AChE-IN-44 , CAS No.2758685-58-2

CAS: 2758685-58-2 Cat. No.: A1445231 Formula: C₃₁H₃₈ClN₃OS₂ Molecular Weight: 568.24

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Overview

AChE-IN-44 (Compound Tap4) is an AChE inhibitor. AChE-IN-44 can be converted into the thiazole salt AChE inhibitor Tat2.

Specifications

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Action Type

INHIBITOR

Names and Identifiers

• Molecular Weight: 568.24

Application Protocols

No vendor-validated protocols are provided for this item. The following is general literature guidance for a standard AChE inhibition assay (Ellman method); adapt and validate locally.

Ellman AChE inhibition assay (general)

• Prepare 50 mM phosphate buffer, pH 7.8, optionally with 0.01% Triton X-100.
• Make a 10–50 mM inhibitor stock in DMSO. Dilute into buffer to desired concentrations, keeping final DMSO ≤1% v/v.
• In a 96-well plate, combine per well: buffer, inhibitor (or vehicle), AChE (e.g., 0.02–0.2 U/mL; enzyme source dependent), DTNB (0.2–0.5 mM), and ATCh (0.2–1.0 mM) to start reaction.
• Read absorbance at 412 nm every 30–60 s for 5–10 min at 25°C. Determine initial rates from the linear region.
• Calculate % inhibition relative to vehicle control and fit concentration–response to obtain IC50.

Controls and checks

• Include a known AChE inhibitor as a positive control and test for detergent sensitivity (to flag aggregation-based inhibition).
• Verify that the compound does not absorb strongly at 412 nm at working concentrations (run compound + DTNB without enzyme/substrate as a blank).

Biological Roles

Biological context (general literature; not item-specific)

• Acetylcholinesterase (AChE) rapidly hydrolyzes the neurotransmitter acetylcholine into choline and acetate, terminating cholinergic signaling at synapses and neuromuscular junctions.
• AChE features a catalytic triad (Ser–His–Glu) and a deep aromatic gorge with catalytic and peripheral anionic sites. Inhibitors often exploit cation–π and hydrogen-bonding interactions within these regions.
• Inhibition of AChE is a standard method to study cholinergic pathway dynamics, synaptic transmission, and cholinesterase structure–function relationships in biochemical systems, tissues, or lysates (research use only).
• Selectivity versus butyrylcholinesterase (BChE) is biologically relevant; profiling both enzymes informs on off-target cholinesterase engagement.

Note: No item-specific potency, selectivity, or kinetic mechanism data are provided for AChE-IN-44 in this listing. Refer to the primary literature and the CoA/Spec Sheet if available.

Buffer Applications

This product is not a buffering agent. However, buffers are central to AChE enzymatic assays where this inhibitor might be used.

Common assay buffers (literature; general guidance)

• Phosphate buffer: 50 mM sodium phosphate, pH 7.4–8.0; optional 0.01% nonionic detergent to reduce nonspecific adsorption and aggregation.
• HEPES buffer: 50 mM HEPES, pH 7.4; good pH stability near physiological conditions.
• Ionic strength: Include 100–150 mM NaCl if required to mimic physiological salinity.
• Cofactors: Not required for AChE catalysis; ensure fresh preparation of chromogenic/fluorogenic substrates (e.g., ATCh and DTNB for Ellman assays).

Practical tips

• Maintain constant DMSO percentage across all wells when introducing inhibitor stocks.
• Filter buffers (0.22 µm) to reduce background scatter in optical assays and store chilled to extend substrate stability.

Green Alternatives

While the compound itself is fixed, solvent and workflow choices can improve the environmental profile of assays and handling.

Greener choices (general guidance)

• Prefer water-based assay buffers with minimal DMSO (≤1% v/v) consistent with solubility limits.
• Consider 2-MeTHF or Cyrene only for synthetic operations (if derivatization is undertaken), not for aqueous enzymology.
• Use solid dispensing (acoustic or powder pin tools) to minimize solvent use in HTS.

Comparison of common stock solvents (general)

| Solvent | Greenness notes | Assay suitability | | --- | --- | --- | | DMSO | Widely available, recyclable; moderate EHS profile | Excellent solubility; keep ≤1–2% v/v final | | Ethanol | Renewable routes possible; VOC | Variable enzyme tolerance; evaporation risk | | DMF | Problematic from EHS perspective | Avoid in biology unless indispensable |

Operational improvements

• Implement microscale assay volumes (≤50–100 µL) to reduce waste.
• Consolidate wash/conditioning steps and use multichannel/automated dispensing to minimize solvent overuse.
• Plan aliquoting to reduce freeze–thaw cycles, limiting disposal of degraded stocks.

Pharmaceutical Uses

No pharmacopeial or excipient roles are specified for this item. It is supplied strictly for research use.

• Formulation role: Not specified for this item; refer to CoA/Spec Sheet.
• Regulatory status: Not a drug product. No USP/EP monograph applicability provided.

Research/manufacturing context (general)

• May serve as a reference material or positive control in in vitro cholinesterase inhibition assays for discovery and screening workflows (non-clinical).
• For any GLP/GMP-adjacent work, qualify each lot with in-house identity/purity testing and document storage/handling history.

Physical Properties

Item-specific physical constants are not provided in the current listing.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Melting/Boiling Point: Not specified for this item; refer to CoA/Spec Sheet.
• Density/Refractive Index: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Not specified for this item; refer to CoA/Spec Sheet.
• logP/logD, pKa: Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature/practice; not item-specific)

• Many AChE inhibitors are moderately lipophilic and are typically prepared as DMSO stock solutions (e.g., 10–50 mM) for biochemical assays. If aqueous work is needed, cosolvent systems (DMSO 1–2% v/v in buffer) or inclusion of solubilizers (e.g., 0.01–0.05% nonionic surfactant) are commonly used.
• If physical data are required, determine experimentally: DSC for melting transitions, Karl Fischer for water content, shake-flask or HPLC-based logD at pH 7.4, and potentiometric/UV titration for pKa.
• Solubility screening approach: begin with DMSO, then test serial aqueous dilution into assay buffer (maintain final DMSO ≤1–2% v/v to minimize enzyme perturbation), monitoring for precipitation by light scattering or plate-reader absorbance (600–700 nm baseline). Document any colloidal behavior.

Quality & Grades

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

Interpretation and guidance

• In the absence of an explicit grade, users should consult the Certificate of Analysis (CoA) for assay purity (e.g., HPLC area %), residual solvent profile, and identity confirmation (NMR/HRMS). For screening campaigns, ≥95% HPLC purity is commonly targeted (literature best practice), but acceptability depends on application.
• For biochemical assays, low UV background and absence of reactive impurities are important. Review chromatograms for secondary peaks that could confound enzymology.
• If a stabilizer is present or recommended (e.g., acid inhibitor for base-labile moieties), it will be listed on the CoA/SDS. None are specified here.
• Batch-to-batch reproducibility: Retain aliquots of each lot used in hits-to-leads or SAR work to ensure traceability.
• If LC–MS reference data are required, request them from technical support or generate in-house and attach to your project’s compound registry.

Reaction & Applications

This product is intended as a research tool compound, particularly for probing acetylcholinesterase activity in biochemical and cellular assays.

Applications (research use; literature/general)

• AChE activity modulation: Used to inhibit AChE in Ellman-type colorimetric assays (DTNB/ATCh system) to quantify enzyme kinetics and inhibitor potency (IC50/Ki determination).
• Selectivity profiling: Comparative assays vs. butyrylcholinesterase (BChE) to determine enzyme family selectivity.
• Mechanistic studies: Time-dependent inhibition assessment to discern reversible competitive, mixed, or pseudo-irreversible (e.g., carbamylating) behavior.
• Tool in pathway studies: Modulation of cholinergic signaling in ex vivo models or lysates to study acetylcholine turnover (non-clinical research only).

Practical tips (general)

• Maintain consistent DMSO content across wells/conditions (e.g., 1% v/v) to avoid solvent effects on AChE.
• Preincubate enzyme with inhibitor (e.g., 5–15 min at room temperature) when assessing slow-binding or time-dependent inhibition.
• Confirm absence of colloidal aggregation (a known false-positive mechanism) by adding a small amount of nonionic detergent (e.g., 0.01% Triton X-100) and/or using dynamic light scattering.
• Orthogonal readouts: Complement Ellman assays with fluorogenic substrates (e.g., Amplex Red/Choline oxidase cascade) to rule out optical interference.

Reaction Conditions

No chemical reaction conditions are specified for this item. The compound is intended for use as a biochemical tool in enzyme assays rather than as a reagent.

Assay reaction conditions (literature; general guidance for AChE)

• Temperature: Ambient (22–25°C) is standard; enzyme activity is temperature dependent—maintain constant conditions across plates.
• pH: 7.4–8.0 (phosphate or HEPES buffers). Avoid extremes that denature AChE or alter inhibitor ionization.
• Substrate (Ellman): 0.2–1.0 mM acetylthiocholine (ATCh) with 0.2–0.5 mM DTNB; measure increase at 412 nm.
• Inhibitor incubation: 5–15 min preincubation with AChE may be used to assess time-dependent effects.
• Controls: Enzyme-only, substrate-only, solvent control (same DMSO %), and a known AChE inhibitor as a positive control.

Note: These are literature-level guidelines and not specifications for AChE-IN-44. Optimize for your specific enzyme source (electric eel, human recombinant, etc.).

Safety & Handling

Item-specific hazard classifications are not supplied in this listing. Always consult the SDS before use.

• GHS Classification / Pictograms / Signal Word / H-Statements: Not specified for this item; refer to SDS.
• Primary hazards (general for bioactive small molecules): May cause irritation to eyes/skin; avoid inhalation/ingestion; unknown toxicological profile—handle as a substance of unknown hazard.
• Recommended PPE: Lab coat, safety glasses, and suitable gloves (e.g., disposable nitrile). Work in a fume hood when weighing powders or preparing solutions.
• Handling tips:
  • Minimize exposure and avoid aerosolization. Use antistatic spatulas for microgram-to-milligram handling.
  • For DMSO solutions, prevent skin contact—DMSO can facilitate dermal absorption of solutes.
• Incompatibilities (general): Strong oxidizers and strong acids/bases may compromise stability for many heteroaromatic or carbamate-containing inhibitors. Do not heat above necessary temperatures.
• First aid (overview; defer to SDS): Rinse with water for skin/eye contact; move to fresh air if inhaled; seek medical attention if adverse effects occur.
• Waste disposal: Collect solutions and contaminated materials as organic hazardous waste according to institutional and regulatory guidance.

Solvent Selection

Given the lack of item-specific solubility data, select solvents empirically with biochemical assay compatibility in mind.

• Primary stock solvent (practice): DMSO is the default for many AChE inhibitors due to broad solubilizing power and miscibility with aqueous buffers at ≤1–2% v/v final.
• Secondary options (literature/general): DMF, ethanol, or 2-propanol can dissolve some scaffolds but may be less desirable in enzymatic assays; test enzyme tolerance if used.
• Aqueous work: If aqueous solutions are required, titrate compound into phosphate or HEPES buffer containing 0.01–0.05% nonionic surfactant (e.g., Tween-20 or Triton X-100) to mitigate aggregation/precipitation. Maintain consistent cosolvent across controls.
• Polarity context: AChE inhibitors span logD values; anticipate increased solubility in polar aprotic media (DMSO/DMF) relative to neat water.

Quick comparison (general, not item-specific)

• DMSO: maximal solubility; excellent for cryostorage; enzyme-compatible ≤1–2%.
• Ethanol: volatile; enzyme tolerance varies; risk of evaporation skewing concentrations.
• Aqueous buffer only: lowest solubility risk; suitable if the compound is sufficiently polar or salt-form is available.

Tip: Filter stocks through low-binding membranes (PTFE/nylon for organic, PVDF for aqueous) only if the compound is not adsorptive; otherwise, rely on visual inspection and replicate dispensing to ensure homogeneity.

Storage & Reconstitution

• Storage Conditions (item-specific): Store at −20°C.
• Shipping (item-specific): Shipped in an ice chest with ice pads.

Reconstitution and handling (general guidance; not item-specific specifications)

• Allow vial to equilibrate to room temperature in a desiccated environment before opening to avoid moisture condensation.
• Prepare concentrated DMSO stocks (e.g., 10–50 mM) for convenience. Mix thoroughly (vortex/brief sonication) until fully dissolved.
• Aliquot stock solutions into single-use portions to minimize freeze–thaw cycles. Store aliquots at −20°C in tightly sealed, inert vials.
• Protect from prolonged exposure to light and ambient humidity. If available, purge headspace with inert gas before returning to storage.
• Inspect solutions periodically for precipitation, discoloration, or apparent degradation; discard if changes are observed.

Shelf-life and stability

• Item-specific stability, hygroscopicity, and solution lifetime: Not specified for this item; refer to CoA/Spec Sheet.

Research use note

• For research use only. Not for diagnostic or therapeutic use.

Structure & Identity

AChE-IN-44 is listed as a small-molecule acetylcholinesterase (AChE) inhibitor tool compound.

• Product Name: AChE-IN-44 (research chemical)
• SKU: A1445231
• CAS: 2758685-58-2
• Category: Small molecules and compound libraries (研究/小分子)
• 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.
• CID: Not specified for this item; refer to CoA/Spec Sheet.

Structural features

• Item-specific 2D/3D structure: Not specified for this item; refer to CoA/Spec Sheet.
• General note (literature): AChE inhibitors commonly include carbamates, aryl/heteroaryl scaffolds, and cationic centers that interact with the peripheral or catalytic anionic sites of AChE. This is general background and not specific to AChE-IN-44.

Identity confirmation (guidance)

• For definitive identity, consult the product CoA/SDS for spectral data (e.g., 1H/13C NMR, HRMS) and match to the CAS registry number above.
• Consider orthogonal checks (HPLC/UPLC retention time, MS adduct patterns) when qualifying for screening libraries.

Synthetic Utility

As a bioactive small molecule tool, AChE-IN-44 is typically used as a final test article rather than as a synthetic intermediate.

• Item-specific functional groups and reactivity: Not specified for this item; refer to CoA/Spec Sheet.
• General note (literature): Scaffolds used in cholinesterase inhibition often contain aryl/heteroaryl systems, tertiary amines or quaternary centers, and in some cases carbamate or urea linkages. Such motifs can permit late-stage diversification (e.g., amide coupling, N-alkylation, Suzuki-type arylation) for SAR exploration. This is general information and not specific to AChE-IN-44.

If derivatization is pursued

• Establish a baseline potency profile before modifications.
• Retain a synthetic route that tolerates salt formation for improved handling/solubility if needed (e.g., HCl or mesylate salts), subject to compound stability.
• Monitor lipophilicity and polar surface area to balance AChE binding with assay solubility.

Target Specificity

• Item-specific target data (e.g., Ki/IC50 values, selectivity vs. BChE or other serine hydrolases) are not specified for this item; refer to CoA/Spec Sheet.

General guidance

• When deploying as an AChE tool compound, determine potency against the specific AChE isoform/species used in your assay, and profile against BChE to quantify selectivity.
• Use orthogonal assay formats (colorimetric, fluorogenic, MS-based activity assays) to confirm on-target effects and exclude assay interference.