N-(2-Diethylaminoethyl)-5-[(Z)-(5-Fluoro-2-Oxo-Indolin-3-Ylidene)Methyl]-2,4-Dimethyl-1H-Pyrrole-3-Carboxamide
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HS Code |
596314 |
| Chemical Formula | C23H27FN4O3 |
| Molecular Weight | 428.485 g/mol |
| Physical State | Solid (predicted) |
| Boiling Point | 632.9°C at 760 mmHg (predicted) |
| Melting Point | 165 - 167°C |
| Logp | 3.39 (predicted) |
| Pka | 15.05±0.70 (Predicted) |
| Solubility | Soluble in DMSO, sparingly soluble in water |
| Appearance | Yellow - orange powder |
| Stability | Stable under normal conditions, avoid strong oxidizing agents |
As an accredited N-(2-Diethylaminoethyl)-5-[(Z)-(5-Fluoro-2-Oxo-Indolin-3-Ylidene)Methyl]-2,4-Dimethyl-1H-Pyrrole-3-Carboxamide factory, we enforce strict quality protocols—every batch undergoes rigorous testing to ensure consistent efficacy and safety standards.
| Packing | 100g of N-(2 - Diethylaminoethyl)-5-[(Z)-(5 - Fluoro - 2 - Oxo - Indolin - 3 - Ylidene)Methyl]-2,4 - Dimethyl - 1H - Pyrrole - 3 - Carboxamide in sealed container. |
| Shipping | The chemical "N-(2 - Diethylaminoethyl)-5-[(Z)-(5 - Fluoro - 2 - Oxo - Indolin - 3 - Ylidene)Methyl]-2,4 - Dimethyl - 1H - Pyrrole - 3 - Carboxamide" will be shipped in accordance with strict chemical transport regulations, in secure, properly labeled containers to ensure safe transit. |
| Storage | Store “N-(2 - Diethylaminoethyl)-5-[(Z)-(5 - Fluoro - 2 - Oxo - Indolin - 3 - Ylidene)Methyl]-2,4 - Dimethyl - 1H - Pyrrole - 3 - Carboxamide” in a cool, dry place away from direct sunlight. Keep it in a tightly sealed container to prevent moisture absorption and potential reaction with air components. Avoid storing near incompatible substances. |
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Commercial-scale manufacture of sunitinib malate immediate-release capsules begins with micronized active pharmaceutical ingredient having a volume-median particle size D₅₀ ≤ 5 µm and D₉₀ ≤ 20 µm as determined by laser diffraction (ISO 13320:2020). Particle size distribution directly governs dissolution-driven bioavailability; lot-to-lot variability exceeding these limits has been observed to shift tmax by over 45 minutes in fasted-state pharmacokinetic studies. A high-shear wet granulation process is employed: sunitinib malate (equivalent to 12.5 mg, 25 mg, 37.5 mg, or 50 mg of sunitinib free base per unit) is blended with mannitol Ph.Eur./NF as diluent, croscarmellose sodium Ph.Eur./NF as intragranular disintegrant (3.0–5.0% w/w of fill weight), and povidone K30 as binder, then granulated with purified water to achieve a wet mass endpoint with loss-on-drying not exceeding 2.5% after tray drying at 55 °C ± 3 °C. Dried granules are milled through a 0.8 mm oscillating screen, blended with extragranular croscarmellose sodium (1.5–3.0% w/w) and magnesium stearate (1.0–1.5% w/w), and encapsulated into opaque hard gelatin shells using a dosator-type machine with fill weight control of ±3.0% target. Dissolution compliance is verified against an FDA-recommended method: 0.01 N HCl with 1% (w/v) sodium lauryl sulfate, 900 mL, paddle at 75 rpm, with Q = 80% in 30 minutes. Finished product release testing additionally follows USP <905> for content uniformity, with acceptance value AV ≤ 15.0, and related substances by HPLC using an octadecylsilane column (5 µm, 250 × 4.6 mm) with UV 268 nm detection, limiting any single unspecified impurity to ≤ 0.10%. This drug product is registered for the first-line treatment of advanced renal cell carcinoma and imatinib-resistant GIST, with stability assigned under ICH Zone IVb long-term conditions (30 °C/75% RH, 24 months). Compounded Oral Suspensions in Veterinary OncologyExtemporaneously compounded sunitinib oral suspensions are prepared by registered veterinary compounding pharmacies under USP <795> when a commercial human capsule strength does not permit accurate dose titration for canines or felines. A typical vehicle is a 1:1 (v/v) mixture of Ora-Plus® suspending agent and Ora-Sweet® syrup, which provides a near-neutral pH and a palatable viscosity of approximately 350 cP at 25 °C. Sunitinib malate powder is triturated in a glass mortar using geometric dilution to yield a target concentration of 10 mg/mL sunitinib base equivalent; for a 30 mL batch, 400 mg of sunitinib malate salt (containing ~300 mg free base) is weighed on a calibrated balance with ±0.1 mg readability and slowly incorporated into 15 mL of the vehicle paste, then quantitatively transferred to a graduated amber bottle and brought to final volume. Homogeneity must be confirmed by HPLC assay of samples drawn from top, middle, and bottom after 24 hours of equilibration under refrigeration (2–8 °C); acceptance criterion is 90.0–110.0% of labelled strength with RSD ≤ 5.0%. A published stability study indicated 90.3% potency retention after 90 days at controlled room temperature protected from light, so a beyond-use date of 90 days in amber polyethylene terephthalate containers stored at ≤ 25 °C is assigned. This extemporaneous preparation is utilized for the management of canine mast cell tumours with mutant c-KIT, administered orally at doses ranging from 2.0 to 3.5 mg/kg body weight once daily, and is classified as a hazardous drug requiring compliance with USP <800> handling precautions during compounding and dispensing. Why Does Research-Grade Sunitinib Require Strict Solubilisation Protocols?For in vitro kinase inhibition assays, the free base form is dissolved in anhydrous dimethyl sulfoxide (DMSO) to prepare a primary stock solution at 10 mM, a concentration that avoids DMSO cytotoxicity when further diluted to working concentrations of 0.1–5.0 µM in cell culture medium. The powder must be pre-dried under vacuum (40 °C, < 1 mbar) for at least 24 hours because equilibrium moisture content above 0.3% promotes hydrolysis of the amide bond, generating an inactive carboxylic acid metabolite that can confound IC50 determinations. Stock solutions are filtered through a 0.22 µm PVDF syringe filter into sterile amber vials and stored at -20 °C; repeated freeze-thaw cycles must be limited to ≤ 3 to avoid precipitation artefacts. Cell treatment studies targeting phosphorylation of VEGFR2 and PDGFRβ in serum-starved HUVEC monolayers typically expose cells for 1 hour before ligand stimulation, with inhibition quantified via Western blot densitometry normalized to β-actin. All handling is conducted in a BSL-2 biosafety cabinet wearing nitrile gloves tested for breakthrough time ≥ 240 minutes per ASTM D6978-05. The final product is a sterile-filtered, lot-tracked inhibitor solution labelled with preparation date and thiol-reactive impurity warning; it carries an explicit statement ‘Not for human or veterinary therapeutic use’ under 40 CFR Part 720 research exemption. When the Compound Serves as a Certified Pharmacopeial Reference StandardUSP Sunitinib Malate RS (catalogue 1652806) and its European Pharmacopoeia equivalent are issued as certified reference materials for identity, assay, and impurity profiling in finished product quality control. Each lot is characterised by a combination of liquid chromatography with diode-array detection (purity by area normalisation), differential scanning calorimetry (melting endotherm onset at 216 ± 2 °C), thermogravimetric analysis (≤ 0.5% weight loss up to 150 °C), and Karl Fischer coulometric titration for water content (≤ 0.5%). The assigned chromatographic purity value, typically 99.8% ± 0.2% (on the anhydrous, solvent-free basis), is traceable to the SI through a calibrated balance and certified reference materials for the water and residual solvent determinations per ISO 17034:2016. The reference standard is dispensed in amber borosilicate glass vials under argon overlay, each containing 200 mg of micronized solid. For use, 25.0 mg is accurately weighed and dissolved in 50.0 mL of a diluent composed of acetonitrile:water:trifluoroacetic acid (40:60:0.1 v/v/v), yielding a 0.5 mg/mL stock solution; this must be used within 8 hours and protected from light due to Z→E photoisomerisation. Analytical methods specified in USP Monograph “Sunitinib Malate Capsules” employ an L1 column (4.6 mm × 25 cm, 5 µm), flow rate 1.0 mL/min, and detection at 268 nm, with system suitability requiring resolution between sunitinib and its N-oxide impurity of at least 2.0. Liposomal Encapsulation Improves Tumour-Selective DeliveryPreclinical nanoformulation efforts use sunitinib free base as a hydrophobic payload for poly(ethylene glycol)-block-poly(lactic-co-glycolic acid) (PEG-PLGA) nanoprecipitation, aiming to prolong circulation half-life and reduce off-target kinase inhibition in cardiac tissue. The organic phase is prepared by dissolving sunitinib and PEG5k-PLGA45k (lactide:glycolide 85:15) at a drug-to-polymer ratio of 1.5:10 (w/w) in acetone (HPLC-grade, water content ≤ 0.02%). This phase is added dropwise at 0.5 mL/min via syringe pump into 10 volumes of ultrapure water containing 0.25% w/v poly(vinyl alcohol) (87–90% hydrolysed, MW 30–70 kDa) under magnetic stirring at 800 rpm and 25 °C. Acetone is removed under reduced pressure (40 kPa, 35 °C) until its headspace GC level falls below 200 ppm, in compliance with ICH Q3C guideline for Class 3 residual solvents. The resulting nanoparticle dispersion is centrifuged at 12,000 × g for 30 minutes, washed twice, and freeze-dried with 5% (w/w) trehalose as cryoprotectant. Product release criteria include mean hydrodynamic diameter Z-average = 140–180 nm with polydispersity index (PDI) ≤ 0.20 by dynamic light scattering at 173° backscatter angle (ISO 22412:2017), surface charge (zeta potential) between -25 mV and -35 mV in 1 mM NaCl, and encapsulation efficiency ≥ 85% measured after Sephadex G-25 size-exclusion chromatography. Lyophilised cake moisture content is controlled to ≤ 1.2% (coulometric KF), and reconstitution in 0.9% NaCl yields an isotonic injectable dispersion suitable for intravenous administration in murine xenograft models. This batch-scale configuration, operated in a laminar flow isolator meeting ISO Class 5 air cleanliness, is documented for IND-enabling toxicology and proof-of-concept efficacy studies according to 21 CFR 58 (Good Laboratory Practice for nonclinical studies). |
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N-(2-Diethylaminoethyl)-5-[(Z)-(5-fluoro-2-oxoindolin-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-carboxamide, a pyrrole-indolinone congener, functions as a multi-target receptor tyrosine kinase inhibitor with a molecular weight of 398.46 g/mol and CAS registry number 557795-19-4. The (Z)-geometric isomer is the thermodynamically preferred configuration, confirmed by single-crystal X-ray diffraction, and constitutes the sole bioactive species incorporated into pharmaceutical preparations. Its therapeutic mechanism proceeds through competitive ATP-binding pocket occupancy in the split kinase domains of VEGFR-2 (Flk-1/KDR), PDGFR-β, c-KIT, FLT3, and RET, yielding sub-micromolar inhibition constants that translate to anti-angiogenic and direct anti-tumor activities. The free base exhibits practically negligible aqueous solubility across the physiological pH range—measured at ≤ 0.1 µg/mL in water at 25 °C—necessitating salt formation, typically with L-malic acid, to achieve systemic exposure following oral administration. The following sections dissect the pharmacochemical fingerprint of the compound, its differentiation from structurally divergent kinase inhibitors, and the quality attributes governing large-scale manufacture of the active pharmaceutical ingredient.
The indolin-2-one scaffold of the compound contrasts sharply with the benzamide-phenylaminopyrimidine architecture of imatinib and the biaryl urea motif of sorafenib. Kinase profiling against a panel of 16 recombinant human tyrosine kinases using a FRET-based Z'-LYTE® assay platform reveals a multi-target engagement profile that underlies both therapeutic breadth and characteristic adverse events. The compound demonstrates potent inhibition of VEGFR-2 (Kdr) with an IC50 of 80 nM, PDGFR-β at 2 nM, and c-KIT at 1–10 nM, while showing negligible activity against the non-receptor tyrosine kinase ABL (IC50 > 10 µM). In contrast, imatinib potently suppresses BCR-ABL (IC50 25 nM) and wild-type KIT (100 nM), yet its VEGFR-2 inhibition is several orders of magnitude weaker. Sorafenib, carrying a diaryl urea, inhibits VEGFR-2 (90 nM) and also hits C-RAF (6 nM) and B-RAF, an activity absent from the indolinone compound. Differences in off-target activity against AMPK and other metabolic kinases have been linked to the propensity for hypertension and hypothyroidism observed clinically with the pyrrole-carboxamide derivative, toxicity signatures not derived from BCR-ABL suppression. The table below summarizes comparative biochemical IC50 values extracted from publicly available pharmacological datasets.
| Kinase Target | Indolinone Compound (IC50, nM) | Imatinib (IC50, nM) | Sorafenib (IC50, nM) |
|---|---|---|---|
| VEGFR-2 (KDR) | 80 | >10,000 | 90 |
| PDGFR-β | 2 | 100 | 57 |
| c-KIT (wild-type) | 1–10 | 100 | 68 |
| FLT3 | 250 | >5,000 | 58 |
| RET | 100 | — | — |
| B-RAF | >10,000 | — | 22 |
The free base has been reported to crystallize in at least three anhydrous polymorphic modifications and one methanol solvate, though commercial development has converged on the 1:1 L-malate salt, which crystallizes as a thermodynamically stable Form I. Differential scanning calorimetry of the malate salt exhibits a single endothermic melting event with an onset temperature near 205 °C and an enthalpy of fusion corresponding to high crystallinity. Polymorphic interconversion risk is assessed through suspension equilibration in isopropanol/water (95:5 v/v) at 40 °C for 48 hours, with subsequent XRPD analysis against a reference pattern indexed to the triclinic P1 space group. Accelerated stability testing per ICH Q1A(R2) at 40 °C/75% RH in open petri dishes demonstrates no form conversion over 6 months, though moisture uptake at 25 °C/60% RH remains below 0.5% w/w as determined by dynamic vapor sorption. Photodegradation, however, proceeds via isatin ring opening under exposure to light sources conforming to ICH Q1B Option 2, mandating storage in amber glass containers with a light transmission limit of ≤ 10% at 450 nm.
In a validated convergent synthetic route, 5-fluoro-2-oxoindoline-3-carboxaldehyde is condensed with 5-formyl-2,4-dimethyl-1H-pyrrole-3-carboxylic acid (2-diethylaminoethyl)amide in the presence of catalytic piperidine in methanol under reflux at 64–66 °C for 8 hours. The Knoevenagel condensation proceeds with high Z-selectivity, driven by intramolecular hydrogen bonding between the amide N–H and the oxindole carbonyl; the E-isomer, quantified as a specified impurity by chiral HPLC on an amylose tris(3,5-dimethylphenylcarbamate) stationary phase, is controlled at a limit of ≤ 0.15% area. Reaction work-up involves neutralization, aqueous extraction, and crystallization from ethyl acetate/methanol, yielding the free base with a typical purity of ≥ 99.5% by HPLC at 263 nm. Salt formation with L-malic acid in acetone/water (90:10 v/v) at 50 °C generates the malate salt, which is isolated on a centrifuge and dried in a conical vacuum dryer at 45–50 °C under a vacuum of ≤ 10 mbar for 12–18 hours. Process-related impurities include the des-ethyl analog, arising from incomplete alkylation of the diethylaminoethyl side chain, and the isatin oxidation product 5-fluoro-2,3-dioxoindoline, controlled via IPC limits. Genotoxic impurity risk is evaluated per ICH M7(R1) using purge factor calculations; any residual allylic halide intermediates are below the threshold of toxicological concern (1.5 µg/day) based on staged spike-and-purge studies performed at 2000 L glass-lined reactor scale with jacket temperature control at ±2 °C.
Residual solvents are determined by headspace GC-FID using a DB-624 column (30 m × 0.53 mm, 3.0 µm film) with a limit of quantitation validated at 50% of the ICH Q3C(R8) Option 1 concentration limits. Elemental impurities are tested via microwave-assisted acid digestion followed by ICP-MS, with method detection limits verified against the 30% J-value criterion of ICH Q3D Guideline for Elemental Impurities. The compliance matrix for the drug substance is provided below.
| Analyte | Limit (ppm) | Governing Guideline | Analytical Technique |
|---|---|---|---|
| Methanol | ≤ 3000 | ICH Q3C, Class 2 | HS-GC-FID |
| Piperidine | ≤ 720 | ICH Q3C, Class 2 | HS-GC-FID |
| N,N-Dimethylformamide | ≤ 880 | ICH Q3C, Class 2 | HS-GC-FID |
| Ethyl Acetate | ≤ 5000 | ICH Q3C, Class 3 | HS-GC-FID |
| Acetone | ≤ 5000 | ICH Q3C, Class 3 | HS-GC-FID |
| Palladium | ≤ 10 | ICH Q3D, Oral PDE | ICP-MS |
| Nickel | ≤ 20 | ICH Q3D, Oral PDE | ICP-MS |
| Arsenic | ≤ 1.5 | ICH Q3D, Oral PDE | ICP-MS |
The formulated drug product comprises the L-malate salt equivalent to 50 mg of the free base per hard gelatin capsule, diluted with mannitol (Pearlitol® 200 SD), croscarmellose sodium (Ac-Di-Sol®), and magnesium stearate (Ligamed® MF-2-V). Blend uniformity, assessed as per the FDA Guidance for Industry “Powder Blends and Finished Dosage Units—Stratified In-Process Dosage Unit Sampling”, must yield an acceptance value (AV) of ≤ 15.0 across 10 sampling locations within the V-blender. The particle size distribution of the milled drug substance, measured by laser diffraction (Malvern Mastersizer 3000 with Aero S dry dispersion at 1.0 bar), is specified with a D90 of ≤ 30 µm to prevent segregation and ensure rapid dissolution. Dissolution testing is conducted according to USP 711, Apparatus 2 (paddles) at 75 rpm, in 900 mL of 0.1 N HCl containing 0.5% (w/v) sodium lauryl sulfate at 37 ± 0.5 °C. The dissolution criterion (Q) is set at ≥ 80% (Q=80) at 30 minutes. The malate salt’s low glass transition temperature (Tg) and the high drug load (~60% w/w) necessitate pre-screening of capsule shell material for cross-linking potential using forced-degradation samples stored at 40 °C/75% RH in HDPE bottles with desiccant.
Wet granulation of the malate salt formulation on a Fielder PMA 300 high-shear granulator with a 25 L bowl exhibits pronounced sensitivity to water quantity and impeller speed. At an impeller setting of 250 rpm and chopper at 1500 rpm, purified water is added at a rate of 200 g/min through a pressure pot to a total of 28–32% w/w of the dry blend mass. Granulation endpoint is determined by monitoring net power consumption on the main drive motor; a power plateau corresponding to a torque increase of 40–60 N·m above baseline signals the transition from pendular to funicular liquid bridging without over-wetting. The wet mass is immediately passed through a 4.0 mm Quadro Comil and dried in a Glatt GPCG 5 fluid bed dryer with an inlet air temperature of 60 °C, targeting a loss on drying (LOD) of 1.5–2.5% w/w by halogen moisture analyzer at 85 °C. Sticking to tablet punch faces during compression on a 19-station rotary press (Korsch XL 200) at 15–20 kN compression force and a turret speed of 30 rpm is mitigated by external lubrication with magnesium stearate at 0.8–1.2% w/w and the addition of talc at 2.0% w/w as a glidant/anti-adherent. Ejection forces above 600 N are indicative of insufficient lubrication or granule over-drying and trigger process interventions based on real-time compression data analytics recorded via the press’s Kramer electronics module.
