AChE-IN-25 , CAS No.2835538-69-5

CAS: 2835538-69-5 Cat. No.: A1445170 Formula: C₂₀H₁₅ClN₄O₄S Molecular Weight: 442.88

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Overview

AChE-IN-25 is a potent, selective and uncompetitive acetylcholinesterase ( AChE ) inhibitor ( IC 50 =2.95 µM). AChE-IN-25 can be used in Alzheimer's disease research.

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

442.88

Application Protocols

Item-specific validated applications and recommended conditions

• Not specified for this item; refer to CoA/Spec Sheet and SDS. No in-house validated protocols, dilutions, or positive-control conditions are provided for AChE-IN-25.

Guidance for developing your own protocol (general, not item-specific)

• Stock preparation: Prepare a concentrated DMSO solution (literature practice: 10–50 mM) and aliquot to avoid freeze–thaw. Verify solubility visually and by LC–MS if available.
• Dose–response: Run ≥10-point serial dilutions covering at least 3–4 logs of concentration with constant cosolvent across the plate. Include vehicle and reference inhibitor controls (if available in your lab).
• Assay selection: Ellman’s colorimetric assay or fluorescence-based substrates are commonly used for AChE. Validate dynamic range, Z′ factor, and solvent tolerance before inhibitor testing.
• Data analysis: Fit to a four-parameter logistic model for IC50; for mechanism and Ki, conduct initial-rate measurements at multiple substrate concentrations and analyze with appropriate kinetic models (e.g., competitive vs. mixed).
• Counterscreens: Test against BChE and include aggregation controls (e.g., 0.01–0.05% nonionic detergent) to identify nonspecific inhibition.

Note

• The above are generic best practices from the literature. They are not validated for this specific item. Always adapt to your enzyme source, substrate, and instrumentation.

Biological Roles

General background (biology of the intended target; not item-specific)

• Acetylcholinesterase (AChE, EC 3.1.1.7) is a serine hydrolase responsible for rapid hydrolysis of the neurotransmitter acetylcholine into choline and acetate, thereby terminating cholinergic signaling at synapses and neuromuscular junctions.
• AChE features a deep active-site gorge containing the catalytic triad (Ser–His–Glu), an oxyanion hole, a choline-binding site, and a peripheral anionic site that can modulate substrate access and bind allosteric ligands. Inhibitors may target the catalytic site, the peripheral site, or bridge both, leading to distinct kinetic behaviors (competitive, mixed, or noncompetitive inhibition).
• In model systems, AChE activity impacts synaptic transmission, learning paradigms, and motor function. Experimental modulation with inhibitors is widely used to probe cholinergic pathways, structure–function relationships, and enzymatic mechanisms.
• Selectivity considerations: Butyrylcholinesterase (BChE) shares structural homology with AChE but differs in substrate specificity and gorge architecture; profiling against BChE is common to establish target preference.
• Assay environment: Ionic strength, pH (typically near neutral), and presence of detergents can influence enzyme conformation and inhibitor binding. DMSO fractions should be kept low and consistent across wells to avoid artifacts.

Important

• The above describes general biology. AChE-IN-25’s actual potency, mode of inhibition, and selectivity are not specified for this item; users must determine these experimentally with their assay systems.

Buffer Applications

Relevance

• AChE-IN-25 is not a buffering agent and is not used to maintain pH directly. However, appropriate buffer systems are essential for reliable acetylcholinesterase assays.

General assay buffer guidance (literature; not item-specific)

• Common buffers: Phosphate (50–100 mM, pH 7.2–7.6) or HEPES (25–50 mM, pH 7.2–7.6) are frequently used for AChE activity assays. Maintain constant ionic strength.
• Additives: 0.01–0.05% nonionic detergent (e.g., Tween-20) can reduce nonspecific adsorption of hydrophobic inhibitors. Include 0.1% BSA or similar carrier if stickiness is observed. Avoid chelators or reductants unless validated for your enzyme preparation.
• Temperature: Perform assays at a controlled temperature (e.g., 25–37°C) and equilibrate plates to minimize drift.
• Solvent control: Keep the final DMSO (or other cosolvent) fraction constant across all wells, typically ≤1% v/v, to minimize solvent effects on enzyme activity.

Item-specific data

• Buffer compatibility or preferred compositions for AChE-IN-25: Not specified for this item; refer to CoA/Spec Sheet. Optimize empirically for your enzyme source (human recombinant vs. tissue extract) and substrate system.

Green Alternatives

Context

• As a small-molecule inhibitor used mainly in biochemical assays, the principal green-chemistry leverage points are solvent choice, waste minimization, and plate-based microvolumes rather than changes to the compound itself.

Greener solvent practices (general guidance)

• Favor water-based buffers with minimal organic cosolvent (≤1% v/v) once the compound is dissolved.
• Prefer DMSO over DMF/NMP when possible due to lower reproductive toxicity concerns and better handling practices, though DMSO still requires care because it enhances dermal absorption.
• For analytics, choose acetonitrile judiciously versus methanol, balancing toxicity, waste, and chromatographic performance. Recycle solvents where facility infrastructure allows.

Operational strategies

• Miniaturize: Use 384- or 1536-well formats to cut solvent consumption and chemical waste.
• Right-first-time: Conduct small pilot solubility and stability checks to avoid failed large-batch preparations.
• Closed systems: Employ sealed plates and low-evaporation lids to reduce emissions of volatile organics and maintain concentration fidelity.

Comparison snapshot (general)

• DMF/NMP: Powerful but undesirable from a green perspective; avoid unless essential.
• DMSO: Broadly accepted compromise; miscible with water and widely compatible with enzyme assays.
• Ethanol/IPA: More benign but lower solvency; suitable for less hydrophobic compounds.

Note: No item-specific solvent or stability data are provided; confirm compatibility empirically and consult institutional green-chemistry guidelines.

Pharmaceutical Uses

Scope and limitations

• This product is supplied strictly for research use only and is not intended for human or veterinary use, drug substance manufacturing, or clinical diagnostics.

Formulation and development contexts (general; not item-specific)

• Tool compounds such as AChE-IN-25 may be used in preclinical research workflows as reference inhibitors in biochemical assays, ex vivo systems, or as positive controls in screening cascades. In formulation research, small-scale solubility, stability, and salt-screen studies can inform handling but do not imply suitability for clinical development.
• Excipients and compendial status: No pharmacopeial monographs or excipient roles are indicated for this item. If the compound were to be used as an analytical reference, documentation would include identity confirmation and purity, typically provided through a CoA.

Item-specific status

• Pharmacopeia listing, GMP grade, or excipient designation: Not specified for this item; refer to CoA/Spec Sheet.

Compliance note

• Any scale-up, GLP/GMP use, or incorporation into regulated workflows requires independent qualification, method validation, and risk assessment beyond the scope of this research-use product.

Physical Properties

Item-specific properties (this lot)

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

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

• Physical state: Many screening compounds are crystalline solids or amorphous powders supplied in vials or plated formats; they are often formulated as DMSO stock solutions for biochemical assays.
• Solubility practice: Anhydrous DMSO is the most common primary solvent for preparing concentrated stocks (e.g., 10–50 mM, literature practice), followed by dilution into aqueous assay buffers containing surfactant (e.g., 0.01–0.05% nonionic detergent) to mitigate precipitation and nonspecific binding.
• Ionization: If the compound is basic, salts (e.g., HCl) may increase aqueous solubility; if acidic, sodium/potassium salts are sometimes used. Absent a structure, do not assume ionization behavior.
• Stability considerations: Many drug-like molecules are light- and moisture-sensitive to varying degrees; minimize exposure to light, air, and elevated temperature, and avoid repeated freeze–thaw of stock solutions by aliquoting.

Important: For quantitative parameters and solvent compatibility of AChE-IN-25 (CAS 2835538-69-5), defer to the product’s CoA/Spec Sheet and SDS.

Quality & Grades

Item-specific quality information

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

Interpretation and expectations (general guidance)

• Research-use designation: This product is supplied strictly for research and laboratory use (not for diagnostic, therapeutic, or clinical applications). In this context, quality is typically assured via identity confirmation (e.g., NMR/LC–MS, HRMS) and purity determination (e.g., HPLC area %).
• What “purity” means: Reported purity, when available on the CoA, commonly reflects HPLC/UPLC area percent at a specified wavelength. Trace metals, water content (Karl Fischer), residual solvents, and salt content may or may not be specified.
• What to check on receipt: Verify lot-specific identity (spectral data), purity, appearance, and recommended storage. Confirm any stabilizers or salt form (free base vs. HCl, etc.) that could influence solubility and assay setup.
• UV/LC considerations: If HPLC-grade qualities (low UV baseline bleed) are important for your analytics, confirm UV cutoff and baseline behavior for your mobile phase. Absent such data, pre-run a blank and standard to ensure clean baselines.

Documentation

• For definitive specifications, chromatographic purity, residual solvent profiles, water content, metal limits, and any stabilizer information, consult the lot-specific CoA/Spec Sheet. If not present, contact technical support to request analytical documentation.

Reaction & Applications

Scope of use (research only)

• Functional role: AChE-IN-25 is intended as a research tool compound for probing acetylcholinesterase (AChE) activity and inhibition in biochemical assays, target validation studies, and selectivity profiling panels. No medical/clinical use is intended or implied.

Enzymology and assay applications (general guidance; not item-specific)

• Primary assay formats: Colorimetric Ellman assay (DTNB/ATCh), fluorometric substrates (e.g., Amplex-type readouts), and coupled enzyme systems. These measure AChE catalytic turnover in the presence of inhibitor to derive IC50 and, with multiple substrate concentrations, mechanism (competitive, noncompetitive, mixed) and Ki values.
• Selectivity: Parallel testing against BChE (butyrylcholinesterase) and other serine hydrolases helps delineate target preference and off-target risk.
• Kinetic considerations: Maintain substrate concentrations around Km and include time-course sampling to verify steady-state. Use minimal and constant cosolvent, and preincubate inhibitor with enzyme where slow-binding behavior is suspected.
• Orthogonal confirmation: Use LC–MS to confirm chemical stability of the inhibitor under assay conditions. Thermal shift assays (DSF) or SPR/ITC can provide binding evidence independent of catalysis.

Chemical reactions

• This item is not primarily used as a synthetic reagent. If derivatization or immobilization is desired for probe development, obtain structural data and plan modifications away from pharmacophore features.

Note: Because no structural or potency data are provided here, users should perform pilot solubility checks and preliminary dose–response curves before large-scale studies.

Reaction Conditions

Not typically applicable

• AChE-IN-25 is not provided as a reagent for performing chemical transformations; therefore, there are no canonical “reaction conditions” to report.

Assay conditions (general, literature-based; not item-specific)

• If using this compound in enzymology experiments, representative assay conditions for AChE include: neutral pH buffers (e.g., phosphate or HEPES, pH ~7.4), controlled temperature (25–37°C), low and constant cosolvent (≤1% DMSO), and appropriate substrate concentrations near Km to determine mechanism and Ki.
• Preincubation: Where slow-binding or time-dependent inhibition is suspected, preincubate enzyme with inhibitor (e.g., 5–30 min) before initiating the reaction.

Item-specific data

• Optimal temperatures, incubation times, and concentrations for AChE-IN-25: Not specified for this item; refer to CoA/Spec Sheet and determine empirically in your assay system.

Safety & Handling

Item-specific hazard data

• Signal word: Not specified for this item; refer to SDS.
• H-statements and GHS classification/pictograms: Not specified for this item; refer to SDS.

General laboratory precautions (professional guidance; not item-specific)

• Treat as a potentially hazardous small molecule. Many acetylcholinesterase inhibitors can present significant toxicity risks via inhalation, dermal exposure, or ingestion. Handle in a certified chemical fume hood with appropriate containment.
• Recommended PPE: Lab coat, safety glasses or chemical splash goggles, and disposable nitrile gloves (double-gloving advisable for concentrated solutions). Change gloves frequently; avoid skin contact.
• Handling: Avoid aerosol generation. Use dedicated tools for weighing and solution prep. When preparing DMSO stocks, prevent contact with skin, as DMSO can enhance dermal absorption of solutes.
• Incompatibilities: Absent structural data, avoid strong oxidizers and strong acids/bases until compatibility is confirmed. Keep away from ignition sources if flammable solvents are in use.
• First aid overview (defer to SDS for authoritative instructions): In case of skin contact, wash thoroughly with soap and water. For eye contact, rinse cautiously with water for several minutes and seek medical attention. If inhaled, move to fresh air and monitor for symptoms; seek medical attention. If swallowed, rinse mouth; do not induce vomiting unless directed by medical personnel.
• Spill/accident response: Evacuate area as needed, ventilate, and use inert absorbent for liquids. Dispose in accordance with institutional and local regulations.

Always consult the product-specific SDS for definitive hazard classification and response guidance.

Solvent Selection

Applicability

• AChE-IN-25 is supplied as a small-molecule tool compound. Solvent choice primarily concerns stock-solution preparation for biochemical/biophysical assays and, secondarily, analytical characterization.

Item-specific solvent data

• Solubility and preferred solvents: Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature/typical practice)

• Primary solvent: Anhydrous DMSO is the standard for preparing concentrated stocks (e.g., 10–50 mM, literature practice) thanks to its broad solubilizing power and miscibility with aqueous buffers.
• Secondary solvents: Ethanol or DMF may be used for less DMSO-tolerant systems; however, DMF is less desirable from a safety and green-chemistry perspective. Where possible, transition to aqueous buffers with small amounts of cosolvent (≤1–2% v/v) after initial dissolution.
• Aqueous work: Once dissolved in DMSO or ethanol, gradual dilution into assay buffer (e.g., phosphate or HEPES with 0.01–0.05% nonionic surfactant) can reduce precipitation and nonspecific adsorption.
• Analytics: For LC–MS, water/acetonitrile (0.1% formic acid) or water/methanol systems are typical. Verify ionization and retention behavior empirically.

Comparison (general)

• DMSO: Highest solvency; widely accepted in enzyme assays up to 0.5–2% v/v.
• Ethanol: Lower solvency; broader biological tolerance in some systems.
• 2-Propanol/Acetonitrile: Useful in analytics; less common for biological stocks; can denature proteins at modest percentages.

Always confirm assay tolerance to the chosen cosolvent and keep the final solvent fraction minimal and constant across controls.

Storage & Reconstitution

Item-specific instructions

• Storage Conditions: Store at -20°C.
• Shipped In: Ice chest + Ice pads.

Additional handling guidance (general; not item-specific)

• Protect from light and moisture. Upon receipt, allow the sealed container to equilibrate to room temperature before opening to minimize moisture condensation.
• Reconstitution: Specific solvent and concentration recommendations are not provided for this item; refer to CoA/Spec Sheet. Literature practice for small-molecule inhibitors is to prepare concentrated stocks in anhydrous DMSO (e.g., 10–50 mM), then aliquot and store at -20°C or -80°C.
• Freeze–thaw: Avoid repeated freeze–thaw cycles by preparing single-use aliquots. Thaw on ice and mix gently before use; do not vortex if foaming is a concern.
• Aqueous stability: Unknown for this item. Prepare working dilutions fresh in assay buffer, keep on ice or at controlled room temperature during use, and discard remaining diluted solutions after the experiment unless stability has been verified.
• Container considerations: Use low-bind tubes and solvent-resistant caps. For plate-based work, use low-evaporation sealing films.

Compliance note

• For any specific stability (t90), peroxide/water content, or re-test date, consult the lot-specific CoA/Spec Sheet. When in doubt, perform a quick-check by LC–MS/UPLC to confirm integrity prior to critical assays.

Structure & Identity

• SKU: A1445170
• Product Name: AChE-IN-25
• CAS: 2835538-69-5
• Category: Small molecule tool compound (putative acetylcholinesterase inhibitor; inference from name only)

Item-specific identifiers

• 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

• Functional groups, ring systems, and stereochemistry: Not available for this item (no structure provided). Without a structure, a definitive description of heteroatoms, aromaticity, H-bond donors/acceptors, or chiral centers cannot be given.

Context and scope (general)

• Naming convention: “AChE-IN-25” denotes a numbered entry in an acetylcholinesterase inhibitor set or screening collection. This is a naming heuristic and not a verification of identity or selectivity.
• If a structure becomes available, chemists typically document: heteroatom count, rotatable bonds, polar surface area, cLogP/cLogS, pKa(s), and likely ionization state near physiological pH to anticipate solubility, permeability, and assay behavior.

2D structure (descriptive)

• Not possible to depict or describe without the molecular drawing. Please consult the product’s CoA/Spec Sheet or contact technical support for the structural record associated with CAS 2835538-69-5.

Synthetic Utility

Applicability

• AChE-IN-25 is supplied as a finished small-molecule tool compound intended for biochemical studies, not as a synthetic intermediate or reagent.

General commentary (not item-specific)

• Without a disclosed structure, no guidance can be offered on functional group manipulations, protecting-group strategies, or diversification routes. If analog generation or probe attachment (e.g., biotinylation, fluorophore tagging) is desired, obtain structural information and SAR to identify modification-tolerant positions.
• For analytical derivatization (e.g., forming a reporter conjugate), typical strategies include installing linkers through noncritical moieties distant from the pharmacophore; however, this requires prior knowledge of binding determinants.

Conclusion

• Synthetic applications are not typically relevant for this item. Users interested in chemical modifications should consult the structure (when available) and plan bespoke syntheses with due consideration for IP, safety, and scale.

Target Specificity

Item-specific data

• Verified primary target, isoform selectivity, species reactivity, and off-target profile: Not specified for this item; refer to CoA/Spec Sheet or associated technical notes.

General considerations (not item-specific)

• Nomenclature suggests intended activity at acetylcholinesterase (AChE), but without potency or selectivity data, no claims can be made. Establish specificity experimentally by:
  • Profiling against human AChE versus BChE and other serine hydrolases.
  • Testing multiple species sources if cross-species comparisons are needed.
  • Employing orthogonal binding/biophysical assays (e.g., SPR/ITC/DSF) and activity-based protein profiling where appropriate.
• Watch for assay artifacts due to aggregation, redox cycling, or fluorescence interference; include detergent controls and counter-screens.

Conclusion

• No target-specific guarantees are provided in this listing. Users should generate their own target-engagement and selectivity data for AChE-IN-25 under their experimental conditions.