Product Description

1、Norcantharidin (CAS 29745-04-8) is a versatile, research-grade, high-purity small molecule derived from cantharidin. Its chemical structure enables potent inhibition of protein phosphatases, particularly PP1 and PP2A, which are critical regulators of cell cycle progression, apoptosis, and intracellular signaling cascades. By modulating these phosphatases, Norcantharidin induces accumulation of phosphorylated proteins, leading to alterations in cellular homeostasis, tumor growth suppression, and apoptosis induction in in vitro experimental models.

This compound has been extensively utilized in oncology research to investigate tumor cell proliferation, metastasis, and molecular signaling networks. Norcantharidin’s ability to induce G2/M phase arrest makes it a valuable tool for studying mitotic checkpoint regulation and DNA damage response mechanisms in a controlled laboratory setting. Researchers employ Norcantharidin to examine how inhibition of phosphatase activity influences downstream pathways such as MAPK/ERK, AKT, NF-κB, and p53, which are critical for tumor cell survival, stress response, and chemoresistance modeling.

The high-purity bulk format ensures consistent experimental results, allowing laboratories to perform dose-response studies, high-throughput screenings, and mechanistic pathway investigations. Its physicochemical properties, including crystalline powder stability and solubility in DMSO and other organic solvents, facilitate precise dosing and reproducibility across diverse experimental setups.

Norcantharidin’s applications extend to studying molecular mechanisms underlying tumor progression and chemotherapeutic resistance. By using this compound, researchers can simulate intracellular signaling disruptions that mimic the effects of therapeutic phosphatase inhibition, enabling investigation into compensatory pathway activation and adaptive cellular responses. Additionally, Norcantharidin serves as a reference inhibitor in protein phosphatase research, providing a benchmark for screening novel small-molecule modulators of PP1 and PP2A.

From a mechanistic perspective, Norcantharidin interacts with key residues in the catalytic sites of PP1 and PP2A, forming stable enzyme-inhibitor complexes. These interactions block the dephosphorylation of downstream targets, resulting in enhanced phosphorylation levels of cell cycle regulators, apoptotic mediators, and stress-responsive proteins. The ability to modulate phosphorylation dynamics makes Norcantharidin an indispensable reagent for research on cellular signal transduction, DNA damage responses, and programmed cell death pathways.

In addition to classical cancer cell line studies, Norcantharidin is employed in organoid cultures, three-dimensional tumor spheroids, and ex vivo tissue models. This versatility allows researchers to explore complex microenvironment interactions, tumor-stroma signaling, and heterogeneity in kinase and phosphatase activity. Its reproducibility and high-purity quality make it ideal for comparative studies across multiple laboratory settings, ensuring consistency in experimental outputs.

The compound is produced under GMP-compliant conditions with stringent quality control procedures to ensure batch-to-batch consistency. Analytical methods including HPLC, LC-MS/MS, and NMR spectroscopy confirm purity ≥99% and absence of significant impurities. Such quality assurance is critical for laboratories conducting sensitive mechanistic studies, kinase profiling, or apoptosis assays where trace contaminants could confound results.

2、Norcantharidin also serves as a model compound for exploring mechanisms of chemoresistance in cancer cells. By inhibiting PP1 and PP2A, it reveals how cells compensate for loss of phosphatase activity through upregulation of alternative signaling pathways such as PI3K/AKT, MAPK/ERK, and NF-κB. Researchers can leverage these responses to investigate potential combination therapies, validate molecular targets, and map adaptive survival pathways in tumor models.

The compound’s crystalline powder is highly amenable to controlled solubilization, ensuring reproducible dosing in DMSO, ethanol, methanol, or buffered aqueous media. This feature allows laboratories to perform time-course experiments, dose-response analyses, and mechanistic pathway inhibition studies with minimal variability. Its stability under refrigerated conditions (2–8°C) further ensures long-term storage without significant degradation, preserving experimental fidelity.

Norcantharidin has been incorporated in numerous laboratory protocols including:

• Apoptosis induction and caspase activation studies

• G2/M cell cycle checkpoint investigations

• Tumor spheroid and 3D culture proliferation assays

• Phosphoproteomics and signaling pathway profiling

• DNA damage and repair mechanistic analysis

• Drug-resistance modeling in ALK, EGFR, and MAPK-driven cells

In these studies, the compound demonstrates high selectivity, allowing researchers to focus on PP1/PP2A-mediated pathways without substantial off-target interference. Its high-purity specification ensures that observed cellular responses are attributable to the compound itself rather than contaminants, providing confidence in experimental reproducibility.

The bulk and GMP-compliant format of Norcantharidin makes it ideal for multi-lab collaborative research projects, high-throughput screenings, and long-term mechanistic studies. Its consistent physicochemical properties enable seamless integration into automated laboratory systems, robotic pipetting workflows, and microfluidic assay platforms.

By providing a reliable, high-purity research chemical, Norcantharidin allows scientists to investigate the fundamental roles of phosphatase inhibition in cancer biology, uncover novel molecular targets, and generate high-fidelity preclinical data.

Product Specifications

The following specifications provide a comprehensive overview of Norcantharidin’s analytical properties, structural characteristics, stability, and laboratory suitability.

Product Specification Table

Expanded Explanation of Specifications

Norcantharidin’s chemical integrity is confirmed through rigorous analytical testing including HPLC, LC-MS/MS, and NMR spectroscopy. These analyses verify the compound’s purity (≥99%), confirm its molecular structure, and ensure minimal impurities that could interfere with mechanistic studies.

The bulk crystalline powder form offers excellent stability and ease of handling in laboratory conditions. Solubility in DMSO, methanol, and ethanol ensures rapid preparation of working solutions, facilitating integration into cellular assays, enzyme inhibition studies, and multi-well plate experiments. Its slightly limited water solubility can be addressed by solvent dilution or pH-buffered solutions depending on experimental needs.

Batch consistency is maintained under GMP-aligned production, providing minimal variation between lots. This ensures reproducible results in longitudinal studies, cross-laboratory collaborations, and high-throughput screening. Endotoxin testing further ensures compatibility with sensitive cell culture systems, reducing confounding immune responses during in vitro experiments.

Norcantharidin’s robust physicochemical and analytical profile makes it an indispensable tool for studies of phosphatase-mediated cellular regulation, apoptosis induction, cell cycle modulation, and tumor biology research.

Mechanism of Action

Norcantharidin (CAS 29745-04-8) functions primarily as a potent protein phosphatase inhibitor, selectively targeting PP1 and PP2A. These phosphatases regulate phosphorylation homeostasis of numerous signaling proteins, including those involved in cell cycle progression, apoptosis, DNA repair, and stress response pathways. Inhibition of PP1/PP2A by Norcantharidin leads to accumulation of phosphorylated substrates, resulting in multiple downstream effects in cancer and normal cell models used in laboratory research.

Phosphatase Inhibition Dynamics

PP1 and PP2A are serine/threonine phosphatases that control critical cellular processes. By binding to their catalytic domains, Norcantharidin prevents dephosphorylation of substrates such as cyclin-dependent kinases, checkpoint regulators, and transcription factors. This enzymatic blockade induces:

• Hyperphosphorylation of cell cycle proteins (e.g., cyclin B1, CDK1) leading to G2/M arrest

• Activation of apoptosis mediators (e.g., Bax, cleaved caspases)

• Modulation of survival pathways (AKT, ERK, NF-κB) to study adaptive signaling mechanisms

The inhibitory kinetics are concentration- and time-dependent, allowing laboratories to map dose-response relationships, enzymatic inhibition curves, and temporal changes in phosphorylation states.

Apoptosis Induction Mechanism

Norcantharidin induces apoptosis through both intrinsic and extrinsic pathways:

• Mitochondrial pathway: Increases Bax/Bcl-2 ratio, triggers mitochondrial membrane depolarization, releases cytochrome c, and activates caspase-9 and caspase-3

• Death receptor pathway: Modulates Fas/FasL expression in sensitive tumor models

• ROS-mediated stress: Elevates reactive oxygen species, promoting apoptosis in selected cell lines

These effects are widely utilized in laboratory assays to study programmed cell death, chemoresistance, and combination therapy strategies.

Cell Cycle Arrest

By hyperphosphorylating checkpoint regulators, Norcantharidin induces G2/M arrest, a critical tool in research on mitotic checkpoints. Researchers use this feature to:

• Investigate DNA damage responses

• Model mitotic catastrophe

• Explore synergy with other chemotherapeutic agents

• Examine phosphatase-dependent regulation of cyclins and CDKs

Signal Transduction Modulation

Norcantharidin’s inhibition of PP1/PP2A affects multiple intracellular pathways:

• MAPK/ERK pathway: Downregulation leads to decreased proliferation

• PI3K/AKT pathway: Inhibition reduces survival signals and sensitizes cells to apoptosis

• NF-κB pathway: Suppressed transcriptional activity reduces anti-apoptotic gene expression

This multifaceted modulation allows laboratories to explore network-level signaling and compensatory mechanisms under phosphatase inhibition.

DNA Damage and Repair Studies

Hyperphosphorylation of H2AX (γ-H2AX) occurs after Norcantharidin treatment, marking DNA double-strand breaks. Researchers employ this property to study:

• DNA damage sensing and repair kinetics

• Genotoxic stress responses

• Tumor resistance mechanisms in cancer models

These applications are critical for understanding therapeutic vulnerabilities and designing experimental strategies for drug development.

Applications in Resistance Modeling

Chronic exposure studies using Norcantharidin allow exploration of:

• Adaptive cellular resistance pathways

• Crosstalk between kinase and phosphatase networks

• Biomarker discovery for chemotherapeutic sensitivity

These experiments enable laboratories to map survival signaling, identify novel targets, and simulate clinically relevant resistance scenarios.

Experimental Advantages of Bulk High-Purity Compound

Norcantharidin’s GMP-compliant bulk production ensures:

• Batch consistency for longitudinal studies

• High reproducibility across labs

• Solvent compatibility with DMSO, ethanol, methanol, and buffered media

• Integration into automated assays for high-throughput screening

Its reliable physicochemical profile allows precise control over concentration, exposure duration, and experimental reproducibility, which is essential for mechanistic studies.

Side Effects

Although Norcantharidin is intended for research use only, its effects on cells and molecular systems have been extensively characterized in vitro. These effects provide insight into its pharmacological mechanisms and are important for laboratory safety.

Cellular Effects

• Cytotoxicity: Dose-dependent reduction in proliferation of tumor cell lines

• Apoptotic features: Nuclear condensation, DNA fragmentation, membrane blebbing

• Morphological changes: Cell shrinkage, vacuolation, and detachment from culture surfaces

• Stress responses: Mitochondrial depolarization and ROS generation

Molecular Effects

• Hyperphosphorylation of substrates due to PP1/PP2A inhibition

• Activation of pro-apoptotic mediators and suppression of anti-apoptotic proteins

• Signal pathway modulation: Decreased AKT, ERK, and NF-κB activity, leading to apoptosis and cell cycle arrest

• DNA damage signaling: Increased γ-H2AX expression reflecting double-strand break accumulation

Laboratory Handling Considerations

• Personal Protective Equipment: Gloves, lab coats, eye protection recommended

• Aerosol Prevention: Handle powders in fume hoods or biosafety cabinets

• Avoid Ingestion/Inhalation: Highly concentrated powders may cause cellular toxicity

• Storage: 2–8°C in sealed containers, protected from light

• Waste Disposal: Follow institutional chemical waste protocols

Norcantharidin’s sterile bulk format reduces particulate contamination, ensures accurate dosing, and improves reproducibility compared to non-standardized preparations.