSO to generate a 10-point curve.
    • Pre-incubate TrkA enzyme with inhibitor for 15–30 min at room temperature in assay buffer (e.g., 50 mM HEPES pH 7.5, 10 mM MgCl2, 0.01% Tween-20, 1 mM DTT).
    • Initiate with ATP/substrate; incubate 30–60 min; develop and read luminescence. Fit IC50 with appropriate models accounting for ATP concentration.
  • Cellular phospho-TrkA readout (WB/ELISA)
    • Seed cells expressing TrkA; serum-starve as needed.
    • Treat with inhibitor (e.g., 0.01–10 μM final; constant DMSO) for 30–60 min; stimulate with NGF.
    • Lyse cells; quantify p-TrkA (Tyr490) and downstream markers (e.g., p-ERK) with normalization to total protein.
  • Thermal shift (DSF)
    • Mix purified TrkA with dye (e.g., SYPRO Orange) and inhibitor at varying concentrations; ramp temperature; analyze Tm shifts to infer binding.

• Controls

  • Include a reference Trk inhibitor as positive control and vehicle-only negative control. Replicate across plates to assess robustness (Z′-factor).

Biological Roles

• Target context (general literature; not item-specific claims)

  • TrkA (NTRK1) is a receptor tyrosine kinase activated by nerve growth factor (NGF), initiating downstream signaling through MAPK/ERK, PI3K/AKT, and PLCγ pathways.
  • Small-molecule TrkA inhibitors are used as chemical tools to dissect NGF/TrkA signaling, study receptor phosphorylation dynamics, and parse pathway crosstalk with TrkB/TrkC.

• Research utility of TrkA chemical probes (general)

  • Enable temporal control of pathway inhibition in cells and tissues, complementing genetic knockdown/knockout approaches.
  • Facilitate structure–activity relationship (SAR) studies when screened across Trk family variants and mutants.
  • Support target engagement measurements via thermal shift (DSF/CETSA) and phospho-proteomics.

• Note on biological activity

  • Potency, selectivity, and cellular permeability for TrkA-IN-4 are not specified for this item; consult CoA/Spec Sheet and primary references where available.
  • All uses are for research purposes only. No medical or clinical claims are made.

Buffer Applications

• This product is not a buffering agent. However, buffers are integral to its use in biochemical assays (general guidance):

  • Kinase assay buffers commonly contain HEPES or Tris (pH 7.0–7.5), Mg2+ (e.g., 5–10 mM MgCl2), ATP (varied to probe competition), and reducing agents (e.g., 1–2 mM DTT) as appropriate.
  • For cell-based studies, use physiologically compatible media and serum conditions; maintain constant DMSO across treatments.
  • Include detergent (e.g., 0.01–0.05% Tween-20) to reduce nonspecific adsorption in microplates when needed.

• Item-specific buffer compatibility: Not specified for this item; verify empirically and consult CoA/SDS.

Green Alternatives

• As a screening compound, the concept of a greener alternative pertains primarily to solvent and process choices during handling and assay preparation rather than substitution of the active entity.

• Greener solvent considerations (general guidance)

  • Prefer aqueous buffers with minimal DMSO (e.g., 0.1–0.5% v/v) where solubility permits.
  • When organic dissolution is required, consider ethanol or propylene carbonate as alternatives to DMF/NMP if compatible with your assay; verify solubility and biological tolerance.
  • For purification (if needed), replace acetonitrile-rich mobile phases with methanol where chromatographic performance allows; employ water-rich gradients.

• Waste minimization

  • Prepare the smallest feasible volumes of concentrated stock solutions to limit DMSO waste.
  • Use microscale assays (384/1536-well formats) to reduce solvent consumption.

• Comparison snapshot (general)

  • DMSO: Excellent solubilizer; moderate environmental impact; widely accepted in assays.
  • DMF/NMP: Powerful solvents; higher regulatory scrutiny and worker exposure concerns—avoid when possible.
  • Ethanol/MeOH: More benign but may limit solubility and can perturb biological systems at lower concentrations.

Pharmaceutical Uses

• This product is supplied strictly for research use only.
• No pharmacopeial status, excipient role, or manufacturing specifications are provided for this item.
• While small-molecule TrkA inhibitors are of interest as research tools in drug discovery, no formulation or clinical use information is claimed or provided here for TrkA-IN-4.
• Any development toward dosage forms would require separate studies (solubility, stability, polymorph screening, salt selection), which are outside the scope of this listing.

Physical Properties

• Item-specific values

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.
  • Melting point, boiling point, density, refractive index, UV cutoff, water/peroxide/metal content: Not specified for this item; refer to CoA/Spec Sheet.

• General expectations for small-molecule kinase inhibitors (literature/general, not item-specific)

  • Physical state: Frequently supplied as off-white to yellow crystalline solids or amorphous powders.
  • Solubility: Often sparingly soluble in water; typically soluble in polar aprotic organics such as DMSO and DMF, and variably in methanol/ethanol and acetonitrile.
  • Ionization: Many TrkA-site binders are weakly basic or neutral; salt forms may be prepared to improve aqueous handling.

• Practical notes

  • For analytical characterization, LC–MS with water/acetonitrile + 0.1% formic acid and UV detection at 210–280 nm is commonly effective for heteroaromatic inhibitors (general guidance).
  • If solid, differential scanning calorimetry (DSC) can aid in assessing thermal behavior and polymorphism (general guidance).

Quality and Grades

• Item-specific grade/purity

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

• Guidance on grades for screening compounds (general information)

  • Research-grade small molecules are typically supplied at ≥95% purity by HPLC for biochemical/cellular screening. Higher purities (≥98%) can reduce assay background and off-target artifacts.
  • Documentation: A Certificate of Analysis (CoA) commonly includes identity confirmation (1H NMR and/or HRMS), purity by HPLC/UPLC, and sometimes water content by Karl Fischer.

• Stabilizers and form

  • Not specified for this item; refer to CoA/Spec Sheet. Inhibitors are generally supplied neat as solid or as a defined salt. If a stabilizer is present, it will be listed on the CoA/SDS.

• What to verify upon receipt (best practices)

  • Check lot-specific purity, counterion (if salt form), and assay for polymorph/solvate if relevant to your application.
  • Verify solubility in your assay vehicle (e.g., DMSO) and absence of particulates by visual inspection and test injections on LC before screening.

Reaction and Applications

• This product is a small-molecule screening compound and is not typically used as a synthetic reagent or catalyst.

• Research applications (general guidance; not item-specific claims)

  • Target class: Tropomyosin receptor kinase A (TrkA, NTRK1) inhibitor candidate for biochemical enzymology, kinase profiling, and mechanistic studies.
  • Representative assay types: ATP-competitive kinase activity assays (radiometric, ADP-Glo, mobility shift), binding assays (thermal shift/DSF, CETSA), and cell-based pathway readouts (e.g., NGF-stimulated TrkA phosphorylation level by Western blot or ELISA).
  • Selectivity assessment: Evaluate against TrkB/TrkC and a kinome panel to quantify off-target activity (general practice for kinase probes).

• Practical notes

  • Maintain final DMSO concentration consistently across assay wells (commonly ≤0.5–1.0% v/v) to avoid solvent artifacts.
  • Use serial dilutions spanning at least 3–4 orders of magnitude to define IC50 with confidence; include appropriate positive/negative controls and a reference inhibitor.
  • Confirm reversibility/ATP-competition via shift in IC50 with ATP concentration (general mechanistic check).

Reaction Conditions

• Not typically applicable: TrkA-IN-4 is a screening compound, not a reagent for chemical transformations.

• General handling/solution preparation conditions for assays (guidance)

  • Stock solutions: Prepare at 10–50 mM in anhydrous DMSO. Vortex and, if needed, gently warm (≤40 °C) to aid dissolution; avoid prolonged heating.
  • Working concentrations: Commonly 0.1 nM–50 μM in biochemical assays; 10 nM–10 μM in cell assays, depending on potency and cytotoxicity of the specific compound (ranges are general literature values, not item-specific).
  • Vehicle limits: Keep final DMSO ≤0.5–1.0% v/v in biological systems to minimize solvent effects.
  • Filtration: Optional 0.22 μm PTFE filtration for clarity if compatible; avoid binding-prone membranes for hydrophobic compounds.
  • Light/moisture: Prepare fresh aqueous dilutions immediately before use; protect from strong light if chromophoric groups are suspected.

• For any genuine chemical reactions using this molecule (e.g., conjugation), consult primary literature after confirming structure; no item-specific reaction data are provided.

Safety and Handling

• Item-specific hazard data

  • Signal Word: Not specified for this item; refer to SDS.
  • H-Statements: Not specified for this item; refer to SDS.
  • GHS Classification/Pictograms: Not specified for this item; refer to SDS.

• General laboratory precautions (general guidance; always defer to SDS)

  • Handle in a chemical fume hood; avoid inhalation of dust/aerosols and contact with skin or eyes.
  • Recommended PPE: lab coat, safety glasses, and appropriate chemical-resistant gloves (e.g., nitrile). Use respiratory protection if dusts/vapors cannot be controlled.
  • Avoid ingestion and environmental release. Collect spills with inert absorbent; dispose according to institutional and local regulations.

• Incompatibilities and stability (general)

  • Unknown for this specific item; check SDS. As with many heteroaromatic small molecules, avoid strong oxidizers and strong acids/bases during storage and handling unless compatibility is confirmed.
  • Protect from light and moisture as a precaution. Minimize repeated freeze–thaw of DMSO stocks.

• First-aid overview (general)

  • Skin/eye contact: Rinse with water for at least 15 minutes; remove contaminated clothing; seek medical attention as needed.
  • Inhalation: Move to fresh air; seek medical attention if symptoms persist.
  • Ingestion: Rinse mouth; do not induce vomiting; seek medical attention.

• Authoritative source: Always consult the product’s SDS for definitive safety information.

Solvent Selection

• Item-specific solubility: Not specified for this item; refer to CoA/Spec Sheet.

• General solvent guidance for hydrophobic kinase inhibitors (literature/general)

  • Primary stock solvent: DMSO is the standard due to excellent solvating power for aromatic heterocycles and compatibility with most biochemical and cell assays.
  • Co-solvents for assay dilution: Aqueous buffers (e.g., PBS, HEPES) with 0.1–1% DMSO v/v are commonly used; surfactants (0.01–0.05% Tween-20) can mitigate adsorption to plastics (general guidance).
  • Alternative organic solvents: DMF and NMP dissolve challenging solids but may be less assay-friendly; ethanol or methanol can work for some structures but may impact cells/enzymes at lower thresholds than DMSO.

• Practical tips

  • Prepare concentrated DMSO stocks (e.g., 10–50 mM) and confirm homogeneity by inspection and, where critical, UV/Vis or HPLC.
  • Filter through 0.22 μm PTFE if particles persist and compatibility is confirmed.
  • Avoid prolonged exposure to aqueous buffers if hydrolytic liability is unknown; prepare fresh working solutions immediately prior to use.

Storage and Reconstitution

• Item-specific storage and shipping

  • Storage Conditions: Store at -20°C (from Product Data).
  • Shipped In: Ice chest + Ice pads (from Product Data).

• General reconstitution guidance (not item-specific)

  • Allow vial to equilibrate to room temperature in a desiccator before opening to avoid moisture condensation.
  • For solid material: Dissolve in anhydrous DMSO to make a concentrated stock (e.g., 10–50 mM). If a specific solvent or concentration is required, refer to the CoA/Spec Sheet.
  • For solution format: Vortex gently; if precipitation is observed, warm mildly (≤40 °C) and sonicate briefly if needed.

• Aliquoting and stability (general)

  • Aliquot DMSO stocks into amber vials or low-bind tubes to minimize freeze–thaw cycles and adsorption.
  • Store aliquots at -20°C to -80°C. Avoid repeated freeze–thaw; thaw on ice and return promptly after use.
  • Aqueous working solutions should be prepared fresh and used promptly; discard remaining diluted solutions after the experiment unless stability has been validated.

• Specifications not provided

  • Appearance, purity, and exact solubility limits: Not specified for this item; refer to CoA/Spec Sheet.

Structure and Identity

• Item-specific (from Product Data)

  • Product Name: TrkA-IN-4 (SKU: T658654)
  • CAS: Not specified for this item; refer to CoA/Spec Sheet.
  • 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 (general/literature context for TrkA inhibitors, not item-specific)

  • TrkA inhibitors commonly feature heteroaromatic cores (e.g., pyridines, quinazolines, indazoles) that form hinge-binding H-bonds within the kinase ATP site.
  • Typical motifs include one or two H-bond donors/acceptors arranged to engage the hinge region and a hydrophobic substituent projecting into the selectivity pocket.
  • Many members are moderately lipophilic small molecules (often cLogP 2–5) with limited ionizable groups to favor cell permeability.

• 2D structure description

  • Not available for this specific item. Without a provided SMILES/InChI, a definitive 2D/functional-group description cannot be given. Please consult the CoA/Spec Sheet or SDS for definitive identity attributes.

Synthetic Utility

• This product is a target-specific small-molecule inhibitor candidate and is not typically employed as a reagent or building block in organic synthesis.

• General comments (for context only)

  • Many kinase inhibitors contain heteroaromatic scaffolds amenable to late-stage diversification (e.g., palladium-catalyzed cross-couplings on aryl/heteroaryl halides, nucleophilic aromatic substitution on activated rings). However, without the structure of TrkA-IN-4, no specific transformations can be recommended.
  • If analog synthesis is pursued, typical routes include assembling the hinge-binding core followed by selective installation of solubilizing appendages and fine-tuning lipophilicity.

• Conclusion

  • Treat TrkA-IN-4 as a finished probe molecule rather than a synthetic intermediate unless its detailed structure is available and a retrosynthetic plan has been validated.

Target Specificity

• Item-specific data are not provided for this product.

  • Target: Designation suggests inhibition of TrkA (NTRK1), but potency, selectivity versus TrkB/TrkC and broader kinome, and mode of inhibition are not specified for this item; refer to CoA/Spec Sheet and primary literature.

• General guidance for characterizing specificity (not item-specific)

  • Perform kinase panel profiling to quantify off-target interactions.
  • Use competition assays varying ATP to determine ATP-competitive behavior.
  • Apply CETSA or NanoBRET target engagement assays in cells to confirm on-target action.