Hesperadin - Moligand™, 10mM in DMSO , Inhibitor of aurora kinase B, CAS No.422513-13-1, Inhibitor of aurora kinase B

CAS: 422513-13-1 Cat. No.: H409181 分子式: C₂₉H₃₂N₄O₃S 分子量: 516.67 EC番号: 875-899-3

グレード＆純度 Moligand™, 10mM in DMSO

Synonyms

(Z)-N-(2-oxo-3-(phenyl(4-(piperidin-1-ylmethyl)phenylamino)methylene)indolin-5-yl)ethanesulfonamide

Storage

Store at -80°C

Shipped In

Dry ice packs + Cold packs

概要

Information

Hesperadin potently inhibitsAurora BwithIC50of 250 nM in a cell-free assay. It markedly reduces the activity of AMPK, Lck, MKK1, MAPKAP-K1, CHK1 and PHK while it does not inhibit MKK1 activity in vivo.
In vitro

Hesperadin inhibits the ability of immunoprecipitated Aurora B to phosphorylate histone H3 with IC50 of 250 nM and markedly reduces the activity of other kinases (AMPK, Lck, MKK1, MAPKAP-K1, CHK1, and PHK) at a concentration of 1 μM. In contrast, only 20-100 nM of Hesperadin is sufficient to induce the loss of mitotic histone H3-Ser10 phosphorylation in HeLa cells. Hesperadin treatment causes defects in mitosis and cytokinesis, leading to stoppage of proliferation of HeLa cells and polyploidization, which can be specifically ascribed to the inhibition of Aurora B function during the process of chromosome attachment. Hesperadin (100 nM) quickly overrides the mitotic arrest induced by taxol or monastrol but not by nocodazole. Hesperadin and nocodazole treatment in HeLa cells abolishes kinetochore localization of BubR1 and diminishes the intensity of Bub1 at kinetochores, suggesting that Aurora B function is required for efficient kinetochore recruitment of BubR1 and Bub1, which in turn might be necessary for prolonged checkpoint signaling. Hesperadin prevents the phosphorylation of recombinant trypanosome histone H3 by the T. brucei Aurora kinase-1 (TbAUK1) from pathogenic Trypanosoma brucei with IC50 of 40 nM in vitro kinase assays. Hesperadin significantly inhibits cell growth of cultured infectious bloodstream forms (BF) with IC50 of 48 nM, and only weakly inhibits cell growth of insect stage procyclic forms (PF) with IC50 of 550 nM.

In vivo

Cell Data

cell lines：

Concentrations：Final concentration ~500 nM

Incubation Time：24 and 48 hours

Powder Purity:≥99%

仕様

同義語

(Z)-N-(2-oxo-3-(phenyl(4-(piperidin-1-ylmethyl)phenylamino)methylene)indolin-5-yl)ethanesulfonamide

仕様と純度

Moligand™, 10mM in DMSO

生化学的・生理学的メカニズム

Hesperadin potently inhibits Aurora B with IC50 of 250 nM in a cell-free assay. It markedly reduces the activity of AMPK, Lck, MKK1, MAPKAP-K1, CHK1 and PHK while it does not inhibit MKK1 activity in vivo.

保管条件

Store at -80°C

入荷

Dry ice packs + Cold packs

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等級

Moligand™

アクションタイプ

INHIBITOR

作用メカニズム

Inhibitor of aurora kinase B

名前と識別子

異性体SMILES

CCS(=O)(=O)NC1=CC2=C(C=C1)NC(=C2C(=NC3=CC=C(C=C3)CN4CCCCC4)C5=CC=CC=C5)O

分子量

516.67

Reaxy-Rn

41774234

Reaxys-RN_link_address

https://www.reaxys.com/reaxys/secured/hopinto.do?context=S&query=IDE.XRN=41774234&ln=

関連ターゲット（人間）

AURKB Tchem Aurora kinase B (1 Activities)

Discover Target: AURKB

Activity Type	Activity Value -log(M)	Mechanism of Action	Activity Reference	Publications (PubMed IDs)

化学的性質と物理的性質

• 溶解度: Solubility (25°C) In vitro DMSO: 86 mg/mL (200.22 mM); Ethanol: 86 mg/mL (200.22 mM); Water: Insoluble
• 融点（°C）: >226°C

Application Protocols

No vendor-validated protocols are provided for this specific item. The following is general, non-item-specific guidance for deploying small-molecule ligands in research assays. Optimize for your system.

• Stock solution and aliquoting:
  • Prepare a 10–50 mM stock in anhydrous DMSO. Mix until fully dissolved. Pass through a 0.22 µm PTFE filter if particulates persist.
  • Dispense into single-use aliquots (e.g., 10–50 µL) in low-binding tubes. Store at −80°C per product instructions.
• Biochemical enzyme assay (general):
  • Prepare serial 3-fold dilutions in assay buffer to span a broad range (e.g., 0.5 nM–50 µM). Keep final DMSO constant across wells (≤1% v/v, or assay-dependent maximum).
  • Include vehicle-only controls and, if available, a known reference inhibitor for benchmarking.
  • Readout: Follow your assay’s detection modality (fluorescence, luminescence, radiometric). Fit concentration–response curves with a 4-parameter logistic model.
• Cell-based assay dosing (general):
  • Pre-dilute the DMSO stock 1:100–1:1000 into medium to achieve final DMSO ≤0.1–0.5% v/v.
  • Titrate exposure time (e.g., 1–24 h) according to the biological question and cell tolerance.
  • Monitor cytotoxicity/viability in parallel to decouple on-target effects from nonspecific toxicity.
• Recordkeeping and QC:
  • Document lot number, stock concentration verification, and storage history.
  • Inspect visually for precipitation before each use; discard compromised aliquots.

These procedures are suggestions only and should be validated in your laboratory.

Biological Roles

Scope: The following provides general, literature-level context for small-molecule ligands used to interrogate mitotic kinase pathways. It is not item-specific potency/target data. Consult primary literature and perform in-house validation for your system.

• Chemical biology role (literature/general): Compounds under the name Hesperadin are widely used as chemical probes in cell-cycle research to perturb mitotic progression and to study the roles of serine/threonine kinases associated with chromosome alignment and segregation.
• Mechanistic theme (literature/general): Many reports describe ATP-competitive binding within kinase active sites, enabling transient, reversible modulation of phosphorylation events during mitosis. The phenotypic consequences often include altered spindle checkpoint signaling and chromosome dynamics in cultured cells.
• Utility in assays: Employed in enzyme inhibition assays, cellular phospho-biomarker readouts, live-cell imaging of mitosis, and chemical genetics studies. Users typically generate dose–response curves to establish working concentrations that balance on-target effects and cell health.
• Off-target considerations: As with most kinase-directed ligands, selectivity can be context-dependent across cell types and assay conditions. Profiling against kinase panels and orthogonal confirmation (e.g., rescue with resistant mutants) are recommended practices (general guidance).

Important: No medical or therapeutic claims are made. This product is for research use only. For precise target identity, selectivity metrics, and potency values for the specific lot, consult the CoA and independently curated literature sources.

Buffer Applications

This item is not a buffering reagent. However, appropriate buffer systems are critical when deploying it in biochemical or cell-based assays.

General buffer guidance (assay context; not item-specific):

• Enzymatic assays: Use a well-defined buffer (e.g., 20–50 mM HEPES or Tris, pH 7.0–7.5) with Mg2+/Mn2+ as required by the kinase system. Maintain ionic strength with 100–150 mM NaCl and include 0.01–0.05% Tween-20 to reduce nonspecific binding if compatible.
• Cellular assays: Culture media should maintain physiological pH/CO2 conditions. When dosing from DMSO stocks, ensure final DMSO is minimized (≤0.1–0.5% v/v typically).
• Solubility aids: If precipitation occurs upon dilution, consider pre-dilution of the DMSO stock 1:10 into buffer containing protein (e.g., 0.1% BSA) or cyclodextrin, provided these components do not interfere with readouts.
• Adsorption control: Low-binding plasticware can mitigate loss of hydrophobic ligands at low nM–µM concentrations.

Always validate that buffer components do not chelate essential cofactors or interfere with detection (fluorescence/absorbance) in your specific assay.

Green Alternatives

Because this product is a bioactive small molecule primarily used as a research ligand, “green alternatives” relate to solvent and workflow choices rather than replacing the compound itself.

Greener solvent considerations for solution preparation (general):

• Replace or minimize high-boiling amide solvents (e.g., DMF) with DMSO or aqueous-compatible media when assay permits.
• Keep DMSO to the lowest effective final concentration in assays to reduce environmental load and facilitate aqueous waste handling.
• For preparative dissolution prior to bioassays, consider ethanol or aqueous buffer with solubilizers (cyclodextrins, low % nonionic surfactant) if compound solubility and assay tolerance allow.

Comparison (general guidance):

• DMSO vs DMF: DMSO is often preferred for biotesting due to lower volatility and better biocompatibility at low percentages.
• Ethanol vs isopropanol: Ethanol typically has a more favorable environmental and safety profile; however, solvency for heteroaromatic ligands may be limited.
• Aqueous buffer + cyclodextrin: Can improve apparent solubility while avoiding organic solvent in the final assay; validate for interference.

Operational practices to enhance sustainability:

• Use small, single-use aliquots to reduce waste from degraded stocks.
• Adopt miniaturized assay formats (384/1536-well) to diminish solvent consumption.
• Segregate DMSO-containing waste for proper disposal per institutional EHS guidance.

Pharmaceutical Uses

This product is supplied strictly for research use only and is not intended for human or veterinary use in drugs, diagnostics, or clinical applications.

Formulation and development context (general, non-clinical):

• As a reference ligand or tool compound, materials of this class may be used in preclinical research to benchmark assay systems, evaluate target engagement methodologies, or serve as positive controls in screening cascades.
• Typical laboratory formulations include:
  • Concentrated DMSO stock solutions (e.g., 10–50 mM) stored in aliquots at subzero temperatures.
  • For in vivo research method development (non-clinical), co-solvent systems such as PEG-400/saline or Captisol may be explored by researchers to understand exposure and PD relationships; such uses require rigorous internal safety and ethics approvals and are outside the scope of this catalog listing.

Regulatory note:

• No pharmacopeial status is claimed.
• No claims are made regarding safety, efficacy, or therapeutic utility.
• Users are responsible for ensuring compliance with institutional and regulatory requirements for any non-clinical exploratory work.

Physical Properties

Item-specific properties provided in Product Data:

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

Literature/general expectations for small-molecule screening compounds (for planning only; verify with CoA/SDS):

• Physical state: Typically supplied as a solid (free base or salt) suitable for dissolution in DMSO for assay stock solutions (literature/general).
• Solubility behavior: Many kinase-focused ligands are readily soluble in anhydrous DMSO (e.g., 10–50 mM stocks) and may require co-solvent or surfactant to solubilize in purely aqueous buffers (literature/general). Actual solubility of this item is not specified; confirm experimentally.
• LogP/pKa: Not specified for this item.
• Melting point, density, refractive index, UV-Vis characteristics: Not specified for this item; refer to CoA/Spec Sheet.

Practical notes for handling small-molecule library members (general):

• Prepare concentrated DMSO stocks under dry, ambient conditions, then aliquot to minimize freeze–thaw cycles.
• When diluting into aqueous media, keep final DMSO ≤0.1–0.5% v/v for cellular systems or as tolerated by the assay.
• If precipitation occurs upon aqueous dilution, increase dilution steps, include co-solvent (e.g., 0.01–0.1% Tween-80) where assay-compatible, or gently warm and vortex prior to use.

Always defer to the SDS for authoritative physical/chemical hazard data and the CoA for lot-specific specs.

Quality and Grades

• Item-specific grade (from Product Data): Moligand™.

What Moligand™ implies (general description of grade and intended use):

• Moligand™ denotes a research-only, small-molecule ligand suitable for discovery workflows, screening, and chemical biology assays. Materials in this grade are typically provided with identity confirmation (e.g., NMR/LC–MS) and purity appropriate for screening. Exact acceptance criteria, analytical methods, and purity thresholds are not specified here; refer to the CoA/Spec Sheet.
• Stabilizers, salt form, residual solvents, water content, metal content, and UV cutoffs are Not specified for this item; refer to CoA/Spec Sheet.

Analytical considerations for users:

• If employing the compound in quantitative biology/biophysics, obtain and review the CoA for lot-specific purity, counterion/salt information, and residual solvent profile.
• For high-sensitivity assays (e.g., biophysical binding, proteomics-enabled target engagement), consider in-house verification by orthogonal methods (LC–MS, qNMR) and document stock concentrations by absorbance or quantitative NMR where applicable.
• For SAR or reference control purposes, maintain lot traceability and record any observed polymorph/solvate behavior if encountered.

Regulatory note: This product is for research use only and is not intended for human or animal therapeutic, diagnostic, or clinical applications.

Reaction and Applications

This product is supplied as a bioactive small molecule for screening and target interrogation, not as a synthetic reagent. Consequently, traditional organic reaction applications (e.g., cross-coupling, Grignard, oxidations) are not the primary use case for this item.

Research application contexts (general, non-clinical):

• Chemical biology: Employed as a tool compound candidate in pathway dissection, cell-cycle studies, and kinase-centric assay development (literature/general; confirm target scope for your study independently).
• High-throughput screening (HTS) and profiling: Suitable for inclusion in focused libraries, dose–response studies, and orthogonal assay confirmation when handled under screening-compatible solvent conditions.
• Mechanistic cell studies: May be used to perturb mitotic processes and evaluate phenotypic outcomes in non-clinical research models when such use is supported by independent literature sources.

If you require a reagent for synthetic transformations, consider selecting from our catalog of building blocks, catalysts, and solvents designed for preparative chemistry. For this item, prioritize sections on Solvent Selection, Biological Roles, Application Protocols, and Storage.

Reaction Conditions

Not primarily applicable: This product is a bioactive small molecule intended for screening and assay use, not for performing synthetic transformations.

Assay preparation conditions (general guidance; not item-specific):

• Stock preparation: Dissolve in anhydrous DMSO to prepare 10–50 mM stocks. Sonication or gentle warming (≤37°C) may aid dissolution. Filter if necessary through 0.22 µm PTFE.
• Working solutions: Prepare serial dilutions in assay buffer immediately before use to achieve desired concentrations (commonly nM–µM ranges, determined by your assay’s sensitivity and tolerance).
• Temperature: Many biochemical assays are run at 25–30°C; cell-based assays at 37°C with 5% CO2. Protect solutions from prolonged light and avoid repeated freeze–thaw.
• Controls: Include vehicle controls matching the final DMSO percentage and, where possible, orthogonal positive/negative controls to confirm specificity.

For information on actual chemical reaction conditions (solvent, catalysts, temperatures) pertaining to the synthesis of related scaffolds, consult medicinal chemistry literature; such data are not part of this product specification.

Safety and Handling

GHS/Hazard details in Product 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 safety guidance for small-molecule bioactives (not substance-specific; follow your institution’s EHS policies):

• PPE: Lab coat, safety glasses, and appropriate chemically resistant gloves (e.g., nitrile). Use a certified chemical fume hood during weighing and solution preparation.
• Avoid inhalation of dusts/aerosols and skin contact. Prevent ingestion and environmental release. Wash thoroughly after handling.
• First aid overview (general):
  • Inhalation: Move to fresh air; seek medical attention if symptoms persist.
  • Skin/eye contact: Rinse with plenty of water for ≥15 minutes; remove contaminated clothing; seek medical attention for persistent irritation.
  • Ingestion: Rinse mouth; do not induce vomiting; seek medical advice.
• Storage incompatibilities (general): Avoid strong oxidizers and extreme pH during storage of solutions unless stability is demonstrated. Store dry and cold as directed below.
• Special risks: Bioactive screening compounds may exhibit potent cellular effects at low concentrations. Handle as potentially toxic and minimize exposure. Use dedicated, clearly labeled containers to prevent cross-contamination of screening collections.

Authoritative information (hazards, exposure limits, spill/cleanup, disposal) must be taken from the substance-specific SDS accompanying this product.

Solvent Selection

Applicability: As a bioactive small molecule for screening, solvent selection centers on reliable stock solution preparation and compatible assay dilution.

• Item-specific solubility data: Not specified for this item; verify experimentally and/or consult the CoA.

General solvent guidance for small-molecule ligands (literature/practice-based):

• Primary solvent: Anhydrous DMSO is the standard for 10–50 mM screening stocks due to high solvency and stability.
• Aqueous dilution: For biochemical assays, dilute DMSO stocks into assay buffer while keeping final DMSO ≤0.1–1.0% v/v depending on assay tolerance. For cell-based assays, ≤0.1–0.5% is commonly used to limit solvent effects (literature/general).
• Alternative co-solvents: DMF or ethanol can sometimes aid solubility, but may be less assay-compatible; evaluate carefully.
• pH considerations: If the compound is ionizable, apparent solubility may improve in buffered media at appropriate pH. Without item-specific pKa, titration screening is advised.

Comparison (general):

• DMSO: Maximal solvency, excellent for archiving; may affect some enzymes/cells at >0.5%.
• Ethanol: More biocompatible at low levels; lower solvency for many heteroaromatics.
• PEG-400: Useful for in vivo formulation research (not clinical); viscous and can interfere with some readouts.

Practical tips:

• Warm and vortex DMSO stocks if slow-dissolving; avoid high heat.
• Filter 0.22 µm if particulate persists and the compound is not filter-adsorbed.
• Prepare single-use aliquots to limit freeze–thaw cycles.

Storage and Reconstitution

Item-specific storage/shipping (from Product Data):

• Storage Conditions: Store at −80°C.
• Shipped In: Dry ice packs + cold packs.

General guidance for reconstitution and handling (not item-specific; follow CoA/SDS and internal SOPs):

• Upon receipt: Keep frozen. Allow containers to equilibrate to room temperature in a desiccator before opening to minimize moisture condensation.
• Reconstitution:
  • Solvent: Use anhydrous DMSO for initial stock preparation unless the CoA specifies otherwise. Stock concentration of 10–50 mM is commonly practical for screening workflows.
  • Technique: Add solvent incrementally, vortex, and, if necessary, sonicate briefly. Avoid prolonged heating or exposure to light.
• Aliquoting and storage:
  • Prepare single-use aliquots in chemically resistant, low-binding tubes. Store at −80°C in the dark.
  • Minimize freeze–thaw cycles. Thaw only the amount needed for immediate use.
• Stability considerations:
  • Solution stability, sensitivity to light, air, or moisture: Not specified for this item; refer to CoA/Spec Sheet. If stability data are unavailable, adopt conservative practices (store frozen, protected from light, use freshly thawed aliquots).
• Disposal: Collect DMSO-containing waste and dispose of according to institutional and local regulations.

For any lot-specific reconstitution solvent recommendations, concentration limits, or stability windows, consult the accompanying CoA and SDS.

Structure and Identity

Brief overview: Hesperadin is a bioactive small molecule supplied as part of Aladdin's Moligand™ collection for research and screening.

• Item-specific (from Product Data):
  • SKU: H409181
  • CAS: 422513-13-1
  • Grade: Moligand™
  • Storage: Store at −80°C
  • Shipped on: Dry ice packs + cold packs
• Not specified for this item; refer to CoA/Spec Sheet:
  • Molecular formula
  • Molecular weight
  • SMILES
  • InChIKey
• Literature/general notes (for identity context only; verify against primary references before use):
  • Hesperadin is widely cited in chemical biology literature as a small-molecule inhibitor used to interrogate mitotic kinase pathways. Structural descriptions in the literature depict a heteroaromatic, polycyclic scaffold incorporating multiple ring systems and hydrogen-bond acceptors/donors typical of ATP-competitive kinase ligands. Because multiple salt forms and vendor-specific identifiers may exist, confirm exact structure from the certificate of analysis prior to quantitative work.
• 2D structure description (general, literature-level):
  • A fused heteroaromatic core bearing donor/acceptor functionality and one or more anilide/arylamine linkages are commonly reported for compounds in this class, providing hinge-binding motifs and substituents that address the kinase ATP site. The exact atom connectivity, stereochemistry (if any), and counterions must be confirmed from the CoA for this specific lot.

Note: Do not use literature depictions as a substitute for the CoA/Spec Sheet when performing calculations or regulatory documentation.

Synthetic Utility

This material is provided as a finished small-molecule ligand for biological research. It is not commonly employed as a synthetic reagent or building block.

General synthetic chemistry perspective (informational):

• Structural class: Literature depictions of compounds named Hesperadin show heteroaromatic, kinase-directed scaffolds engineered for ATP-site interactions. Such scaffolds often arise from convergent syntheses joining heteroaryl cores and anilide/amine fragments via C–N bond formation (e.g., Buchwald–Hartwig amination), nucleophilic aromatic substitution (SNAr) on activated heteroaryl chlorides, and amide coupling steps.
• Retrosynthetic themes: Disconnection into (i) a hinge-binding heteroaryl fragment and (ii) a solubilizing/aromatic appendage is typical in medicinal chemistry routes, enabling SAR exploration through late-stage diversification.
• Derivatization potential: While the catalog item is not intended as a reagent, experienced chemists may draw inspiration from the scaffold to design analogs or tagged derivatives for target engagement studies (e.g., biotinylation, photoaffinity probes). Such modifications should be synthesized de novo; do not alter the supplied material unless analytically requalified.

If you require building blocks or catalysts for assembling related scaffolds, browse Aladdin’s heteroaryl halides, amide coupling reagents, and cross-coupling ligands.

Target Specificity

Item-specific target, selectivity, and potency details are Not specified for this item; refer to CoA/Spec Sheet and primary literature.

General note:

• Compounds known in the literature under the name Hesperadin have been used as tool molecules in kinase-focused studies. However, target profiles, isoform selectivity, and cellular potency can vary with assay conditions, cell type, and compound form (salt vs free base). Users should not assume equivalence across suppliers or lots.

User guidance:

• Determine the working concentration range empirically using dose–response analyses in your specific assay.
• Where feasible, complement biochemical IC50 data with cellular target engagement (e.g., phospho-biomarker modulation) and genetic rescue/knockdown controls to substantiate on-target effects.

No claims are made here regarding specific biological targets for this catalog item.

よくある質問

What is Moligand??

Moligand? is one of Aladdin Scientific's premium product lines, formulated for high-purity applications across multiple workflows. Compared to standard grades in the same workflow, Moligand? products typically offer tighter specifications, more rigorous QC, and stronger lot-to-lot consistency.

How should this product be stored?

Store at ?80 °C. Ultra-low temperature storage is required to maintain the specified shelf life.

How is this product shipped?

This product ships frozen with dry ice and cold packs. Unpack on arrival and transfer it to the storage condition stated above; handle dry ice with insulated gloves in a ventilated area.

What are the CAS number, molecular formula and molecular weight?

The CAS Number is 422513-13-1, the molecular formula is C29H32N4O3S, and the molecular weight is 516.67 g/mol.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.