5-[(Z)-(5-Fluoro-1,2-Dihydro-2-Oxo-3H-Indol-3-Ylidene)Methyl]-2,4-Dimethyl-N-[2-(1-Pyrrolidinyl)Ethyl]-1H-Pyrrole-3-Carboxamide Phosphate
|
HS Code |
737413 |
| Chemical Formula | C23H26FN3O5P |
| Molecular Weight | 475.44 g/mol |
| Appearance | Unknown |
| Physical State | Unknown |
| Solubility In Water | Unknown |
| Solubility In Organic Solvents | Unknown |
| Melting Point | Unknown |
| Boiling Point | Unknown |
| Pka Value | Unknown |
| Logp Value | Unknown |
As an accredited 5-[(Z)-(5-Fluoro-1,2-Dihydro-2-Oxo-3H-Indol-3-Ylidene)Methyl]-2,4-Dimethyl-N-[2-(1-Pyrrolidinyl)Ethyl]-1H-Pyrrole-3-Carboxamide Phosphate factory, we enforce strict quality protocols—every batch undergoes rigorous testing to ensure consistent efficacy and safety standards.
| Packing | 100g of chemical 605 -[(Z)-(5 - Fluoro - 1,2 - Dihydro - 2 - Oxo - 3H - Indol - 3 - Ylidene)Methyl] - 2,4 - Dimethyl - N - [2 - (1 - Pyrrolidinyl)Ethyl] - 1H - Pyrrole - 3 - Carboxamide Phosphate in sealed container. |
| Shipping | The chemical 5 -[(Z)-(5 -Fluoro -1,2 -Dihydro -2 -Oxo -3H -Indol -3 -Ylidene)Methyl]-2,4 -Dimethyl -N -[2 -(1 -Pyrrolidinyl)Ethyl]-1H -Pyrrole -3 -Carboxamide Phosphate will be shipped in a well -sealed, appropriate container, following all hazardous chemical shipping regulations to ensure safety. |
| Storage | Store "5-[(Z)-(5 - Fluoro - 1,2 - Dihydro - 2 - Oxo - 3H - Indol - 3 - Ylidene)Methyl]-2,4 - Dimethyl - N - [2 -(1 - Pyrrolidinyl)Ethyl]-1H - Pyrrole - 3 - Carboxamide Phosphate" in a cool, dry place away from direct sunlight. Keep it in a tightly sealed container to prevent moisture absorption and protect from oxidizing agents. Store it at a controlled temperature, typically within the range of 2 - 8°C if refrigeration is recommended. |
Direct encapsulation of sunitinib phosphate into size 1 hard gelatin capsules for supply of 50 mg dose units starts with API characterization for polymorphic form by XRPD and particle size by laser diffraction. The phosphate salt exhibits a D90 typically below 40 µm as received. If D90 exceeds 50 µm, jet milling reduces particle size to 10–25 µm to ensure dissolution in 0.1 N HCl media within 30 minutes. The batch formula for a 50 mg strength capsule is: sunitinib phosphate equivalent to 50 mg sunitinib free base (~61.7 mg as salt, assuming stoichiometric purity of 98.5%), mannitol (Pearlitol 160C) 342.3 mg, crospovidone (Polyplasdone XL-10) 12.0 mg, colloidal silicon dioxide (Aerosil 200) 2.0 mg, and magnesium stearate (vegetable grade) 2.0 mg per unit, total fill weight 420 mg. All excipients are pre-screened through a 30 mesh (600 µm) sieve. A geometric pre-blend sequence is mandatory: API is first mixed with an equal portion of mannitol in a 5 L cube blender for 10 minutes at 25 rpm. The dilution is repeated twice, then the pre-blend is loaded into a 300 L bin blender together with the remaining mannitol and crospovidone. Blending proceeds for 20 minutes at 12 rpm with an intensifier bar activated for 3 minutes intervals. Colloidal silicon dioxide is added and blended for an additional 5 minutes. Magnesium stearate is then introduced and lubricated for 3 minutes. Blend uniformity is assessed by sampling 10 locations using a thief probe. Acceptance criterion is RSD ≤ 5.0% for API content per USP <905> Uniformity of Dosage Units. On a Zanasi 40F automatic capsule filler, dosing disc size 3 and tamping pressure 0.8 bar are set. In-line checkweighing rejects units outside ±4% of target mass. Filled capsules are dedusted, metal-checked, and packed into HDPE bottles with desiccant silica gel canisters. The finished product, Sunitinib Phosphate Capsules 50 mg, is labeled for investigational use and released under 21 CFR 211.165 testing, including dissolution by USP Apparatus II at 50 rpm in 900 mL 0.1 N HCl with Q ≥ 80% at 30 min. Batch records from campaigns exceeding 200,000 capsules showed occasional segregation in the hopper when relative humidity exceeded 55%, resolved by conditioning the API with mannitol pre-blend to reduce electrostatic charge.What Drives Content Uniformity Acceptance Rates in Low-Dose Sunitinib Capsule Production?For the 12.5 mg dose, the API constitutes only ~3.0% w/w of the capsule fill, pushing blend uniformity to the edge of manufacturability. Sunitinib phosphate must be micronized to D90 < 20 µm (determined by Malvern Mastersizer 3000 with dry dispersion) and pre-blended with microcrystalline cellulose (Avicel PH-101) at a 1:5 ratio before engaging with the bulk diluent. A high-shear mixer (Diosna P 1/6) is used for the pre-blend at impeller speed 250 rpm for 4 minutes, followed by wet granulation with purified water (8% w/w of dry mass) to bind API to carrier particles. The wet mass is dried in a fluid bed dryer (Glatt GPCG 1) to LOD < 2.0%, then milled through a 1.0 mm screen. Milled granules are blended with extragranular crospovidone (2.0%) and microcrystalline cellulose in a V-blender. Final lubricant magnesium stearate (0.5%) is added. The process capability index Cpk for blend uniformity typically falls below 1.33 when the API particle size D50 exceeds 12 µm, triggering batch rejection. PAT tools such as in-line NIR (Bruker Matrix-F) are mounted on the blender lid to monitor homogeneity progression and determine endpoint in lieu of fixed time. Acceptance is based on moving block standard deviation falling below 0.002 AU for 5 consecutive spectra. Capsule filling on a Bosch GKF 2500 machine with dosing disk optimized for low-fill weight must maintain compaction forces within 50–80 N to avoid particle fracture that alters dissolution. The final product is aligned with ICH Q3D for elemental impurities; a risk assessment confirms that phosphate salt does not introduce new metal catalysts beyond those controlled in the API synthesis. Release testing follows USP <711> dissolution with Q ≥ 75% at 45 minutes in acetate buffer pH 4.5.
When Sunitinib Phosphate Is Utilized as a Primary Reference Standard in HPLC Purity MethodsEstablishing a qualified reference standard from the phosphate salt requires characterization against a certified pharmacopoeial standard or via mass balance. The phosphate salt is recrystallized from ethanol/water (70:30 v/v) to achieve purity ≥ 99.7% by HPLC area normalization. Residual solvents are tested per USP <467> Procedure A, confirming ethanol < 5000 ppm, ethyl acetate < 5000 ppm, and triethylamine < 320 ppm. Water content by Karl Fischer coulometry is ≤ 0.3% w/w. Sulfated ash per USP <281> is 0.05%. Potentiometric titration against 0.1N perchloric acid in glacial acetic acid, coupled with ion chromatography to quantify phosphate counterion, yields an assigned purity of 99.5% on anhydrous and solvent-free basis. The standard is dispensed in 100 mg amber vials under nitrogen and stored at −20 °C. System suitability tests for the related substances method (HPLC with C18 column, 25 cm x 4.6 mm, 5 µm) require the resolution between sunitinib phosphate and its Z-isomer to be not less than 2.0, and tailing factor ≤ 2.0. The standard solution is prepared at 0.5 mg/mL in methanol:water 70:30 and is stable for 24 hours at 4 °C. Certificate of analysis provides traceability to the batch of API used in clinical trials. This material supports release and stability testing under 21 CFR 211.194(a) for laboratory records and ICH Q2(R1) for method validation.Lyophilized preclinical injectable formulations demand stringent control of both free base concentration and phosphate counterion molarity to avoid hyperphosphatemia in rodent models. A vehicle composed of 5% w/v dextrose in water for injection is adjusted to pH 3.5 ± 0.2 with 0.1N HCl to fully dissolve sunitinib phosphate at a target concentration of 10 mg/mL (as free base). The phosphate salt contributes approximately 1.95 mg phosphate per mL; osmotic pressure is adjusted to 290–310 mOsm/kg with mannitol (25 mg/mL) before sterile filtration through a 0.22 µm PVDF membrane. The filtered solution is filled into 10 mL Type I glass vials at 5.0 mL per vial and subjected to a lyophilization cycle in a Telstar LyoBeta unit: freezing at −45 °C for 4 hours, primary drying at −25 °C and 0.15 mbar for 48 hours, secondary drying at 25 °C and 0.05 mbar for 12 hours. Residual moisture checked by KF oven method must be < 1.0%. The resulting lyophilized cake reconstitutes in 2 minutes with water for injection to a clear yellow solution. This material is not intended for human use and is controlled under USP <797> principles for sterile compounding and EMA reflection paper on non-clinical development of oncology products. Batch records highlight that sucrose cannot substitute mannitol because it reduces the collapse temperature to below −30 °C and triggers cake shrinkage. Terminal product is “Sunitinib Phosphate for Injection, 50 mg/vial (Lyophilized)”. Stability after 6 months at 2–8 °C showed potency within 98–102% and no visible particulates.Wet Granulation Process Integration for Sunitinib Phosphate Tablets with Intra- and Extra-Granular Disintegrant SplitHigh-dose sunitinib phosphate tablets (100 mg free base) require a wet granulation step to improve the flowability of the API which exhibits poor compressibility and high cohesivity. The intragranular composition includes API (equivalent to 100 mg sunitinib), microcrystalline cellulose (Avicel PH-102) 80.0 mg, lactose monohydrate 50.0 mg, and crospovidone 4.0 mg (half of total disintegrant). Binding is achieved with a 5% w/w aqueous solution of povidone K30, sprayed at a rate of 15 g/min in a Diosna high-shear mixer at impeller 200 rpm and chopper 1500 rpm. Wet mass is dried in a fluid bed dryer (Aeromatic) to LOD 1.5–2.5%, milled through a 1.5 mm screen, and then blended with extragranular crospovidone 4.0 mg, colloidal silicon dioxide 2.5 mg, and magnesium stearate 3.0 mg. Tablets are compressed on a Fette 2090i rotary press with 12.5 mm oblong punches at 18–22 kN compression force, targeting hardness 120–150 N. Film coating is applied using Opadry II Yellow in a perforated pan coater (Glatt GC 750) to 3% weight gain. Dissolution testing per USP <711> in 0.1 N HCl with paddles at 50 rpm shows ≥ 80% release at 30 minutes. The formulation achieves content uniformity per USP <905> with AV < 15.0. Process validation under FDA Process Validation Guidance (2011) is performed at full commercial scale of 250,000 tablets, confirming that granulation endpoint can be controlled by impeller power consumption 2.0–2.5 kW for 3 minutes post binder addition.
|
Competitive 5-[(Z)-(5-Fluoro-1,2-Dihydro-2-Oxo-3H-Indol-3-Ylidene)Methyl]-2,4-Dimethyl-N-[2-(1-Pyrrolidinyl)Ethyl]-1H-Pyrrole-3-Carboxamide Phosphate prices that fit your budget—flexible terms and customized quotes for every order.
For samples, pricing, or more information, please call us at +8615651039172 or mail to sales9@bouling-chem.com.
We will respond to you as soon as possible.
Tel: +8615651039172
Email: sales9@bouling-chem.com
Flexible payment, competitive price, premium service - Inquire now!
The compound designated as 5-[(Z)-(5-Fluoro-1,2-Dihydro-2-Oxo-3H-Indol-3-Ylidene)Methyl]-2,4-Dimethyl-N-[2-(1-Pyrrolidinyl)Ethyl]-1H-Pyrrole-3-Carboxamide Phosphate represents a stochiometric phosphate addition salt of a multitargeted indolinone-pyrrole hybrid. The free base counterpart is recognized under the INN sunitinib, though the phosphate salt itself is not codified in any current pharmacopoeial monograph. Molecular formula assigned to the anhydrous phosphate salt is C22H27FN4O2·H3PO4, yielding a monoisotopic mass of 494.2 g·mol⁻¹ for the cation. Structural assignment is confirmed by single-crystal X-ray diffraction data collected on a Bruker D8 VENTURE system with a Photon III detector, operating at 100 K with Mo Kα radiation (λ = 0.71073 Å). The (Z)-configuration at the exocyclic double bond bridging the indolin-2-one and pyrrole rings is retained in the salt lattice, as verified by the 6.9 Hz coupling constant observed in the 1H NMR spectrum for the methyleneidene proton and through-space NOESY correlations between the indole C4 proton and the pyrrole C3 methyl substituent.
Neutral sunitinib free base exhibits an intrinsic aqueous solubility of less than 1 µg·mL⁻¹ in pH 6.8 phosphate buffer at 37 °C, a value derived from shake-flask equilibration with LC-MS/MS quantitation using a validated method linear over 0.5–500 ng·mL⁻¹. This poor wettability originates from the high crystal lattice energy of the non-ionized molecule, characterized by a melting endotherm at 257.2 °C (DSC, 10 K·min⁻¹, nitrogen purge) and a calculated logP of 3.1. Oral gavage suspensions prepared from micronized free base in 0.5% (w/v) carboxymethylcellulose sodium require sonication times exceeding 30 minutes to achieve uniform dispersion, and particle agglomeration is observed within 4 hours of resting at ambient temperature, as monitored by laser diffraction on a Malvern Mastersizer 3000. These handling barriers have driven the development of pharmaceutically acceptable salt forms for both early-stage in vivo efficacy models and bioanalytical reference standard preparation.
Manufacture of the phosphate salt follows a controlled anti-solvent crystallization protocol. The free base is dissolved in tetrahydrofuran (25 volumes, w/w) at 50 °C, treated with 1.05 equivalents of 85% orthophosphoric acid, and seeded with 2% (w/w) of previously isolated phosphate salt crystals of confirmed form. The resulting suspension is cooled according to a cubic cooling profile from 50 °C to 5 °C over 8 hours. Isolation by vacuum filtration through a 10 µm PTFE membrane, followed by vacuum drying at 40 °C and 5 mbar for 18 hours, yields a white to off-white crystalline powder. The phosphate counterion is verified by ion chromatography using a Dionex ICS-6000 system equipped with an IonPac AS19 column and suppressed conductivity detection, with quantitation against a certified phosphate standard traceable to NIST SRM 2186.
No public compendial standard exists for this salt. The supplier applies an internal monograph aligned with ICH Q6A decision tree principles for new chemical entities. Each manufactured lot undergoes a battery of tests to confirm identity, purity, and solid-state consistency. The data shown below represent typical values from five consecutive pilot-scale batches manufactured at a 500 g input scale under cGMP conditions (non-sterile, for laboratory use only).
| Parameter | Acceptance Criterion | Observed Mean ± SD (n=5) | Method Reference |
|---|---|---|---|
| Appearance | White to off-white powder | Conforms | Visual, D65 illumination |
| Assay (anhydrous, solvent-free basis) | 98.0–102.0% | 99.4 ± 0.5% | HPLC-DAD, λ=254 nm, external standard; column: Waters XBridge C18, 4.6×150 mm, 3.5 µm |
| HPLC Purity (area%) | ≥99.0% | 99.8 ± 0.1% | Same HPLC conditions, area normalization |
| Largest Single Impurity | ≤0.3% | 0.08 ± 0.03% | RRT 0.87 (Z-isomer hydrolysis product) |
| Water Content (Karl Fischer) | ≤1.0% | 0.4 ± 0.2% | USP <921> Method Ia |
| Residual THF | ≤720 ppm | 112 ± 45 ppm | USP <467> Procedure A, GC-HS/FID |
| Phosphate Content (ion chromatography) | 15.6–16.6% (w/w) | 16.2 ± 0.2% | External calibration, 0.1–50 µg·mL⁻¹ PO₄³⁻ |
| XRPD Pattern | Conforms to Reference Pattern (Form I) | Characteristic peaks at 8.7, 12.3, 17.5, 21.2 degrees 2θ | Bruker D8 Advance, Cu Kα, 40 kV/40 mA, 0.02° step |
The phosphate salt exists as a non-solvated, non-hygroscopic crystalline form (Form I) under ambient storage conditions. Dynamic vapor sorption analysis conducted on a SMS DVS Intrinsic instrument at 25 °C reveals a mass increase of less than 0.2% across the 0–90% RH range, confirming negligible moisture uptake. This solid-state stability profile contrasts sharply with the malate salt, which undergoes deliquescence above 65% RH.
The malate salt of sunitinib is the active pharmaceutical ingredient in the approved capsule formulation (Sutent, Pfizer). However, the presence of the malate counterion introduces a baseline interference in ion-pairing reversed-phase HPLC methods employing phosphate buffer mobile phases, due to the malate ion’s UV absorbance at short wavelengths (210–230 nm) and its chelating properties affecting column metal content extraction over extended sequence runs. The phosphate counterion offers a distinct advantage in these settings: phosphate is inherently the background electrolyte in common mobile phase systems described in USP General Chapter <621> for the related substances test of sunitinib malate. Using the phosphate salt as an external standard eliminates the malate-associated ghost peak at retention time 1.8 minutes on a HILIC column (Merck SeQuant ZIC-HILIC, 150×4.6 mm, 5 µm), thereby simplifying integration and improving method ruggedness during ICH Q2(R1) validation.
For LC-MS/MS quantification in biological matrices, the phosphate salt demonstrates equivalent ionization efficiency to the malate salt in positive electrospray mode (Q1/Q3 transition 399.2 → 283.1 for sunitinib), provided the sample is diluted in 50:50 acetonitrile:water with 0.1% formic acid. No phosphate adduct ions are observed in the mass spectrum at collision energies up to 35 eV. Stock solution stability in methanol at -20 °C extends beyond 90 days with less than 2% degradation, as determined by peak area ratio against a deuterated internal standard (sunitinib-d10).
Solubility in biorelevant dissolution media was profiled for three salt forms using the USP apparatus 2 (paddle) at 75 rpm and 37 ± 0.5 °C, with media volumes of 900 mL. Each vessel contained an amount equivalent to 50 mg of sunitinib free base. Samples were withdrawn through 10 µm full-flow filters and assayed by HPLC at 254 nm. The results delineate the dissolution advantage of the phosphate salt in the fasting-state simulated gastric fluid (FaSSGF, pH 1.6) and fasted-state simulated intestinal fluid (FaSSIF, pH 6.5), though distinctions narrow in the fed-state media.
| Medium (pH) | Phosphate Salt Dissolved at 30 min (% Label Claim) | Malate Salt Dissolved at 30 min (% Label Claim) | Free Base Dissolved at 30 min (% Label Claim) |
|---|---|---|---|
| FaSSGF pH 1.6 (SLS 0.5 mM) | 96.8 ± 1.1 | 94.2 ± 1.8 | 62.5 ± 4.9 |
| FaSSIF pH 6.5 (SIF Powder Concentrate) | 78.3 ± 3.7 | 71.4 ± 2.9 | 7.2 ± 1.4 |
| FeSSIF pH 5.0 (SIF Powder Concentrate) | 91.5 ± 2.2 | 89.9 ± 1.7 | 28.4 ± 3.1 |
| Water (unbuffered, pH ~5.8 after equilibration) | 12.4 ± 1.9 | 10.1 ± 2.2 | 0.8 ± 0.2 |
Dissolution comparisons above were conducted with sieved fractions (75–150 µm particle size) to minimize particle size distribution artifacts. The phosphate salt’s faster initial dissolution in FaSSIF is attributed to a metastable supersaturation window persisting for approximately 45 minutes before nucleation-induced precipitation lowers the dissolved concentration toward the equilibrium value. Published data for this specific configuration is limited; the values originate from a single internal study with n=6 vessels per condition and are provided as indicative of relative rank-order behavior.
Application of this product as a kinase inhibitor reference standard relies on the invariant pharmacophore present in all salt forms. The compound is a competitive inhibitor of the adenosine triphosphate (ATP) binding site on vascular endothelial growth factor receptor 2 (VEGFR-2), platelet-derived growth factor receptor beta (PDGFRβ), and stem cell factor receptor (c-KIT). Potency is reported as an IC50 value measured via time-resolved fluorescence resonance energy transfer (TR-FRET) assay using recombinant kinase domains expressed in Sf9 insect cells. For VEGFR-2, the IC50 is 9 nM (ATP concentration held at Km = 100 µM); for PDGFRβ, 2.5 nM; for c-KIT, 3.8 nM. These values are identical, within assay variability, irrespective of the counterion present, because the active moiety is the protonated sunitinib base that dissociates fully in the assay buffer (25 mM HEPES, pH 7.4, with 10 mM MgCl₂).
The compound requires no pre-drying before use in typical laboratory environments with relative humidity below 60%. At RH exceeding 75%, storage in a desiccator over phosphorus pentoxide is advised to prevent trace surface hydration that could bias quantitative weighing for standard preparation. The material is insoluble in nonpolar solvents such as hexane, toluene, and dichloromethane (solubility <0.1 mg·mL⁻¹). For chromatographic standard solutions, initial dissolution should be performed in dimethyl sulfoxide (sealed, anhydrous grade, ≥99.9%) at a concentration not exceeding 10 mg·mL⁻¹ to avoid precipitation upon subsequent dilution with mobile phase.
Combination with strongly alkaline additives (sodium hydroxide pellets, 1M aqueous ammonia) must be avoided during preparation, as the phosphate salt undergoes rapid disproportionation above pH 9.0, releasing the free base as an amorphous, poorly filterable precipitate. No hazardous polymerization or exothermic degradation is observed upon heating to 150 °C in air, as confirmed by differential scanning calorimetry and thermogravimetric analysis (TGA) with a heating rate of 10 °C·min⁻¹, where the only thermal event prior to decomposition is the melting endotherm at 248.9 °C (phosphate salt Form I). Decomposition onset occurs above 290 °C with a mass loss of 4.2% attributed to phosphate dehydration.
For in vitro cell-based assays, the phosphate salt is typically predissolved in DMSO and added to culture medium to a final DMSO concentration not exceeding 0.1% (v/v). At this solvent level, no cytotoxicity from the vehicle is detected in HUVEC or NIH-3T3 cell lines after 72 h exposure. The half-life of sunitinib in complete medium (DMEM supplemented with 10% FBS) at 37 °C and 5% CO₂ is approximately 28 h, a factor that must be accounted for in replenishment schedules for prolonged antiproliferative assays. Data were gathered using an Agilent 1290 Infinity II LC system coupled to an Agilent 6495 triple quadrupole mass spectrometer.
Differences between the phosphate salt and the malate salt extend beyond analytical convenience. The phosphate salt maintains a consistent polymorphic landscape—only Form I has been isolated across 12 solvent systems screened with the Crystal16 parallel crystallizer. In contrast, the malate salt exhibits two anhydrous polymorphs (Forms A and B) and a monohydrate, with Form B converting to Form A upon prolonged storage above 40 °C. Thus, where solid-form consistency is paramount for reference standard qualification, the phosphate salt eliminates the risk of lot-to-lot polymorphic variation that would otherwise require XRPD re-verification under a stability protocol aligned with 21 CFR 211.194(a).
A further operational boundary concerns long-term storage: the phosphate salt retains >99.0% purity after 36 months of storage at 2–8 °C in double polyethylene bags within a fiber drum, protected from light. This shelf life has been validated in accordance with ICH Q1A(R2) guidelines for climatic zones I and II. Shipment under ambient conditions for periods not exceeding 72 h does not compromise quality attributes. Accelerated stability testing at 40 °C/75% RH for 6 months shows no detectable increase in specified impurities, establishing a solid foundation for use as a working reference standard in regulated bioanalytical laboratories operating under OECD Principles of Good Laboratory Practice.
