Doxorubicin

Synthetic Routes and Reaction Conditions: Doxorubicin is synthesized from daunorubicin, another anthracycline antibiotic. The synthesis involves several steps, including glycosylation, oxidation, and methylation. The key intermediate in the synthesis is daunorubicin, which undergoes hydroxylation to form this compound .

Industrial Production Methods: Industrial production of this compound typically involves fermentation using the bacterium Streptomyces peucetius. The fermentation process is followed by extraction and purification steps to isolate this compound. Advanced techniques such as remote loading and PEGylation are used to enhance the stability and efficacy of this compound liposomes .

DNA Intercalation and Structural Interactions

Doxorubicin intercalates into DNA through its planar anthraquinone ring, forming hydrogen bonds with nucleotide bases. This interaction introduces torsional stress, destabilizes nucleosome structures, and inhibits topoisomerase II activity .

Key Features :

• Base Pair Preference : Demonstrates higher affinity for GC-rich regions due to increased hydrogen bonding (4–5 direct bonds in GC vs. 3–4 in AT sequences) .

• Helix Distortion : Molecular dynamics show C2′-endo pucker in DNA, widening the major groove and exposing the D-ring of this compound to solvent .

Table 1: DNA-Doxorubicin Interaction Parameters

Redox Reactions and Reactive Oxygen Species (ROS) Generation

This compound undergoes redox cycling via two pathways:

• Two-Electron Reduction : Catalyzed by carbonyl reductases (CBR1, AKR1C3), forming doxorubicinol (C₂₇H₃₁NO₁₁), a less active metabolite .

• One-Electron Reduction : Generates a semiquinone radical (C₂₇H₂₈NO₁₁⁻), which reacts with O₂ to produce superoxide (O₂⁻) and hydrogen peroxide (H₂O₂) .

Critical Implications :

• Cardiotoxicity : ROS from one-electron reduction cause lipid peroxidation and mitochondrial damage .

• pH Sensitivity : Redox activity increases at lower pH (e.g., tumor microenvironments) .

Dimerization and Precipitation

In neutral buffers (e.g., PBS), this compound forms covalent dimers via imine linkages between C-13/C-14 keto groups and amino sugars .

Key Observations :

• Dimer Structure : Proposed as a C-13/C-14 crosslinked product (C₅₄H₅₆N₂O₂₂) with m/z 1069.2 .

• Kinetics : Follows pseudo-first-order kinetics with a rate constant (k) of 0.12 h⁻¹ at pH 7.4 .

Table 2: Dimerization Conditions and Outcomes

Degradation Pathways

This compound degrades under acidic and alkaline conditions:

• Acidic Hydrolysis (pH <4) : Cleavage of glycosidic bond, yielding doxorubicinone (aglycone, C₂₁H₁₈O₈) and daunosamine .

• Alkaline Hydrolysis (pH >8) : Oxidation of the C-9 hydroxyl group to a ketone, forming 7-deoxydoxorubicinolone .

Stability Data :

• Half-Life : 48 h (pH 7.4, 25°C) vs. 12 h (pH 9.0, 37°C) .

• Buffer Catalysis : Phosphate and acetate buffers accelerate degradation by 2–3× .

Metabolic Transformations

Major Pathways :

• Hydroxylation : CYP3A4/5-mediated formation of doxorubicinol (reduced cardiotoxicity) .

• Deglycosidation : Loss of daunosamine sugar, yielding inactive metabolites.

Table 3: Metabolic Enzyme Contributions

Synthetic Modifications

Stereochemical alterations to the daunosamine sugar (e.g., N,N-dimethylepirubicin) reduce DNA intercalation while retaining cytotoxicity .

Formulation Advancements

Recent advancements in drug formulation have significantly improved the delivery and efficacy of doxorubicin. Various formulations have been developed to enhance its therapeutic index while minimizing side effects:

These formulations leverage nanotechnology and targeted delivery systems to improve drug accumulation in tumor tissues while reducing systemic toxicity.

Clinical Case Studies

Case Study 1: Breast Cancer Treatment
In a study evaluating this compound's effectiveness in combination with cyclophosphamide for stage II to III triple-negative breast cancer, the regimen showed a significant increase in pathologic complete response rates compared to other treatments .

Case Study 2: Ovarian Cancer
Liposomal this compound has demonstrated efficacy in patients with recurrent ovarian cancer who had previously undergone platinum-based therapies. The liposomal formulation allowed for higher concentrations of the drug to accumulate in tumor tissues with reduced side effects .

Case Study 3: Glioblastoma Therapy
The use of this compound-loaded iron oxide nanoparticles was explored for glioblastoma treatment. Results indicated substantial tumor reduction and increased survival rates due to targeted delivery mechanisms that enhanced drug penetration through the blood-brain barrier .

Mechanisms Overcoming Resistance

This compound's effectiveness can be hampered by multidrug resistance (MDR) mechanisms. Recent studies have focused on strategies to overcome this challenge:

• Targeted Delivery Systems : Immunoliposomal formulations have been shown to enhance drug uptake in resistant cancer cells by targeting specific receptors like EGFR .
• Combination Therapies : Combining this compound with other agents (e.g., iron chelators) can mitigate oxidative damage and enhance therapeutic outcomes .

Doxorubicin exerts its effects by intercalating with DNA, thereby inhibiting the action of topoisomerase II, an enzyme involved in DNA replication. This leads to the generation of reactive oxygen species and subsequent DNA damage, ultimately resulting in cell death . This compound also affects various molecular targets and pathways, including the Bcl-2/Bax apoptosis pathway, which plays a crucial role in regulating cell death .

• Daunorubicin
• Epirubicin
• Idarubicin
• Mitoxantrone

Comparison: Doxorubicin is unique among these compounds due to its broad spectrum of activity against various cancers and its ability to intercalate with DNA. While daunorubicin is structurally similar to this compound, it has a different side chain, which affects its pharmacokinetic properties . Epirubicin and idarubicin are also anthracyclines but have different toxicity profiles and clinical applications . Mitoxantrone, although not an anthracycline, shares some structural similarities and is used in similar clinical settings .

Doxorubicin, an anthracycline antibiotic, is widely utilized in chemotherapy for various cancers, including breast cancer, leukemia, and lymphomas. Its effectiveness is attributed to its ability to intercalate DNA, thereby inhibiting topoisomerase II and disrupting the replication of cancer cells. However, its clinical use is often limited by dose-dependent cardiotoxicity. This article delves into the biological activities of this compound, examining its mechanisms of action, therapeutic efficacy, associated risks, and recent research findings.

This compound exerts its cytotoxic effects primarily through:

• DNA Intercalation : this compound intercalates between DNA base pairs, leading to structural distortions that inhibit DNA replication and transcription.
• Topoisomerase II Inhibition : It stabilizes the topoisomerase II-DNA complex, preventing DNA strand re-ligation and resulting in double-strand breaks.
• Reactive Oxygen Species (ROS) Generation : this compound induces oxidative stress through ROS production, leading to apoptosis in cancer cells .

Efficacy in Cancer Treatment

This compound has demonstrated significant efficacy across various malignancies. A summary of its application in different cancers is presented below:

Case Studies

• This compound-Induced Cardiomyopathy :
  A 68-year-old female with pleural epithelioid angiosarcoma developed heart failure following this compound treatment. Initial heart function was assessed using left ventricular ejection fraction (LVEF), which deteriorated post-treatment. After implementing cardioprotective strategies, including sacubitril/valsartan, her heart function improved over time .
• Long-term Effects :
  A study reported a 57-year-old woman who developed acute shortness of breath due to this compound-induced cardiomyopathy 17 years after chemotherapy for breast cancer. This case highlights the delayed onset of cardiotoxic effects associated with this compound .

Recent Research Findings

Recent studies have focused on mitigating the cardiotoxic effects while maintaining the anti-cancer efficacy of this compound:

• BAI1 and BAX Interaction : Research indicates that targeting BAX, a pro-apoptotic protein activated by this compound, may prevent cardiac cell death without compromising anti-tumor activity. This dual approach could enhance the therapeutic window of this compound .
• Transcriptional Changes : Exposure to this compound induces significant transcriptional changes in cellular metabolism and stress response pathways. Studies using Saccharomyces cerevisiae have shown upregulation of genes involved in oxidative stress response and DNA repair mechanisms .

Basic Research Questions

Q. What experimental models are considered gold standards for evaluating doxorubicin’s antitumor efficacy and cardiotoxicity?

• Methodological Answer : Utilize in vivo rodent models (e.g., xenograft tumors) to assess tumor regression, paired with echocardiography or histopathology to quantify cardiac damage. Biomarkers like troponin and BNP are critical for early detection of cardiotoxicity . For in vitro studies, human cardiomyocyte cell lines (e.g., AC16) and cancer cell lines (e.g., A549) are standardized models. Ensure dose ranges align with clinically relevant plasma concentrations (e.g., 0.1–5 µM) .

Q. How should researchers design studies to validate this compound’s pharmacokinetic properties and formulation stability?

• Methodological Answer : Employ high-performance liquid chromatography (HPLC) to measure plasma half-life and tissue distribution. Preformulation studies should evaluate interactions with excipients or proteins (e.g., bovine serum albumin) using spectroscopic methods (UV-Vis, fluorescence) under varying pH and temperature conditions . Stability testing must follow ICH guidelines, monitoring degradation products under accelerated conditions .

Q. What are the established methodologies for assessing cumulative dose-dependent cardiotoxicity in longitudinal studies?

• Methodological Answer : Retrospective cohort analyses of clinical data (e.g., cumulative doses >450 mg/m²) remain foundational. Preclinically, use serial echocardiography in murine models, with endpoints like left ventricular ejection fraction (LVEF) decline. Stratify risk by age and dosing schedules (e.g., weekly vs. triweekly administration) .

Advanced Research Questions

Q. How can researchers address heterogeneity in preclinical meta-analyses of this compound’s efficacy and toxicity?

• Methodological Answer : Mitigate heterogeneity via subgroup analyses (e.g., exercise protocols, dosing intervals) and strict inclusion criteria during systematic reviews. Use tools like I² statistics to quantify variability. Prioritize studies with standardized endpoints (e.g., tumor volume reduction ≥50%) and transparent reporting of negative results to reduce publication bias .

Q. What experimental designs optimize the evaluation of this compound combination therapies (e.g., with kinase inhibitors or immunotherapies)?

• Methodological Answer : Implement randomized phase II trials with stratification by tumor type, performance status, and prior therapies. For preclinical studies, use syngeneic models to assess immune modulation. Measure synergistic effects via Chou-Talalay combination indices and validate using transcriptomic profiling (e.g., RNA-seq for apoptosis pathways) .

Q. How can transcatheter intraarterial delivery methods improve this compound’s tumor targeting in hepatocellular carcinoma?

• Methodological Answer : Compare drug distribution using fluorescence microscopy in VX2 rabbit liver tumor models. Quantify intratumoral this compound concentration via mass spectrometry and correlate with tumor necrosis rates. Optimize catheterization techniques (e.g., flow rate, embolization agents) to enhance localized exposure .

Q. What strategies validate preclinical findings on this compound’s cardiotoxicity mechanisms for clinical translation?

• Methodological Answer : Integrate multi-omics data (proteomics, metabolomics) from preclinical models with patient serum biomarkers. Validate mitochondrial dysfunction pathways (e.g., ROS production) using human induced pluripotent stem cell-derived cardiomyocytes. Conduct pilot clinical trials with cardiac MRI for subclinical toxicity detection .

Q. How do protein-binding interactions influence this compound’s stability and bioavailability in novel formulations?

• Methodological Answer : Use experimental design (e.g., Box-Behnken) to test this compound-BSA binding affinity under varying molar ratios. Characterize complexes via dynamic light scattering (DLS) and circular dichroism. Correlate binding constants (Ka) with in vivo bioavailability using pharmacokinetic modeling .

Q. What statistical methods are robust for analyzing time-to-event data in this compound clinical trials with high dropout rates?

• Methodological Answer : Apply Kaplan-Meier survival analysis with log-rank tests for between-group comparisons. Address censored data using Cox proportional hazards models, adjusting for covariates like age and cumulative dose. Validate assumptions with Schoenfeld residuals .