DGKα-IN-2 - ≥98% , CAS No.2648556-92-5

CAS: 2648556-92-5 Cat. No.: D1440569 Formula: C₂₃H₂₂F₃N₃O₄ Molecular Weight: 461.43

GRADE & PURITY ≥98%

Storage

Store at 2-8°C

Shipped In

Wet ice

Overview

DGKα-IN-2 (example 48) is a DGKα inhibitor with the IC 50 of 0.9 nM, extracted from patent WO2021105115. DGKα-IN-2 significantly enhances the anti-tumor effect of anti-PD-1 by increasing the proliferation and function of T cells. DGKα-IN-2 has the potential for cancer and immunology study.

Specifications

Specifications & Purity

≥98%

Storage

Store at 2-8°C

Shipped In

Wet ice

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

Action Type

INHIBITOR

Purity

≥98%

Names and Identifiers

Molecular Weight

461.43

Application Protocols

No item-specific validated protocols are provided. The following is general guidance for small-molecule DGKα inhibitor evaluation and should be adapted to your system.

Example workflow (general, not item-specific):

1. Stock and plating:

• Dissolve compound at 10–50 mM in dry DMSO. Prepare serial dilutions in DMSO to achieve desired assay concentrations upon 100–1000× dilution into buffer/medium.

2. Biochemical DGKα activity assay:

• Recombinant DGKα with DAG substrate presented in mixed micelles or liposomes; include ATP/Mg2+.
• Incubate compound for 10–15 min pre-equilibration, then start reaction. Quench at defined time and quantify PA by LC–MS/MS or use a coupled ADP detection kit.

3. Cell-based signaling readout:

• Treat cells with compound or vehicle for 0.5–4 h. Measure pathway biomarkers (e.g., PKC substrate phosphorylation) and/or lipid species by LC–MS lipidomics. Include cytotoxicity control (e.g., CellTiter) and vehicle control.

4. Data analysis:

• Fit concentration–response curves; report apparent IC50 with 95% CI and Hill slope. Confirm reproducibility across biological replicates.

Notes:

• Control DMSO at constant final percent across wells.
• Check for precipitation or adsorption; use low-bind plastics.
• Validate with an orthogonal DGKα perturbation (genetic or alternative inhibitor) for mechanistic confidence.

Biological Roles

Context (general, not item-specific):

• Diacylglycerol kinases (DGKs) phosphorylate diacylglycerol (DAG) to phosphatidic acid (PA), controlling the balance of two bioactive lipids central to signaling and membrane biosynthesis. DGKα is a Ca2+-regulated isoform implicated in T-cell signaling, neuronal function, and cancer cell biology (literature).
• DAG acts as a second messenger activating protein kinase C (PKC) isoforms and RasGRP, while PA can modulate mTOR signaling, membrane curvature, and recruitment of signaling proteins.

Use of DGKα inhibitors in research (general guidance):

• Probe DGKα’s role in tuning DAG/PA pools and downstream signaling by transient chemical inhibition. Combine with lipidomics to confirm on-target effects.
• Investigate feedback control in receptor signaling (e.g., TCR, GPCRs) where localized DAG/PA dynamics shape signal amplitude and duration.
• Study membrane trafficking and cytoskeletal interfaces influenced by PA-binding proteins.

Assay readouts and validation:

• Biochemical assays: Monitor ATP consumption or PA formation using coupled enzyme or LC–MS readouts.
• Cell-based assays: Assess phosphorylation states of PKC substrates, ERK activation, or mTOR pathway markers as pathway-sensitive surrogates; confirm specificity with genetic controls.

Important note:

• No item-specific potency/selectivity data are provided for DGKα-IN-2 in this listing. Avoid biological over-interpretation without orthogonal validation. This product is supplied strictly for research use only.

Buffer Applications

DGKα-IN-2 is not a buffering reagent. However, buffers are integral to its use in biochemical and cell-based assays. The following guidance is general (not item-specific) and should be validated experimentally.

Typical assay buffers (general):

• Biochemical DGK assays: HEPES or Tris buffers (pH 7.2–7.5), Mg2+ for ATP-dependent catalysis, detergents or mixed micelles (e.g., Triton X-100 alternatives) or liposomes to present DAG substrates; include BSA if needed to stabilize hydrophobic components.
• Cell-based assays: Culture medium with defined serum; final DMSO ≤0.1–0.5% v/v. Use HEPES-buffered RPMI/DMEM for live-cell imaging.

Solubilization notes:

• Prepare concentrated DMSO stocks; dilute into buffer/medium with vigorous mixing to avoid precipitation. Evaluate adsorption to plastics; consider low-bind plates/tubes.

Controls and stability:

• Include vehicle controls (matching DMSO), and time-matched controls to account for potential compound instability in aqueous buffers.
• If preincubation with enzyme is required, keep consistent preincubation times/temperatures across conditions.

Do not use this product as a pH buffer. Select buffering systems based on the biological assay requirements and compatibility with lipid substrates and detection methods.

Green Alternatives

This item is a specialized research tool compound rather than a process solvent or commodity reagent, so green-chemistry substitution frameworks are not typically applicable.

General considerations for greener practice when handling bioactive small molecules (not item-specific):

• Minimize DMSO consumption by preparing concentrated stocks and performing serial dilutions to working concentrations immediately prior to use.
• Employ microscale assays (384/1536-well formats) to reduce compound, solvent, and plastic use.
• Choose aqueous buffers over organic-rich media wherever assay performance allows, and validate low DMSO (≤0.1–0.5% v/v) conditions.
• Optimize storage to reduce waste from degradation (aliquot to avoid repeated freeze–thaw and light exposure; see Storage tab).
• For any synthesis or analog work, consider greener solvents (e.g., 2-MeTHF, CPME, EtOAc) and catalytic methods, guided by solvent selection guides from ACS PMI/Pharma.

Trade-offs:

• Solubility and stability of lipophilic inhibitors often necessitate DMSO; greener substitutes may compromise assay reliability. Validate performance against your critical endpoints before implementing changes.

Pharmaceutical Uses

No pharmaceutical or clinical uses are claimed or supported for this product. It is provided strictly for research use only.

Context (general, not item-specific):

• Small-molecule enzyme inhibitors like DGKα tool compounds may be explored in discovery research to understand target biology. Such materials are not drug products, APIs, or excipients and are not suitable for human or veterinary use.
• There is no pharmacopeial monograph or excipient designation provided for this item.

Laboratory and discovery relevance:

• Useful as a chemical probe candidate in target validation workflows, contingent on users establishing potency, selectivity, and exposure in their specific systems.
• Not for diagnostic procedures, clinical applications, or manufacturing of medicinal products.

Physical Properties

Item-specific physicochemical properties were not supplied for this listing.

Item-specific values (from 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.
• Melting point (MP): Not specified for this item; refer to CoA/Spec Sheet.
• Boiling point (BP): Not applicable/rarely reported for non-volatile bioactive solids; Not specified for this item; refer to CoA/Spec Sheet.
• Density: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Not specified for this item; refer to CoA/Spec Sheet.
• LogP/logD, pKa, refractive index: Not specified for this item; refer to CoA/Spec Sheet.

General characterization guidance (literature-level, not item-specific):

• Many kinase-directed small molecules are crystalline or amorphous solids with limited aqueous solubility and good solubility in polar aprotic solvents (DMSO, DMF) and certain organic solvents (MeCN, EtOAc, alcohols).
• If required, determine aqueous solubility empirically across assay-relevant buffers (pH 6.5–7.5) and with co-solvent screening (e.g., 0.1–1% DMSO in buffer). Consider kinetic vs thermodynamic solubility.
• pKa and logD (pH 7.4) influence cell permeability and target engagement; measure via potentiometric titration or shake-flask/LC methods.
• Thermal behavior can be assessed via DSC/TGA to inform handling and storage.

Analytical quality checks (general guidance):

• LC–MS purity and retention time, UV spectrum, and NMR confirmation of structure are recommended prior to biological assays.
• If polymorphism is suspected, PXRD can help ensure batch-to-batch consistency for screening work.

Quality & Grades

Item-specific grade/purity were not provided for this listing. Refer to the product’s CoA/Spec Sheet for the precise assay purity and analytical methods used.

Item-specific details (from Product Data):

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

General notes on grades (for small-molecule screening tools):

• Screening/Research grade: Typically provided with LC–MS or HPLC purity and identity confirmation (LC–MS, 1H NMR, sometimes HRMS). Suitable for target validation, biochemical assays, and cell-based studies when supported by in-house QC.
• HPLC/Chromatography grade (solvents): Not applicable here.
• Stabilizers: None listed. If a stabilizer or counterion is used, it will be indicated on the CoA. Absence of a stabilizer does not imply stability; perform periodic QC for long programs.

What to check on receipt (practical guidance):

• Confirm purity by LC–UV/MS in your own system under assay-relevant conditions; check for late-eluting impurities.
• Verify identity by MS and NMR against literature where available.
• Record lot/batch, appearance, and any hygroscopic or adhesive behavior. If needed, dry under vacuum at ambient temperature and promptly re-seal in a desiccator.

Suitability notes:

• For quantitative biology, ensure consistent solvent system (e.g., DMSO lot) and account for any salt form or residual solvents reported on the CoA.

Reaction & Applications

This product is a small-molecule research tool (DGKα inhibitor candidate) rather than a synthetic reagent. Consequently, classic synthetic reaction applications (e.g., cross-coupling, nucleophilic substitutions) are not the primary use case.

Research applications (general, not item-specific):

• Target engagement studies: Evaluate modulation of diacylglycerol (DAG) to phosphatidic acid (PA) conversion catalyzed by DGKα in biochemical assays.
• Cellular pathway interrogation: Probe DAG/PA-regulated signaling pathways (e.g., PKC, mTOR, RasGRP) by pharmacological inhibition of DGKα in cell models.
• Lipidomics readouts: Quantify DAG/PA species via LC–MS/MS to confirm on-target lipid changes.
• Chemical biology controls: Pair with orthogonal DGKα inhibitors or genetic perturbations (siRNA/CRISPR) for mechanistic validation.

If using chemically (synthetic context):

• Not typically employed as a building block or catalyst. Any derivatization should be preceded by full structure confirmation and IP review.

Important note: No item-specific potency, selectivity, or kinetic constants are provided here. Users should establish concentration–response relationships, time-dependence, and reversibility in their system of interest and consult primary literature for DGKα assay designs.

Reaction Conditions

Not typically applicable. DGKα-IN-2 is supplied as a small-molecule bioactive tool and is not intended for use as a reagent or catalyst in synthetic transformations.

General experimental conditions for biological evaluation (not item-specific):

• Stock preparation: Commonly 10–50 mM in anhydrous DMSO; verify solubility and stability.
• Working concentrations: Establish a concentration–response curve bracketing expected activity; confirm absence of aggregation (e.g., detergent sensitivity tests, dynamic light scattering) and non-specific interference.
• Temperature/time: Many biochemical assays run at 25–37°C for 10–60 minutes; cell-based treatments range from minutes to 24–72 hours depending on endpoints.
• Controls: Include vehicle and orthogonal tool controls; consider counterscreens against related DGK isoforms or off-target kinases as available.

These are general guidelines and are not item-specific conditions. Refer to primary literature for DGKα assay designs and optimize per your biological system.

Safety & Handling

Item-specific hazard classification was not provided for this product. Treat as a research chemical of unknown toxicity and consult the Safety Data Sheet (SDS) upon receipt.

Item-specific safety data (from Product Data):

• GHS classification: Not specified for this item; refer to SDS.
• Signal word: Not specified for this item; refer to SDS.
• Hazard statements (H-codes): Not specified for this item; refer to SDS.
• Pictograms: Not specified for this item; refer to SDS.

General laboratory precautions (good practice for small-molecule inhibitors):

• Handle in a fume hood; avoid inhalation, ingestion, and skin/eye contact. Use standard PPE: lab coat, safety glasses, and nitrile gloves; upgrade to splash goggles and double-gloving during weighing or solution prep.
• Avoid dust/aerosol generation. Use antistatic spatulas and weigh boats; cap vials promptly.
• Incompatibilities: Strong oxidizers and strong acids/bases may degrade typical organic small molecules. Keep away from ignition sources and high heat.
• First aid (general): In case of skin contact, wash with soap and water; for eye exposure, rinse with water for 15 minutes and seek medical attention; if inhaled, move to fresh air; if ingested, seek medical attention. Always follow SDS for definitive guidance.
• Spill response: Contain with inert absorbent, collect in compatible waste. Decontaminate surfaces with appropriate solvent/detergent.

Waste disposal:

• Collect in organic hazardous waste per institutional and regulatory requirements. Do not discharge to drains.

Solvent Selection

Item-specific solubility data are not available for this product; consult the CoA/Spec Sheet. The guidance below reflects common practice for small-molecule kinase/in-lipid enzyme inhibitors and should be verified experimentally for DGKα-IN-2.

General solvent strategy (literature-level, not item-specific):

• Stock solutions: Many screening compounds are dissolved in anhydrous DMSO at 10–50 mM, then diluted into aqueous assay buffers to a final 0.1–1% DMSO (v/v). Confirm absence of precipitation after dilution.
• Alternative solvents: If DMSO is unsuitable, consider DMF, 1-methyl-2-pyrrolidone (NMP), or mixtures with co-solvents (MeCN, EtOH, PEG400). Evaluate cytotoxicity of co-solvents in cell assays.
• Buffer compatibility: Use HEPES or PBS-based buffers; include low levels of non-ionic surfactant (e.g., 0.01–0.05% Tween-20) only if needed to maintain solubility, after confirming no assay interference.

Polarity and miscibility (general):

• DMSO is polar aprotic and fully miscible with water; it generally maintains compound stability and minimizes hydrolysis relative to aqueous storage.
• Avoid strong acids/bases unless stability is confirmed.

Practical tips:

• Warm gently (25–37°C) and sonicate to aid dissolution; avoid prolonged heating.
• Filter 0.22 µm if particulate is observed (ensure no adsorption losses).
• Document precipitation limits by visual inspection and by LC–UV after dilution into the intended assay matrix.

Storage & Reconstitution

Item-specific handling (from Product Data):

• Storage conditions: Store at 2–8°C.
• Shipped in: Wet ice.

Additional guidance (general best practice, not item-specific):

• Protect from moisture and light; keep container tightly closed. If the compound is hygroscopic or photosensitive (not specified), handle quickly under low humidity and minimize light exposure.
• Upon first opening, allow vial to equilibrate to room temperature in a desiccator before unsealing to reduce moisture condensation.

Reconstitution (general):

• Prepare a concentrated stock in anhydrous DMSO (e.g., 10–50 mM) immediately before use. Mix by gentle vortex and brief sonication if needed. Filter through 0.22 µm only if particulates persist and adsorption is minimal.
• Aliquot stock solutions in small volumes to avoid repeated freeze–thaw. Many labs store DMSO stocks at −20°C or −80°C for medium-term use; confirm stability empirically. Note: This is general guidance and does not override the item’s labeled storage at 2–8°C for the solid form.

Stability monitoring:

• Periodically check by LC–MS/HPLC for degradation or precipitation. Discard if significant impurity growth or activity loss is observed.

Always refer to the CoA/Spec Sheet and SDS for definitive, lot-specific handling and stability information.

Structure & Identity

Overview: DGKα-IN-2 is described as a small-molecule inhibitor tool compound targeting diacylglycerol kinase alpha (DGKα). Item-specific structural identifiers are not provided in the current listing.

Item-specific identifiers (from Product Data):

• CAS: 2648556-92-5
• 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.

General structure notes (literature-level, not item-specific):

• DGKα inhibitors often incorporate heteroaromatic or bicyclic scaffolds that enable ATP-site or lipid-substrate-site engagement within DGKα, with H-bond donor/acceptor arrays and lipophilic features to recognize the catalytic domain.
• Typical kinase/in-lipid enzyme inhibitors include amide, urea, or biaryl linkages to balance polarity and membrane permeability; however, the exact functional groups for DGKα-IN-2 are not specified here.

2D structure description: Not provided for this specific item. In the absence of SMILES/InChI, users should consult the Certificate of Analysis (CoA) or Spec Sheet to confirm structural features and identifiers prior to experimental planning or analytical qualification.

Practical identity confirmation (general guidance):

• Verify by LC–MS (single, high-abundance [M+H]+), 1H/13C NMR, and HRMS when first received.
• Cross-reference CAS and any available literature/source entries for DGKα-IN-2 to ensure alignment with analytical fingerprints before biological testing.

Synthetic Utility

DGKα-IN-2 is intended as a bioactive research tool and is not typically employed as a synthetic reagent, monomer, or building block.

Non-applicability notes:

• No functional-group handle or reactivity profile is provided, and no item-specific synthetic transformations are recommended.

General advice (if derivatization is contemplated for SAR or probe development):

• Confirm full structure and purity before any modification. Map potential diversification vectors via NMR/LC–MS and, where available, literature precedent.
• Consider late-stage diversification strategies (amide coupling, Suzuki/Negishi for biaryl elaboration, urea/carbamate formation) only if compatible with the actual scaffold once known from the CoA/structure.
• Track effects of modifications on solubility, permeability, and metabolic stability; apply microscale parallel synthesis with green solvent choices where feasible.

For routine chemistry workflows, prioritize reagents specifically designed for building-block utility; this product is best deployed in biological assays rather than in preparative synthesis.

Target Specificity

Item-specific target specificity data (e.g., Ki, IC50, isoform selectivity, off-target panels) are not provided for this listing.

Non-applicability of antibody-style fields:

• Antigen/epitope/clone/isotype fields are not relevant to this small-molecule product.

General guidance for establishing specificity (not item-specific):

• Biochemical profiling: Determine potency against DGKα using purified enzyme and measure activity versus related DGK isoforms (β, γ, δ, etc.) to assess selectivity.
• Cellular target engagement: Apply CETSA, DARTS, or NanoBRET target engagement assays. Complement with lipidomics to confirm DAG→PA modulation.
• Off-target assessment: Screen against representative kinases/lipid kinases or use broad kinase panels when feasible to identify liabilities.
• Genetic orthogonality: Compare pharmacological effects with DGKα knockdown/knockout to support on-target action.

Until item-specific data are obtained, treat DGKα-IN-2 as an uncharacterized probe candidate and interpret biological results cautiously.

Frequently Asked Questions

What is the purity of this product?

This product is supplied at ≥98% purity (chemical assay). Lot-specific values are stated on the Certificate of Analysis.

How should this product be stored?

Store at 2–8 °C. Refrigerated storage is required to maintain the specified shelf life.

How is this product shipped?

This product ships chilled on wet ice. Unpack on arrival and transfer it to the storage condition stated above.

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

The CAS Number is 2648556-92-5, the molecular formula is C23H22F3N3O4, and the molecular weight is 461.43 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.