Nilotinib

Nilotinib can be synthesized through various methods. One common synthetic route involves the conversion of a compound of formula (IV) into the hydrochloride salt of this compound. This process includes the use of specific reagents and conditions to ensure the stability and purity of the final product . Another method involves the preparation of dry-granulated pharmaceutical compositions of this compound by compacting this compound hydrochloride with pharmaceutically acceptable excipients . Additionally, a nano-preparation method involves the use of N, N-dimethylformamide and polyvinylpyrrolidone to create a stable nano-drug slurry, which is then spray-dried to obtain the final product .

Nilotinib undergoes various chemical reactions, including oxidation, reduction, and substitution. Common reagents used in these reactions include oxidizing agents, reducing agents, and nucleophiles. For example, this compound can be synthesized via benzanilide formation in water using native chemical ligation (NCL) chemistry . This method involves the coupling of benzoyl and mercaptoaniline fragments to form aromatic amide bonds. The major products formed from these reactions include various derivatives of this compound with potential therapeutic applications.

Treatment of Chronic Myeloid Leukemia

Clinical Efficacy
Nilotinib has been extensively studied for its efficacy in treating newly diagnosed chronic-phase CML. The ENESTnd study demonstrated that patients receiving this compound achieved significantly higher rates of major molecular response compared to those treated with imatinib. Specifically, at 12 months, the rates were 44% and 43% for this compound doses of 300 mg and 400 mg twice daily, respectively, versus 22% for imatinib (p < 0.001) .

Long-Term Outcomes
In a phase 2 study involving 122 patients treated with this compound (400 mg twice daily), long-term follow-up indicated that 91% achieved a complete cytogenetic response, with significant event-free survival rates at 89% and overall survival rates at 93% after five years .

Table 1: Summary of Clinical Trials for this compound in CML

Potential Applications in Neurodegenerative Diseases

Parkinson's Disease
this compound is being investigated for its potential to alleviate symptoms of Parkinson's disease. A phase II trial indicated that while this compound was generally safe and tolerable, it did not demonstrate significant clinical benefits compared to placebo . The study involved multiple sites and assessed safety across different dosages, revealing more adverse events than the placebo group.

Alzheimer's Disease
Research has also explored this compound's neuroprotective properties in Alzheimer's disease. Preclinical studies suggest that this compound can reduce levels of amyloid-β and prevent neuronal degeneration by inhibiting c-Abl signaling pathways . However, clinical trials have yet to yield robust evidence supporting its efficacy in humans.

Table 2: Summary of this compound Studies in Neurodegenerative Diseases

Safety Profile and Adverse Effects

While this compound is effective against CML, its safety profile warrants attention. Common adverse effects include cardiovascular events, rash, elevated bilirubin, and aminotransferases . In studies involving CML patients, treatment discontinuation due to toxicity occurred in approximately 19% of cases.

Table 3: Adverse Effects Reported in Clinical Trials

Nilotinib exerts its effects by selectively inhibiting the BCR-ABL tyrosine kinase, c-KIT, and platelet-derived growth factor receptor (PDGFR) . It binds to the ATP-binding site of BCR-ABL and inhibits its tyrosine kinase activity, thereby preventing the phosphorylation and activation of downstream signaling pathways involved in cell proliferation and survival. This compound is particularly effective in cases of chronic myeloid leukemia that are resistant to imatinib, another tyrosine kinase inhibitor .

Nilotinib is often compared with other tyrosine kinase inhibitors such as imatinib and dasatinib. While imatinib was the first tyrosine kinase inhibitor approved for the treatment of chronic myeloid leukemia, this compound and dasatinib are second-generation inhibitors with improved efficacy and safety profiles . This compound has a higher binding affinity for the BCR-ABL tyrosine kinase and is effective against a broader range of BCR-ABL mutations compared to imatinib . Dasatinib, on the other hand, is structurally distinct from this compound and has a different spectrum of activity against BCR-ABL kinase domain mutants . Other similar compounds include ponatinib, which is effective against the T315I mutation that is resistant to both this compound and dasatinib .

Nilotinib is a potent, second-generation tyrosine kinase inhibitor (TKI) primarily used in the treatment of chronic myeloid leukemia (CML). It selectively inhibits the BCR-ABL1 fusion protein, which is responsible for the uncontrolled proliferation of myeloid cells in CML. This article explores the biological activity of this compound, focusing on its mechanisms of action, effects on various cell types, clinical efficacy, and associated adverse effects.

This compound works by binding to the ATP-binding site of the BCR-ABL1 protein, inhibiting its kinase activity. This inhibition prevents the phosphorylation of downstream signaling molecules involved in cell proliferation and survival pathways. The specificity of this compound for the BCR-ABL1 kinase also contributes to its effectiveness and reduced off-target effects compared to earlier TKIs like imatinib.

Effects on Endothelial Cells

Recent studies have highlighted this compound's impact on endothelial cell function, which is critical for vascular health. Research utilizing human induced pluripotent stem cells (hiPSCs) has shown that this compound adversely affects endothelial cell proliferation and migration while increasing intracellular nitric oxide levels. Notably, these effects were independent of ABL1 inhibition, suggesting that this compound may exert off-target effects that compromise endothelial function and contribute to this compound-induced arterial disease (NAD) in some patients .

Clinical Efficacy

This compound has demonstrated significant efficacy in treating CML. In the ENEST1st trial, which included 1089 patients with newly diagnosed CML in chronic phase, this compound achieved a major molecular response (MMR) rate of 38.4% at 18 months . Long-term follow-up studies have confirmed sustained efficacy, with 91% of patients achieving a complete cytogenetic response and a significant proportion reaching deep molecular responses (MR4 and MR4.5) .

Case Study: Long-Term Efficacy

A phase 2 study involving 122 patients treated with this compound at 400 mg twice daily reported:

• Event-Free Survival : 89% at 5 years
• Overall Survival : 93% at 5 years
• Sustained MR4.5 : Achieved by 59% of patients beyond 2 years
• Adverse Events : Cardiovascular events occurred in 10% of patients .

Adverse Effects

Despite its efficacy, this compound is associated with several adverse effects, particularly cardiovascular complications. In clinical trials, ischemic cardiovascular events were reported in approximately 6-10% of patients . Other common side effects include:

• Rash: 55%
• Elevated bilirubin: 57%
• Elevated aminotransferases: 48%

These adverse effects necessitate careful monitoring during treatment.

Basic Research Questions

Q. What are the primary methodologies for investigating nilotinib’s mechanism of action in preclinical studies?

• Methodological Answer: Preclinical studies should employ kinase inhibition assays (e.g., KINOMEscan® platform) to profile this compound’s selectivity across 442 kinases . Cell culture models (e.g., endothelial angiogenesis assays) and in vivo murine models are critical for validating on-target BCR-ABL inhibition and off-target effects (e.g., EGFR or FRK signaling) . Ensure characterization of pharmacokinetics, including cerebrospinal fluid (CSF) penetration, to assess bioavailability in neurological studies .

Q. How should researchers design phase 3 trials to evaluate this compound’s efficacy in chronic myeloid leukemia (CML)?

• Methodological Answer: Use randomized controlled trials (RCTs) with comparator arms (e.g., imatinib or dasatinib) and predefined endpoints: major cytogenetic response (MCyR), complete cytogenetic response (CCyR), and progression-free survival (PFS). Account for cross-trial variability by standardizing inclusion criteria (e.g., newly diagnosed chronic-phase CML) and adjusting for geographical/ethnic differences in patient responses .

Q. What statistical methods are recommended for analyzing this compound’s safety and tolerability data?

• Methodological Answer: Apply survival analysis (Kaplan-Meier curves) for time-to-event outcomes (e.g., cardiotoxicity incidence). Use multivariate regression to control for confounders like age, prior cardiovascular disease, or genetic polymorphisms linked to KCNH2 channel dysfunction . Report adverse events with Clopper-Pearson confidence intervals for binary outcomes .

Advanced Research Questions

Q. How can conflicting data on this compound’s cardiotoxicity be resolved in meta-analyses?

• Methodological Answer: Conduct systematic reviews with strict inclusion criteria (e.g., 10-year span, peer-reviewed phase 3 trials) . Stratify studies by patient subgroups (e.g., those with preexisting cardiovascular risk factors) and use GRADE criteria to assess evidence quality. Highlight genetic predispositions (e.g., KCNH2 variants) that may explain heterogeneous toxicity profiles .

Q. What experimental strategies differentiate this compound’s efficacy from other BCR-ABL inhibitors in mutation-driven resistance?

• Methodological Answer: Perform in vitro kinase assays to compare this compound’s binding affinity against BCR-ABL mutants (e.g., T315I) with dasatinib or ponatinib . Use structural biology (e.g., X-ray crystallography) to map resistance-conferring mutations. In clinical studies, prioritize patients with confirmed mutation status via Sanger sequencing or ddPCR .

Q. How can researchers address this compound’s off-target effects on angiogenesis in endothelial cell models?

• Methodological Answer: Employ VEGF-induced angiogenesis assays and phosphoproteomics to identify signaling pathways (e.g., VEGFR2 inhibition by ponatinib vs. EGFR activation by this compound) . Validate findings with siRNA knockdown of proposed off-target kinases (e.g., FRK) and correlate with clinical biomarkers (e.g., HVA/DOAPC levels in CSF) .

Q. What are best practices for reconciling cross-trial discrepancies in this compound’s long-term outcomes?

• Methodological Answer: Use individual patient data (IPD) meta-analysis to harmonize endpoints (e.g., PFS definitions) and adjust for trial design heterogeneity (e.g., dosing schedules, follow-up duration) . Apply counterfactual frameworks to estimate outcomes if crossover between trial arms (e.g., imatinib to this compound) had not occurred .

Q. Methodological Frameworks

Q. How to formulate a PICOT research question for this compound’s role in Parkinson’s disease (PD)?

• Population: Patients with PD and CSF-confirmed dopamine deficiency.
• Intervention: this compound 150–300 mg/day.
• Comparison: Placebo or standard PD therapy.
• Outcome: Change in CSF HVA/DOAPC levels at 12 months.
• Time: 24-month follow-up for safety/tolerability .

Q. What guidelines ensure reproducibility in this compound preclinical studies?

• Follow NIH preclinical reporting standards (e.g., ARRIVE guidelines) for animal studies, including valve calcification assays in murine models . For cell-based work, document passage numbers, serum batches, and assay controls (e.g., imatinib as a comparator) .

Q. Data Analysis and Interpretation

Q. How to interpret this compound’s dual role in promoting autophagy and valvular calcification?

• Use transcriptomics (RNA-seq) to identify gene networks (e.g., osteogenic vs. autophagic pathways) in valvular interstitial cells . Validate with immunohistochemistry (e.g., LC3B for autophagy, RUNX2 for calcification) and correlate findings with clinical echocardiography data .