A-674563 hydrochloride - ≥99% , CAS No.2070009-66-2

CAS: 2070009-66-2 Cat. No.: A649264 Summenformel: C₂₂H₂₃ClN₄O Molekulargewicht: 394.90

GRADE & PURITY ≥99%

Storage

Store at 2-8°C

Shipped In

Wet ice

Übersicht

A-674563 hydrochloride is a potent and selective Akt1 inhibitor with K i of 11 nM.

In Vitro

A-674563 slows proliferation of tumor cells with an EC 50 of 0.4 μM. A563 (0-10 µM) significantly decreases GSK3 and MDM2 phosphorylation in STS cells. A563 shows inhibitory effect on all STS cell lines, with IC 50 values at 48 hours ranging from 0.22±0.034 µM (SW684) to 0.35 ±0.06 µM (SKLMS1). A563 induces G2 cell cycle arrest and apoptosis in STS cells. A563 (1 µM/12 hr) upregulates the expression of GADD45A independent of p53. A-674563 (10-1000 nM) is anti-proliferative and cytotoxic in cultured human melanoma cells, induces melanoma cell apoptotic death, inhibited by caspase inhibitors, and inhibits melanoma cells via Akt-dependent and -independent mechanisms. A-674563 is cytotoxic and anti-proliferative when added to U937 and AmL progenitor cells, activates caspase-3/9 and apoptosis in U937 and AmL progenitor cells, and manipulates other signalings in AmL cells whiling blocking Akt . MCE has not independently confirmed the accuracy of these methods. They are for reference only.

In Vivo

A-674563 (40 mg/kg/d, p.o.) shows no significant monotherapy activity, but the efficacy of the combination therapy (A-674563+paclitaxel) is significantly improved in the PC-3 prostate cancer xenograft model. A-674563 (20, 100 mg/kg) increases plasma insulin in an oral glucose tolerance test . A563 (20 mg/kg/bid; p.o.) exhibits slow tumor growth and a significant difference in tumor volume without significant weight loss of mice. A563-treated tumors express increased levels of GADD45α and decreased levels of PCNA (a nuclear marker for proliferation). Additionally, TUNEL assay staining levels (marker for apoptosis) increase in the A563-treated specimens. A-674563 (25, 100 mg/kg, lavage daily) potently inhibits A375 xenograft growth in mice. A-674563 (15, 40 mg/kg) injection inhibits U937 xenograft in vivo growth, and improves mice survival. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Form:Solid

IC50& Target:Akt1 11 nM (Ki) PKA 16 nM (Ki) CDK2 46 nM (Ki) GSK3β 110 nM (Ki) ERK2 260 nM (Ki) PKCδ 360 nM (Ki) RSK2 580 nM (Ki) MAPK-AP2 1.1 μM (Ki) PKCγ 1.2 μM (Ki) Chk1 2.6 μM (Ki) CK2 5.4 μM (Ki) SRC 13 μM (Ki)

Specifications

Spezifikationen & Reinheit

≥99%

Biochemische und physiologische Mechanismen

A-674563 hydrochloride is a potent and selective Akt1 inhibitor with K i of 11 nM.

Storage

Store at 2-8°C

Verschickt in

Wet ice

Dieses Produkt erfordert Kühlkettenversand. Grundversand und andere Economy-Optionen sind nicht verfügbar.

Aktionsart

INHIBITOR

Reinheit

≥99%

Namen und Kennungen

• Molekulargewicht: 394.90

Chemische und physikalische Eigenschaften

• Löslichkeit: H2O : 100 mg/mL (253.23 mM; Need ultrasonic) DMSO : 62.5 mg/mL (158.27 mM; Need ultrasonic)

Application Protocols

No tested application protocols are provided in the Product Data for this item. General, non-binding guidance for small-molecule screening compounds:

• Stock preparation: Dissolve in anhydrous DMSO to 10–50 mM. Vortex and, if necessary, briefly sonicate. Filter if particulate persists.
• Working solutions: Dilute into assay buffer to the desired final concentration, keeping DMSO constant across all wells/samples (typically 0.1–1% v/v).
• Controls: Include vehicle controls and, where applicable, a known positive/negative control appropriate to the assay.
• Plate handling: Use low-binding plates or include a small amount of nonionic detergent (e.g., 0.01% Tween 20) to minimize adsorption.
• Storage: Aliquot stocks to single-use volumes; avoid repeated freeze–thaw cycles. These steps are offered as general best practices and are not validated for this specific item. For any validated, item-specific protocols, refer to the CoA/Spec Sheet or contact Technical Support.

Biological Roles

• Category context: This product is listed under small molecules/compound libraries for research use. Compounds in this class are commonly used as chemical probes or reference standards for interrogating cellular signaling and enzymatic pathways (general literature context).
• Functional role (general): Many hydrochloride salts of bioactive amines act as modulators of enzyme activity, receptors, ion channels, or signaling kinases in cell-free and cellular assays. The salt form often improves handling and aqueous compatibility without altering the intrinsic mode of action of the free base.
• Recommended research practices:
  • Establish a clear concentration–response relationship (8–12 point curves) in the relevant assay system.
  • Control for cytotoxicity or off-target effects using orthogonal readouts and appropriate vehicle controls.
  • Verify on-target pathway modulation using secondary assays (e.g., phosphorylation status, reporter genes, or biophysical binding), recognizing that such validation is compound- and target-specific.
• Data integrity:
  • Record stock concentration, solvent, and lot number.
  • Confirm stability and absence of aggregation or fluorescence interference where optical readouts are used (e.g., include detergent or use counter-screens).
• Important: No medical, diagnostic, or in vivo claims are made. This product is intended strictly for laboratory research.

Buffer Applications

This item is a research small molecule rather than a buffering agent. Therefore, classical buffer system formulation (pKa-based buffering capacity) is not directly applicable. Practical notes for assay buffers (general guidance):

• Dissolution: If soluble, dissolve directly into mildly acidic aqueous buffers (e.g., acetate or citrate, pH 4–6) to maintain the protonated amine form; otherwise introduce from a DMSO stock into the working buffer with vigorous mixing.
• Common assay buffers: HEPES, Tris, PBS, or MOPS at standard concentrations (25–50 mM), supplemented with 0.01–0.05% nonionic detergent to mitigate nonspecific adsorption if needed.
• Compatibility: Validate that the compound does not precipitate upon dilution and that buffer components (e.g., high phosphate) do not form insoluble ion pairs.
• Filtration: If sterile conditions are required, filter the final working solution (0.22 µm) after dilution; avoid filtering highly concentrated organic stocks.

Green Alternatives

For this class of small-molecule hydrochloride salts, environmental impact is driven less by the compound itself and more by solvent choice and workflow design. Consider the following greener practices:

• Prefer aqueous media where feasible:
  • If the hydrochloride form is sufficiently water-soluble, formulate directly in buffered saline at mildly acidic pH, avoiding high DMSO fractions.
• Minimize DMSO load:
  • Use concentrated DMSO stocks and dilute to ≤0.5% v/v in assays when compatible; this reduces VOC burden and waste.
• Solvent alternatives (comparison; general guidance):
  • Water/buffer (pH 4–6): Greenest option when soluble; check stability.
  • Ethanol (Ph. Eur./USP grade): Biobased alternative for analytical dilutions; typically limit to low percentages in bioassays.
  • 2-propanol: More benign than chlorinated solvents for rinsing and cleanup.
• Operational improvements:
  • Microscale assays and miniaturized plates (384/1536-well) reduce solvent and compound consumption.
  • Closed, nitrogen-backed storage of DMSO stocks extends shelf-life and reduces degradation/waste.
• Waste management:
  • Segregate halogen-free organic waste from aqueous streams; avoid chlorinated solvents unless strictly required. Note: No item-specific green certifications are provided. Adopt institution-specific green chemistry guidelines for solvent selection and waste handling.

Pharmaceutical Uses

• Regulatory/compendial status: Not specified for this item; refer to CoA/Spec Sheet. This product is sold for research use only and is not manufactured under GMP for human use.
• Typical roles in pharma R&D (general):
  • Reference standard for analytical method development (HPLC/LC–MS quantification, stability-indicating methods).
  • Tool compound in target validation and SAR benchmarking during lead discovery.
  • Positive control in biochemical/cellular assays to establish assay window and system suitability.
• Formulation research (in vitro only):
  • Prepare concentrated DMSO stocks for screening; for biophysical assays, explore aqueous buffers, cyclodextrin inclusion, or mild acidification to enhance solubility as appropriate for hydrochloride salts.
• Impurity/degradation tracking: Establish forced-degradation profiles (acid/base, oxidative, thermal, photolytic) to understand analytical behavior; this supports robust quantification in preclinical research. Note that any degradation studies are for laboratory knowledge-building only, not for clinical development, unless conducted under appropriate quality systems.
• Important: No therapeutic, diagnostic, or clinical application is implied. Not for human or veterinary administration.

Physical Properties

Item-specific physicochemical data have not been provided for this catalog entry. Where applicable, use the following guidance:

• 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 applicable for an ionic salt (decomposes) unless otherwise reported; item-specific value not provided.
• Density: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Not specified for this item; refer to CoA/Spec Sheet. General note (literature): hydrochloride salts of amine-containing drug-like molecules are commonly soluble in polar media (water, aqueous buffers, methanol, DMSO) to varying degrees; establish actual solubility empirically for this item.
• logP/logD, pKa, refractive index: Not specified for this item; refer to CoA/Spec Sheet.
• Hygroscopicity: Many amine hydrochlorides are mildly hygroscopic; verify handling needs experimentally for this item. Practical tips:
• Prepare an initial 10 mM stock in anhydrous DMSO, then serially dilute into assay buffer; confirm absence of precipitation visually and by UV/LC.
• If aqueous solubility is limited, co-solvents (≤1–2% DMSO or ethanol in buffer) or pH adjustment (acidic media to keep amine protonated) may help (general guidance).

Quality and Grades

• Grade/purity: Not specified for this item; refer to CoA/Spec Sheet.
• What to expect on CoA: Identity confirmation (typically LC–MS and/or NMR), purity by HPLC/UPLC (% area), counter-ion content (chloride), water content (Karl Fischer), and residual solvents as applicable. Only trust the lot-specific CoA as the binding specification document.
• Implications of hydrochloride form: The HCl salt generally provides improved batch-to-batch handling, increased aqueous solubility, and defined stoichiometry compared to the free base. Verify equivalence (free-base conversion) when preparing concentration-normalized stocks.
• Typical analytical acceptance for screening compounds (general guidance; not item-specific specs):
  • HPLC purity ≥95% (area %)
  • Identity consistent with reference spectrum
  • Residual solvents within ICH-reasonable limits
  • Water content appropriate for a crystalline salt
• If UV transparency is critical (e.g., biochemical assays), request UV cut-off and impurity profile on the CoA; do not assume HPLC-grade optical clarity unless explicitly stated.
• Stabilizers: Not specified for this item; refer to CoA/Spec Sheet. If none are used, store as supplied and minimize exposure to heat, light, and humidity.

Reaction and Applications

This product is supplied as a research-grade small molecule for screening and biochemical/biological research, not primarily as a synthetic reagent. Accordingly, classical synthetic reaction applications (e.g., cross-couplings, Grignards) are not typically relevant to this item.

• Typical research applications (general for bioactive tool compounds):
  • Use as a reference or tool compound in cell-free enzymatic assays, cell-based signaling readouts, or phenotypic screens.
  • Benchmarking assay windows, Z′-factor validation, and plate-to-plate controls.
  • Mechanistic studies in pathway interrogation, target engagement (e.g., CETSA, DARTS), and structure–activity relationship comparisons with analogs.
• Practical guidance:
  • Prepare concentrated DMSO stocks (e.g., 10 mM), aliquot to avoid freeze–thaw, and verify concentration by UV or quantitative NMR when accuracy is critical.
  • Assess compound stability in assay media over the assay timeframe (LC–MS check for degradation or adsorption to plastics).
  • Include vehicle controls matching the final DMSO content across all wells/samples.
• Note: If you intend to derivatize or immobilize the molecule for chemoproteomics, consult structural information from primary literature or the CoA; site-selective handles (amines/phenols) and their accessibility are compound-specific and not provided here.

Reaction Conditions

Classical synthetic reaction conditions are not generally applicable to this product as sold (a bioactive small-molecule hydrochloride for screening). If derivatization is planned, conditions must be tailored to the disclosed structure from primary references or the CoA. General, non-item-specific guidance for handling small-molecule HCl salts in chemistry workflows:

• Free-basing: Dissolve the salt in minimal water or methanol, add saturated NaHCO3, and extract with an organic solvent (EtOAc/MTBE) to obtain the free base (verify by LC–MS). Alternatively, ion-exchange resins can be used.
• Couplings: Conduct amide/carbamate formation in anhydrous polar aprotic solvents (DMF, DCM, MeCN) with standard peptide coupling reagents (HATU, EDCI) if a primary/secondary amine is present (structure-dependent).
• Stability: Avoid strong base or prolonged high temperature if the scaffold contains base-labile motifs; avoid strong oxidants that can chlorinate or oxidize heteroaromatics.
• Purification: Reverse-phase preparative HPLC is often preferred for drug-like molecules; form the desired salt (e.g., HCl) during isolation for consistent handling. These conditions are provided as generic literature guidance for amine-containing small molecules and are not specific to this catalog item.

Safety and Handling

• GHS classification: Not specified for this item; refer to SDS.
• Signal word / H-statements / pictograms: Not specified for this item; refer to SDS for authoritative hazard communication.
• Likely hazards (general for bioactive small molecules): May cause irritation to skin, eyes, and respiratory tract; avoid inhalation of dust and contact with skin/eyes. Handle in a fume hood.
• PPE: Lab coat, safety glasses, and appropriate chemically resistant gloves (e.g., nitrile). Use dust control (weighing cone) and micro-spatulas.
• Storage incompatibilities: Keep away from strong oxidizers and strong bases that can neutralize the hydrochloride. Avoid moisture if material is hygroscopic.
• First aid (overview; consult SDS):
  • Inhalation: Move to fresh air; seek medical attention if symptoms persist.
  • Skin: Wash with soap and water; remove contaminated clothing.
  • Eyes: Rinse cautiously with water for several minutes; remove contact lenses if present and easy; seek medical advice if irritation continues.
  • Ingestion: Rinse mouth; seek medical attention.
• Spill response: Avoid dust; collect mechanically; wipe residues with damp disposable towels and appropriate detergent; dispose according to local regulations.
• SDS: Always consult the product-specific Safety Data Sheet for definitive hazard, toxicological, and disposal information.
• Research use only: Not for human or veterinary use.

Solvent Selection

Because the exact solubility parameters for this item are not provided, select solvents empirically with a tiered approach suitable for amine hydrochloride salts:

• Primary stock solvent (screening workflows):
  • DMSO (anhydrous): Generally affords high solubility for diverse small molecules; prepare 10–50 mM stocks, store aliquots at ≤–20°C to minimize freeze–thaw.
  • Water or aqueous buffers (pH 2–6): Protonated amines remain in solution; verify clarity and stability.
• Secondary/co-solvents for formulation or assay introduction:
  • Methanol or ethanol (analytical sample prep), followed by dilution into buffer (final organic ≤1–2%).
  • PEG400 or 10–20% cyclodextrin solutions for challenging solubility cases (biophysical assays).
• Miscibility and polarity (general): DMSO and lower alcohols are fully miscible with water and compatible with most biochemical assays at low percentages.
• Practical decision tree:
  1. Attempt dissolution at room temperature in DMSO.
  2. If aqueous delivery is required, dilute DMSO stock into buffer while vortexing; limit final DMSO to ≤0.5–1% unless assay tolerates more.
  3. If precipitation occurs, reduce concentration, adjust pH slightly acidic, or introduce a small co-solvent fraction.
• Comparators:
  • DMSO vs DMF: DMSO is preferred for bioassays due to lower volatility and toxicity; DMF can be used for analytical sample prep but is less favored in biology.

Storage and Reconstitution

• Storage conditions (as provided): Store at 2–8°C. Shipments are on wet ice.
• Long-term storage: For solid material, keep tightly sealed in the original container, protected from moisture and light. If preparing solutions, store aliquots at ≤–20°C unless solubility/stability data indicate otherwise (general guidance); avoid frost-free cycles.
• Reconstitution (general guidance for small-molecule HCl salts):
  • Bring container to room temperature in a desiccator before opening to avoid moisture condensation.
  • Prepare a concentrated stock solution in anhydrous DMSO or in appropriate aqueous buffer if directly soluble. Record the exact solvent and concentration.
  • Filter through 0.22 µm PTFE (organic) or PES (aqueous) syringe filters if particulate is observed.
• Stability after reconstitution: Not specified for this item; refer to CoA/Spec Sheet. In the absence of item-specific data, use within 1–2 weeks at 2–8°C or within 3–6 months at –20°C, minimizing freeze–thaw (general best practice).
• Compatibility: Avoid strong bases and oxidizers. Maintain mildly acidic to neutral pH in aqueous solutions to preserve the protonated form.
• Research use only: Not for human or veterinary use.

Structure and Identity

• Product name: A-674563 hydrochloride (small-molecule HCl salt; screening compound)
• CAS: 2070009-66-2
• 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 context): As an HCl salt, the compound contains at least one basic nitrogen (e.g., amine/aza-heterocycle) that is protonated to form the hydrochloride. The free base of such tool molecules often contains aromatic/heteroaromatic rings and H-bonding motifs to engage protein active sites (general literature context; not item-specific).
• 2D structure description (general): Typical hydrochloride salts of bioactive small molecules consist of an organic cation (protonated amine/heterocycle) paired with chloride anion; ring systems may be fused or biaryl, with polar heteroatoms enabling aqueous solubility upon protonation (general literature context).
• Identity confirmation: For definitive identity and lot-specific attributes (NMR, LC–MS, HPLC purity, salt content, counter-ion ratio), consult the item’s Certificate of Analysis (CoA).

Synthetic Utility

This product is not primarily intended as a synthetic intermediate or building block. Without a disclosed structure in this listing, specific retrosynthetic or functional-group-based transformations cannot be recommended. General considerations if derivatization is contemplated (literature-guided, not item-specific):

• Hydrochloride salts typically indicate the presence of a basic nitrogen; free-basing (e.g., with NaHCO3 or Na2CO3 in biphasic systems) can liberate the neutral amine for coupling chemistry.
• Potential late-stage functionalization strategies for drug-like scaffolds (if chemically compatible):
  • Amide formation (HATU/EDC) from primary/secondary amines.
  • N-alkylation or reductive amination to explore SAR.
  • Succinimidyl carbamate installation for immobilization/affinity capture.
• Analytical control: Monitor reactions by LC–MS; salts may require ion-exchange workups to recover the free base or re-form the hydrochloride. Because this entry lacks structural specifics, confirm exact reactive handles and protection/deprotection needs from the CoA or primary literature before attempting any synthetic modification.

Target Specificity

No target specificity data (e.g., biochemical target, pathway, or selectivity profile) are provided in the Product Data for this item. In keeping with Aladdin Scientific’s policy:

• Only item-specific target or assay validation information from our catalog/CoA would be listed here.
• For literature reports regarding potential biological targets of compounds with similar names, consult peer-reviewed sources independently; such information is not asserted or guaranteed for this specific lot/product. If you require confirmed target data, please request the latest CoA or technical dossier for this SKU (A649264).