3-(4-Nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione

The synthesis of 3-(4-Nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione involves several steps. One common method includes the reaction of methyl 2-(bromomethyl)-3-nitrobenzoate with 3-aminopiperidine-2,6-dione hydrochloride in the presence of triethylamine and dimethyl sulfoxide . The reaction mixture is heated to 50-55°C, and the product is purified to obtain the desired compound .

Industrial production methods often involve solvent recrystallization, desolvation, vapor diffusion, and rapid precipitation techniques to obtain polymorphs of the compound .

3-(4-Nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione undergoes various chemical reactions, including:

Oxidation: The nitro group can be reduced to an amino group under specific conditions.

Reduction: The compound can be reduced to form 3-(4-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione.

Substitution: The compound can undergo nucleophilic substitution reactions, where the nitro group is replaced by other functional groups.

Common reagents used in these reactions include reducing agents like hydrogen gas and catalysts such as palladium on carbon . The major products formed from these reactions include 3-(4-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione and other substituted derivatives .

Chemical Properties and Structure

The compound has the molecular formula $C_{13}H_{11}N_{3}O_{5}$ and a molecular weight of 289.24 g/mol. Its structure features a piperidine ring substituted with a nitroisoindolinone moiety, which is crucial for its biological activity and interaction with various biological targets.

Anticancer Activity

Research indicates that 3-(4-Nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione exhibits significant anticancer properties. It is being investigated for its ability to inhibit tumor growth and metastasis in various cancer models. The compound's mechanism of action may involve modulation of cellular pathways associated with apoptosis and cell cycle regulation.

Anti-inflammatory Effects

The compound is also being explored for its anti-inflammatory properties. Studies suggest that it may inhibit pro-inflammatory cytokines and pathways involved in chronic inflammatory diseases. This application is particularly relevant for conditions such as rheumatoid arthritis and inflammatory bowel disease.

Polymorphism

The solid-state properties of this compound are critical for its formulation as a pharmaceutical agent. Different polymorphic forms have been identified, which can influence solubility, stability, and bioavailability. For instance:

Understanding these polymorphs allows researchers to optimize drug delivery systems tailored to patient needs.

Drug Delivery Systems

The compound's integration into drug delivery systems is under investigation. Its properties enable the development of novel formulations that enhance therapeutic efficacy while reducing side effects.

Mechanistic Studies

Biological studies are ongoing to elucidate the precise mechanisms through which this compound exerts its effects on cellular systems. These studies focus on:

• Interaction with specific protein targets
• Impact on signaling pathways
• Modulation of gene expression profiles

Preclinical Trials

Preclinical trials are being conducted to evaluate the safety and efficacy of this compound in vivo. Early results suggest promising outcomes in animal models for both cancer and inflammatory diseases.

The mechanism of action of 3-(4-Nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione involves its interaction with molecular targets and pathways. As an analog of Lenalidomide, it is believed to modulate the immune system, inhibit angiogenesis, and induce apoptosis in cancer cells . The compound binds to the ubiquitin E3 ligase cereblon, leading to the degradation of Ikaros transcription factors, which are crucial for the survival of multiple myeloma cells .

Structural Analogues

Key structural analogues of 3-(4-nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione include:

Physicochemical Properties

• Lenalidomide’s amino group enhances hydrogen-bonding capacity, improving bioavailability .
• HPLC Behavior : In stability-indicating HPLC methods, the nitro derivative (Impurity-A) elutes earlier than lenalidomide due to reduced polarity .

3-(4-Nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione, also known as 4-Nitro Lenalidomide, is a compound with significant biological activity, particularly in the context of cancer treatment and modulation of immune responses. This article explores its biological properties, mechanisms of action, and potential therapeutic applications.

• Molecular Formula : C$_{13}$H$_{11}$N$_{3}$O$_{5}$
• Molecular Weight : 289.24 g/mol
• CAS Number : 827026-45-9
• Physical State : White crystalline solid
• Melting Point : >250°C (dec.) .

The compound exhibits its biological activity primarily through its interaction with the IKAROS family zinc finger proteins (IKZF1, IKZF2, IKZF3, and IKZF4). These proteins are critical regulators of immune cell function and development. Specifically, this compound has been shown to:

• Degrade IKZF2 Protein : The compound selectively reduces IKZF2 levels, which is associated with enhanced regulatory T-cell (Treg) function and suppression of autoimmune responses. This activity suggests potential applications in treating autoimmune diseases and improving Treg-mediated immune regulation .
• Modulate Cytokine Production : It has been implicated in reducing levels of tumor necrosis factor (TNF), a cytokine involved in systemic inflammation. By modulating TNF levels, the compound may alleviate inflammatory conditions .

Cancer Treatment

Research indicates that this compound may have therapeutic potential in various cancers due to its ability to modulate immune responses and decrease tumor-promoting cytokines. For instance:

• Multiple Myeloma : In studies involving multiple myeloma cells, the compound demonstrated cytotoxic effects and enhanced apoptosis when combined with other therapeutic agents. This suggests a synergistic effect that could improve treatment outcomes .
• Autoimmune Diseases : The selective degradation of IKZF2 has been linked to improved Treg functionality in preclinical models of autoimmune disorders, indicating that this compound could serve as a novel treatment strategy for conditions like lupus or rheumatoid arthritis .

Data Table: Summary of Biological Activities

Q. Basic: What synthetic methodologies are recommended for preparing 3-(4-Nitro-1-oxoisoindolin-2-yl)piperidine-2,6-dione, and how can reaction yields be optimized?

Answer:
The synthesis typically involves multi-step reactions, including cyclization and nitro-group introduction. Key steps may mirror protocols for analogous isoindolinone-piperidine derivatives, such as using dichloromethane as a solvent and sodium hydroxide for deprotonation (as seen in nitro-substituted heterocycles) . To optimize yields:

• Temperature control : Maintain low temperatures during nitro-group incorporation to minimize side reactions.
• Catalyst selection : Use palladium or copper catalysts for efficient coupling reactions.
• Purification : Employ column chromatography with gradients (e.g., ethyl acetate/hexane) to isolate high-purity product (≥97%) .
  Computational reaction path searches (e.g., quantum chemical calculations) can predict optimal conditions, reducing trial-and-error approaches .

Q. Advanced: How does the nitro group at the 4-position influence the compound’s electronic properties and binding interactions in biological systems?

Answer:
The nitro group is electron-withdrawing, altering the isoindolinone ring’s electron density and potentially enhancing binding to target proteins (e.g., via π-π stacking or hydrogen bonding). Methodological approaches include:

• Spectroscopic analysis : IR and UV-Vis spectroscopy to study electronic transitions .
• Computational docking : Density Functional Theory (DFT) calculations to map electrostatic potential surfaces and predict binding affinities .
• Comparative studies : Substitute the nitro group with amino or fluorine moieties (as in related compounds) to assess structure-activity relationships (SAR) .

Q. Advanced: How can researchers resolve contradictions between computational predictions and experimental data on the compound’s stability?

Answer:
Contradictions often arise from solvent effects or unaccounted intermediates. A feedback loop integrating:

• Reaction path simulations : Use quantum chemical calculations (e.g., transition state analysis) to identify overlooked intermediates .
• In-situ monitoring : Employ techniques like NMR or HPLC-MS to track degradation products under varying conditions (pH, temperature) .
• Thermogravimetric analysis (TGA) : Quantify thermal stability discrepancies between predicted and observed data .

Q. Basic: What analytical techniques are essential for characterizing the compound’s structural integrity and purity?

Answer:
Critical techniques include:

• NMR spectroscopy : Confirm regiochemistry (e.g., nitro group position) via $^1$H and $^13$C chemical shifts .
• High-resolution mass spectrometry (HRMS) : Validate molecular weight (e.g., C$_{13}$H$_{12}$N$_3$O$_5$ requires m/z ~314.0778) .
• X-ray diffraction (XRD) : Resolve polymorphic forms (if present) and confirm crystal structure .
• HPLC : Assess purity (>95%) using reverse-phase columns and UV detection .

Q. Advanced: What experimental strategies are effective for identifying and characterizing polymorphic forms of this compound?

Answer:
Polymorph screening involves:

• Solvent recrystallization : Test polar (DMSO) vs. non-polar (toluene) solvents to induce different crystal forms .
• Differential scanning calorimetry (DSC) : Detect melting point variations between polymorphs.
• Dynamic vapor sorption (DVS) : Assess hygroscopicity and stability under humidity .
• Powder XRD : Compare diffraction patterns to reference data for known polymorphs .

Q. Basic: What storage conditions are critical to maintaining the compound’s stability during long-term studies?

Answer:

• Temperature : Store at 2–8°C to prevent thermal degradation .
• Atmosphere : Use inert gas (N$_2$ or Ar) in sealed vials to avoid oxidation .
• Light protection : Amber glass containers to block UV-induced nitro-group photoreduction .

Q. Advanced: How can the piperidine-2,6-dione moiety’s conformational flexibility impact the compound’s pharmacological activity?

Answer:
The piperidine ring’s puckering affects binding to enzymes like proteasomes or kinases. Methods to study this include:

• Molecular dynamics (MD) simulations : Model ring flexibility and predict bioactive conformers .
• Nuclear Overhauser effect (NOE) NMR : Measure interproton distances to map solution-state conformations .
• SAR studies : Compare activity of rigid analogs (e.g., spirocyclic derivatives) to assess flexibility-activity relationships .

Q. Advanced: How can quantum chemical calculations guide the design of novel derivatives with enhanced bioactivity?

Answer:

• Frontier molecular orbital (FMO) analysis : Identify reactive sites for functionalization (e.g., nitro reduction to amino groups) .
• Docking studies : Screen virtual libraries against target proteins (e.g., cereblon for immunomodulatory effects) .
• Free-energy perturbation (FEP) : Predict binding affinity changes for proposed derivatives .