Doxorubicin-SMCC - Moligand™, 10 mM in DMSO , CAS No.400647-59-8

CAS: 400647-59-8 Cat. No.: D1494975 Formula: C₃₉H₄₂N₂O₁₄ Peso molecolare: 762.8

GRADE & PURITY Moligand™, 10 mM in DMSO

Storage

Store at -80°C

Shipped In

Dry ice packs + Cold packs

Panoramica

Doxorubicin-SMCC is a agent-linker conjugate for ADC . Doxorubicin-SMCC contains a non-cleavable ADC linker and a DNA topoisomerase II inhibitor Doxorubicin.

Specifications

Specifiche e purezza

Moligand™, 10 mM in DMSO

Condizioni di conservazione di stoccaggio

Store at -80°C

Spedito in

Dry ice packs + Cold packs

Questo prodotto richiede spedizione a catena fredda. I servizi di terra e altri servizi economici non sono disponibili.

Grado

Moligand™

Nomi e identificatori

Peso molecolare

762.8

Application Protocols

Example workflow for thiol-directed conjugation of a cysteine-containing peptide/protein using Doxorubicin-SMCC (literature/general; optimize empirically):

1. Prepare stock: Dissolve Doxorubicin-SMCC in anhydrous DMSO at 10–20 mM. Store on ice protected from light until use.
2. Reduce target thiols (if needed): Treat the peptide/protein with 0.5–2 mM TCEP in 50 mM phosphate, 100 mM NaCl, 1 mM EDTA, pH 7.0 for 30 min at room temp. Desalt into conjugation buffer (pH 6.8) to remove reductant.
3. Conjugation: Mix target (e.g., 50 µM thiol) with Doxorubicin-SMCC at 1.2–2.0 equivalents per thiol. Final DMSO ≤10%. Incubate 30–90 min at 20–25°C, protected from light.
4. Quench: Add 5–10 mM cysteine or 2-mercaptoethanol, 10 min.
5. Purify: Desalt or perform SEC/HPLC to remove excess small molecules and confirm product by LC–MS or SDS-PAGE (doxorubicin fluorescence aids visualization).
6. Characterize: Determine substitution level by UV–Vis (using doxorubicin extinction coefficient, literature) or mass spectrometry; assess stability in relevant buffers.

Notes:

• Perform small-scale pilot reactions to determine optimal equivalents and time.
• Maintain gentle mixing and avoid foaming. Use amberware and minimize light exposure throughout.

Biological Roles

This section summarizes general, literature-based biochemical roles of the components; it does not imply any therapeutic use.

• Doxorubicin (literature): an anthracycline that intercalates into DNA and inhibits topoisomerase II, leading to disruption of DNA replication and transcription in proliferating cells. It can generate reactive oxygen species through redox cycling, and associates strongly with nucleic acids due to its planar chromophore and cationic aminosugar.
• SMCC linker (literature): a bifunctional succinimidyl ester–maleimide reagent widely used to bridge amines and thiols. In Doxorubicin-SMCC, the amine-reactive function is consumed to install the spacer on doxorubicin, typically leaving the maleimide free for thiol conjugation.
• Conjugate behavior (literature/general): the hydrophobic cyclohexyl spacer can influence membrane interactions and aggregation of bioconjugates; the doxorubicin chromophore enables optical tracking in biochemical assays (visible fluorescence and absorbance).

Research implications:

• Useful as a chemical biology probe to study carrier–payload delivery mechanisms, intracellular trafficking, and linker stability in vitro.

Note: All biological roles described are mechanistic and literature-based; this product is intended strictly for research use in laboratory settings.

Buffer Applications

Buffers are chosen to optimize maleimide–thiol coupling while preserving target integrity (literature/general). Item-specific buffer recommendations are not provided; follow your application needs and CoA guidance.

• Preferred pH: 6.5–7.0 to maximize cysteine selectivity and minimize maleimide hydrolysis.
• Avoid: primary amine buffers (e.g., Tris, glycine) that can add to maleimides; high pH (>7.5) which accelerates ring opening; reducing thiols (DTT, βME) during conjugation.
• Compatible buffers: phosphate (10–100 mM), HEPES, citrate-phosphate. Include 1–5 mM EDTA to chelate trace metals that can promote oxidation.
• Ionic strength: 50–200 mM NaCl helps maintain protein solubility and reduces nonspecific interactions.
• Organic cosolvent: ≤10% DMSO or DMF to dissolve hydrophobic linker–payloads; verify target tolerance.

Recipe sketch (example, literature):

• 50 mM sodium phosphate, 100 mM NaCl, 1 mM EDTA, pH 6.8. Pre-filter (0.22 µm), degas, and pre-chill if needed for sensitive proteins.

Operational tips:

• Prepare fresh buffer, pre-equilibrate temperature, and use immediately after adding the DMSO stock to limit maleimide hydrolysis.
• Desalt or buffer-exchange reduced proteins into amine-free buffer prior to conjugation.

Green Alternatives

Solvent and process choices dominate the environmental profile for handling Doxorubicin-SMCC. While the molecule itself is fixed, greener execution is possible (literature/general):

Greener solvent options and trade-offs:

• Aqueous buffers with minimal organic cosolvent: reduce VOC use; risk of maleimide hydrolysis increases above pH 7 and at longer times; solubility may limit concentration.
• Glycerol or PEG-400 as co-solvents: lower toxicity and volatility compared to DMF; viscosity and potential protein effects must be considered.
• Ethyl acetate or 2-MeTHF for workup/purification (if applicable to small-scale syntheses): greener than chlorinated solvents; ensure compound stability in these media.

Comparison (literature/general):

• DMSO: relatively benign EHS profile among polar aprotics; biodegradable; still requires proper waste handling.
• DMF: effective but reproductive toxin; prefer DMSO when feasible.

Process-intensity reductions:

• Perform reactions at ambient temperature and near-neutral pH to minimize energy input and byproduct formation.
• Use high-concentration stocks to reduce solvent volumes; adopt microscale conjugations to limit waste.
• Implement single-use, pre-aliquoted vials to reduce freeze–thaw degradation and associated waste.

Pharmaceutical Uses

No pharmacopeial or GMP status is claimed for this product. Item is supplied for research use only.

Formulation/processing roles in R&D (literature/general):

• Payload for constructing research-grade bioconjugates (e.g., protein, peptide, polymer, or nanoparticle conjugates) to study linker stability, drug–carrier ratios, and release mechanisms in vitro.
• Optical tracer: the inherent chromophore/fluorescence of doxorubicin facilitates in-process analytics (HPLC-UV/FLD) and qualitative visualization in formulation screens.
• Reference standard for method development: useful in establishing conjugation, purification, and analytical workflows for thiol–maleimide chemistry, including SEC, HIC, and LC–MS characterization of conjugates.

Important boundaries:

• Not intended for use as an API, excipient, or in any clinical manufacturing. No claims are made regarding sterility, endotoxin, residual solvent limits, or compendial compliance. Users must establish suitability and quality attributes for their specific research application.

Physical Properties

Item-specific physicochemical specifications are not provided for this SKU.

• 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 complex conjugates; typically decompose before boiling (literature/general).
• Density: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Not specified for this item; refer to CoA/Spec Sheet. General practice (literature): dissolve initially in anhydrous DMSO or DMF; for conjugations, dilute into buffered aqueous media (pH 6.5–7.0) immediately prior to use to minimize maleimide hydrolysis.
• LogP/logD: Not specified for this item; refer to CoA/Spec Sheet. The SMCC cyclohexyl spacer increases hydrophobic character relative to native doxorubicin (literature).
• pKa(s): Not specified for this item; refer to CoA/Spec Sheet. Anthracycline phenols and aminosugar display multiple ionizable sites (literature).
• UV/Vis: Not specified for this item; refer to CoA/Spec Sheet. Doxorubicin chromophore typically shows strong absorbance in the visible (~480–500 nm) and UV regions (literature), useful for tracking during conjugation.

Practical notes (literature/general):

• Maleimide moieties hydrolyze faster above pH ~7.5; keep solutions slightly acidic to neutral.
• Protect from light due to anthracycline chromophore photolability.

Quality and Grades

• Grade/Purity: Moligand™ (as listed). Item-specific assay/purity, residual solvents, and related substances are Not specified for this item; refer to CoA/Spec Sheet.

What Moligand™ implies (general description):

• The Moligand™ designation denotes inclusion in a small-molecule/ligand library suitable for discovery and conjugation workflows. Such materials are typically supplied to support screening, SAR, and bioconjugation method development in research settings. It does not, by itself, imply compliance with GMP or pharmacopeial monographs.

Analytical considerations for this class (literature/general):

• Identity/purity often assessed by LC–MS and HPLC with visible detection due to doxorubicin chromophore. Maleimide integrity can be monitored by derivatization with thiols and by 1H NMR (vinyl protons) or LC–MS mass shift.
• Low-water handling is critical; Karl Fischer moisture and maleimide content assays are commonly employed for quality control in bioconjugation-grade linkers.

Stabilizers and additives:

• Stabilizers, if any, are Not specified for this item; refer to CoA/Spec Sheet. Many maleimide reagents are shipped neat without stabilizers and retain activity when stored frozen and protected from moisture (literature).

Documentation:

• For batch-specific specifications (purity, counter-ions, form), consult the Certificate of Analysis and Specification Sheet prior to use.

Reaction and Applications

This reagent is designed for thiol-directed bioconjugation, leveraging the maleimide of the SMCC spacer to attach a doxorubicin payload to cysteine-containing targets (literature/general).

Typical applications:

• Site-selective conjugation to reduced cysteines on proteins, peptides, or engineered scaffolds via Michael addition to the maleimide, forming stable thioether linkages (literature).
• Preparation of doxorubicin-functionalized polymers, nanoparticles, and surfaces by thiol–maleimide coupling (materials and chemical biology research).
• Model systems for antibody–drug conjugate (ADC)-like constructs in discovery workflows; enables evaluation of linker–payload behavior (hydrophobicity, drug-to-carrier ratio) in vitro (research use only).

Mechanistic notes (literature):

• Maleimide reacts rapidly and selectively with thiolate anions at pH ~6.5–7.0, minimizing competing amine reactions and hydrolysis to maleamic acid.
• The SMCC cyclohexyl spacer enhances hydrophobicity and can influence conjugate stability and aggregation propensity; consider co-solvents and surfactants for protein work.

Practical guidance:

• Reduce disulfides with mild reagents (TCEP preferred over DTT to avoid thiol carryover), then desalt prior to conjugation.
• Control stoichiometry to target desired substitution (e.g., 1–4 equivalents per thiol), and quench residual maleimide with cysteine or mercaptoethanol after reaction.
• Monitor progress by UV–Vis (doxorubicin absorbance), LC–MS, or SDS-PAGE (for proteins).

Reaction Conditions

General, literature-based guidance for thiol–maleimide conjugation using Doxorubicin-SMCC; adjust to your system and consult primary literature.

• Solvent/buffer: 10–50 mM phosphate or HEPES buffer, pH 6.5–7.0, 50–150 mM NaCl, 1–5 mM EDTA; ≤10% DMSO as cosolvent to aid solubility.
• Temperature: 4–25°C. Lower temperatures preserve sensitive proteins; higher end accelerates reaction but may increase hydrolysis.
• Stoichiometry: 1.0–1.5 equivalents of maleimide per free thiol for selective labeling; up to 3–5 eq for higher loadings (proteins may require optimization).
• Time: 30 min to 2 h for peptides/proteins at µM–mM; monitor reaction to avoid over-conjugation/aggregation.
• Quench: add excess cysteine or 2-mercaptoethanol to consume residual maleimide after desired labeling.
• Reduction (pre-step): reduce disulfides with TCEP (0.5–5 mM) for 15–60 min at pH ~7; remove TCEP by desalting if it interferes with downstream analysis.
• Analysis: track by UV–Vis (doxorubicin Amax in visible range), LC–MS for small conjugates; for proteins, intact MS, SDS-PAGE fluorescence, and HIC/SEC to assess drug-to-carrier ratio and aggregation.

Notes:

• Avoid amine-containing buffers (Tris) and competing thiols (DTT) during the coupling step.
• Maleimide ring opening increases rapidly above pH 7.5, lowering thiol reactivity; keep pH controlled and reaction times modest.

Safety and Handling

Safety data specific to this item are not provided here; consult the SDS for authoritative guidance.

GHS classification (item-specific):

• Signal word: Not specified for this item; refer to SDS.
• H-statements: Not specified for this item; refer to SDS.
• Pictograms: Not specified for this item; refer to SDS.

General hazard information (literature/general):

• Doxorubicin derivatives are potent cytotoxic and genotoxic chemicals. Handle in a certified chemical fume hood with appropriate containment. Avoid inhalation, ingestion, skin/eye contact.
• Maleimide-functional compounds are electrophilic and can alkylate thiols; avoid contact with skin and reducing biological materials.

PPE and engineering controls:

• Wear lab coat, double nitrile gloves (change frequently), chemical splash goggles; use fume hood and, if available, a cytotoxic-drug handling workstation.

First aid (general):

• Skin/eye contact: Immediate decontamination with copious water for ≥15 min; remove contaminated clothing; seek medical evaluation.
• Inhalation: Move to fresh air; monitor respiration; seek medical attention.
• Ingestion: Rinse mouth; do not induce vomiting; obtain medical aid.

Storage incompatibilities and precautions (literature/general):

• Protect from light and moisture. Avoid prolonged exposure to aqueous buffers, bases, and nucleophiles which promote maleimide hydrolysis/ring opening.
• Segregate from oxidizers and reducing agents; store in tightly sealed, inert-atmosphere containers when possible.

Research use only. Not for human or veterinary use.

Solvent Selection

Doxorubicin-SMCC is typically handled as a concentrated stock in polar aprotic solvents, then diluted into buffers immediately prior to conjugation (literature/general).

• Primary stock solvents: anhydrous DMSO or DMF provide good solubility for anthracycline-linker conjugates and preserve maleimide functionality.
• Co-solvent strategy: for aqueous conjugations, pre-dissolve in DMSO (e.g., 10–20 mM), then add to buffer to a final organic content ≤10% v/v to limit precipitation and maleimide hydrolysis (literature guidance).
• Buffer compatibility: use mildly acidic to neutral buffers (pH 6.5–7.0) without competing nucleophiles; avoid primary amine buffers (e.g., Tris) that can consume maleimides (literature).

Comparison (literature/general):

• DMSO: highest solvating power, miscible with water; can scavenge radicals; widely used for bioconjugations.
• DMF: similar solvating power; less viscous; also miscible; ensure low water content.
• Aqueous-only: often limited by solubility and maleimide stability; feasible at low concentrations with appropriate pH control.

Practical tips:

• Filter stock solutions (0.2 µm PTFE) if particulates are present; avoid cellulose filters with strong organics.
• Use amber vials and dry inert gas headspace to limit oxidation and hydrolysis.

Storage and Reconstitution

Item-specific storage and shipping conditions:

• Storage: Store at -80°C (per Product Data). Keep tightly capped, desiccated, and protected from light.
• Shipping: Shipped on dry ice packs + cold packs (per Product Data).

Reconstitution and handling (literature/general):

• Solvents: Reconstitute in anhydrous DMSO or DMF to prepare concentrated stocks (e.g., 10–20 mM). Vortex gently and, if needed, sonicate briefly to aid dissolution.
• Aliquoting: Dispense single-use aliquots into dry, amber vials or tubes under inert gas to minimize moisture/oxygen exposure. Avoid repeated freeze–thaw cycles.
• Working solutions: Dilute stocks into conjugation buffer immediately before use; discard any buffer-diluted solutions after the experiment due to maleimide hydrolysis risk.
• Light protection: Use amber containers and minimize ambient light exposure to preserve the anthracycline chromophore.

Stability:

• Long-term stability, shelf-life, and permissible freeze–thaw cycles are Not specified for this item; refer to CoA/Spec Sheet. As a general practice, maintain continuous frozen storage and avoid prolonged exposure to aqueous conditions.

Disposal: Treat wastes as cytotoxic and chemically hazardous; follow institutional and regulatory procedures.

Structure and Identity

This item is a doxorubicin derivative bearing an SMCC spacer intended for thiol-directed bioconjugation (literature). Doxorubicin-SMCC typically comprises the anthracycline aglycone/sugar of doxorubicin covalently linked through an amide to the cyclohexyl-based SMCC linker, leaving a terminal maleimide for conjugation to sulfhydryl groups (literature). Item-specific identifiers are not provided for this SKU.

• SKU: D1494975
• Product name: Doxorubicin-SMCC
• CAS: 400647-59-8
• 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 (literature/general):

• Core motifs: anthracycline tetracyclic ring system with daunosamine sugar (from doxorubicin); cyclohexane-1-carboxamide spacer from SMCC; maleimide electrophile for thiol coupling.
• Functional groups: multiple phenolic hydroxyls, ketone, amino sugar (often acylated), amide linkage(s), maleimide (Michael acceptor), and hydrophobic cyclohexyl moiety.
• 2D description: polycyclic aromatic chromophore (planar) fused to a glycosidic aminosugar; via an amide tether to a cyclohexane ring bearing a methylene–maleimide substituent.

Notes:

• Exact regiochemistry and substitution pattern may vary by synthesis route; consult the CoA/SDS for definitive structure for this catalog item.

Synthetic Utility

From a synthetic and bioconjugation perspective (literature/general):

• Functional handle: a terminal maleimide enables chemoselective coupling to thiols on biomolecules, small molecules, and materials, forming robust thioether bonds under mild aqueous conditions.
• Spacer design: the SMCC cyclohexyl unit provides a non-cleavable, hydrophobic spacer that can modulate distance and conformational freedom between doxorubicin and the target. Non-cleavable linkers are valued for stability studies and as controls versus cleavable linkers.
• Orthogonality: by pre-installing SMCC on doxorubicin (consuming the NHS ester end), the remaining functionality is orthogonal to amine chemistry, allowing sequential strategies (e.g., first label lysines with azides, then introduce doxorubicin via thiol–maleimide reaction).
• Retrosynthetic perspective: Doxorubicin-SMCC can be viewed as a payload module in modular assembly of conjugates; downstream diversification occurs at the maleimide through thiol-bearing partners (proteins, polymers, small molecules).

Use cases:

• Constructing standards to benchmark alternative linkers (e.g., maleimide vs. haloacetamide or click-chemistry linkers).
• Surface immobilization of doxorubicin onto thiolated self-assembled monolayers or resins for affinity or mechanistic studies.

Target Specificity

This product is a small-molecule linker–payload and does not have inherent immunological target specificity like an antibody or affinity reagent.

• Antigen/epitope: Not applicable.
• Species reactivity: Not applicable.
• Clone/isotype: Not applicable.

For selective conjugation, specificity arises from the chemistry: the terminal maleimide reacts preferentially with thiols over other nucleophiles at pH ~6.5–7.0 (literature/general). Any biological targeting must be provided by the chosen carrier (e.g., antibody, peptide, nanoparticle) rather than by Doxorubicin-SMCC itself.