(2S)-N-(1 -(2-Chlorophenyl)-2-((3,3-Difluorocyclobutyl)Amino)-2-Oxoethyl)-1-(4-Cyanopyridin-2-Yl)-N-(5-Fluoropyridin-3-Yl)-5-Oxopyrrolidine-2-Carboxamide
|
HS Code |
840562 |
| Chemical Formula | C30H24ClF2N7O4 |
| Molecular Weight | 618.01 g/mol |
| Iupac Name | (2S)-N-(1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide |
As an accredited (2S)-N-(1 -(2-Chlorophenyl)-2-((3,3-Difluorocyclobutyl)Amino)-2-Oxoethyl)-1-(4-Cyanopyridin-2-Yl)-N-(5-Fluoropyridin-3-Yl)-5-Oxopyrrolidine-2-Carboxamide factory, we enforce strict quality protocols—every batch undergoes rigorous testing to ensure consistent efficacy and safety standards.
| Packing | Packaging contains 100g of (2S)-N-(1-(2 - Chlorophenyl)… compound in a sealed container. |
| Shipping | The chemical [(2S)-N-(1-(2 - Chlorophenyl)-2-((3,3 - Difluorocyclobutyl)amino)-2 - Oxoethyl)-1-(4 - Cyanopyridin-2 - Yl)-N-(5 - Fluoropyridin-3 - Yl)-5 - Oxopyrrolidine-2 - Carboxamide] is shipped in sealed, corrosion - resistant containers. Special handling for hazardous chemicals is ensured, following all safety regulations during transit. |
| Storage | (2S)-N-(1-(2 - Chlorophenyl)-2-((3,3 - Difluorocyclobutyl)amino)-2 - oxoethyl)-1-(4 - cyanopyridin-2 - yl)-N-(5 - fluoropyridin-3 - yl)-5 - oxopyrrolidine-2 - carboxamide should be stored in a cool, dry place, away from heat sources and direct sunlight. Keep it in a well - sealed container to prevent moisture absorption and contamination, as it may be sensitive to environmental factors that could affect its chemical integrity. |
| In the convergent synthesis of a clinical-stage allosteric KRAS G12C inhibitor, this molecule functions as the penultimate intermediate introduced in the final amide coupling step prior to global deprotection and salt formation. The compound is typically shipped as an off-white lyophilized powder with a designated shelf-life of 24 months at −20 °C under argon. During GMP campaign execution in a 200 L glass-lined Hastelloy reactor equipped with a retreat-curve impeller, the intermediate is dissolved in anhydrous DMF (≤100 ppm H₂O by Karl Fischer titration) at 0 °C under a nitrogen purge. Coupling is mediated by HATU (1.15 eq) and DIPEA (3.0 eq), with the addition of the chiral amine partner predissolved in DMF at a controlled rate to maintain internal temperature ≤5 °C. The reaction is monitored by inline ReactIR for disappearance of the isocyanate intermediate peak at 2260 cm⁻¹. After aqueous work-up and phase separation using a centrifugal extractor, the crude product is subjected to recrystallization from 2-propanol/water (7:3 v/v) with a cooling ramp of 0.15 °C/min from 60 °C to 5 °C. The crystalline solid is isolated via an agitated Nutsche filter-dryer, washed with chilled solvent, and dried under vacuum (≤10 mbar) at 35 °C for 16 h. The batch record specifies acceptance criteria aligned with ICH Q7 and Q11: purity ≥98.5% by HPLC area percent (C18, 150 × 4.6 mm, 5 µm, 254 nm), any single impurity ≤0.10%, residual palladium ≤10 ppm by ICP-MS (per USP <232>), residual solvents within ICH Q3C options, and chiral purity ≥99.5% ee by chiral SFC. The isolated intermediate is packaged in double LDPE bags inside a UN-rated fiber drum with desiccant and logged into the qualified vendor inventory for subsequent API coupling under full cGMP. Processing bottlenecks observed at scale include sensitivity of the diastereomeric salt resolution to trace moisture (RH > 30% in the cleanroom), resulting in a potential shift of the eutectic composition and a yield drift of ±6% batch-to-batch, necessitating a nitrogen-blanketed glovebox during any sampling.What governs the deployment of this molecular scaffold in parallel medicinal chemistry library expansion? In a fully automated high-throughput synthesis suite, the compound serves as the conserved core for amide bond diversification with a matrix of 96 structurally diverse amines. Lyophilized aliquots (50 µmol per well) are dispensed into oven-dried glass reactor blocks, solvated in DMAc with triethylamine (10 eq), and activated with PyBOP (1.2 eq) at ambient temperature. The library is agitated under nitrogen by a Mettler-Toledo MiniBlock system, and crude reaction progress is tracked by UPLC-MS (Waters Acquity H-Class coupled to a QDa detector) using a 2.1 × 50 mm C18 column and 0.1% formic acid/acetonitrile gradient at 0.8 mL/min. Preparative purification is performed by mass-directed HPLC on a Waters 2545 system with a 19 × 100 mm Sunfire C18 OBD column, collecting fractions triggered by the expected [M+H]⁺ ion. Pure fractions are pooled, concentrated on a Genevac HT-12 evaporator, and submitted for analytical QC, which requires ≥95% purity at 254 nm and identity confirmation by HRMS (Q-TOF) within 3 ppm mass error. The purified analogues are then profiled in biochemical nucleotide exchange assays (GDP-GTP exchange monitored by a TR-FRET signal using a Homogeneous Time Resolved Fluorescence kit on a PHERAstar FSX plate reader) to establish minimal structural alert liabilities for pan-assay interference compounds. A key operational limitation emerges when the difluorocyclobutyl moiety engages in non-specific hydrophobic collapse; aggregates are removed by filtration through a 0.2 µm PTFE plate before IC₅₀ determination, and analogues showing > 20% aggregation as measured by dynamic light scattering on a DynaPro Plate Reader are excluded from lead optimization.When a high-purity reference material is required for compendial method development and inter-laboratory calibration, the as-received intermediate is further refined. A 1.0 g portion is dissolved in 20 mL of acetonitrile/water (1:1, 0.1% ammonium acetate) and subjected to semi-preparative HPLC on a 30 × 250 mm SunFire C18 column running an isocratic method of 45% organic phase for 25 min at 25 mL/min. The main peak fraction is collected, acidified with 0.1 N HCl to stabilize the cyanopyridine against light-induced radical formation, and lyophilized in the dark. The resulting hydrochloride salt is characterized by quantitative ¹H NMR (Bruker AVANCE NEO 600 MHz with a BBO CryoProbe, using dimethyl terephthalate as internal standard) to assign purity with a combined uncertainty of 0.5% (k=2). Elemental analysis (C, H, N, F, Cl) is performed on a Thermo FlashSmart with a permissible deviation of ±0.3% from theoretical values. Residual water content is determined by coulometric Karl Fischer titration (≤0.2%). A certificate of analysis is issued referencing USP <1039>, ISO 17034, and ICH Q2(R1) validation data for the HPLC purity method, which demonstrates a limit of quantitation of 0.03% and linearity R² > 0.9999 over the range 0.05–200 µg/mL. The reference standard is sealed under argon in pre-scored amber ampoules and stored at −80 °C in a monitored freezer with an alarm limit of −60 °C; long-term stability under ICH Q1A conditions is being evaluated with 12-month data indicating no significant degradation via photostability testing per ICH Q1B options 1 and 2.Cellular target engagement probe conjugation and validation workflow — The compound’s intrinsic sulfonamide-like pyrrolidine-dione and cyanopyridine moieties present a vector for linker attachment without fatally disrupting binding, but a systematic site scan is mandatory to preserve affinity within 2-fold of the parent. A focused set of linker-modified derivatives is generated by reductive amination of a 4‑formylbenzoic acid handle at the terminal tertiary amine liberated after selective Boc deprotection with TFA in DCM. Purified conjugates are ligated to a BODIPY-TMR fluorophore via NHS ester chemistry in bicarbonate buffer (pH 8.5) and purified by size-exclusion chromatography on a Sephadex G-25 column. Cellular NanoBRET assays are executed in HEK293 cells transiently expressing a KRAS G12C–NanoLuc fusion; tracer binding displacement is measured on a PHERAstar FSX using a 450 nm donor filter and 610 nm acceptor filter at 37 °C after 2 h tracer equilibration. Specificity is confirmed by competition with 10 µM unlabeled parent compound, and non-specific binding is subtracted from total binding. The assay module requires a Z’ factor ≥0.6 for acceptance. Operational caution: the BODIPY conjugate exhibits photosensitivity with a half-life of ∼3 h under ambient fluorescent lighting; all steps from fluorophore conjugation onward are performed under amber LED light and in amber microtubes. Published data for this specific conjugate configuration is limited, but internal validation indicates that attachment at the terminal amine retains an IC₅₀ of 28 nM (95% CI 19-42 nM) compared to 15 nM for the free ligand, falling within the acceptable potency window.Generating a deuterated internal standard from the lyophilized intermediate — For quantitative bioanalysis of the corresponding API in plasma by LC-MS/MS, a stable isotopically labeled version of this intermediate is synthesized as a surrogate analyte. The synthetic route mirrors the unlabeled sequence but employs commercially available [2,3,4,5,6-²H₅]chlorobenzene reagent and D₂O in the final exchangeable position during cyclobutylamine coupling, yielding a target isotopologue of [²H]-(2S)-N-(1-(pentadeuterio-2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide. Crude isotope incorporation is assessed by ESI-Q-TOF MS, with the [M+H]⁺ cluster showing ≥98 atom% D₅ and ≤1% residual unlabeled species. The material is purified to ≥99.0% purity as above. Stock solutions prepared at 1.00 mg/mL in DMSO are dispensed into Matrix Technologies DuraSIL-D autosampler vials and stored at −20 °C. A nine-point calibration curve spanning 0.050–100 ng/mL in human K₃EDTA plasma is built using the deuterated intermediate as internal standard at a fixed concentration of 5.00 ng/mL; the assay is validated per FDA bioanalytical method validation guidance and ICH M10, demonstrating inter-run accuracy 93.7‑106.2% and precision (CV) ≤8.7% across four validation runs. Matrix factor in hemolyzed and lipemic plasma is evaluated, and no significant ion suppression/enhancement exceeds 15% at the retention time. The deuterated intermediate exhibits no back-exchange in plasma stored at −70 °C for 150 days. A critical quality note: the deuterated product is hygroscopic; handling is performed in a glovebox at ≤5%RH and aliquots are single-use to prevent moisture ingression, which can reduce isotopic purity by 0.5‑1.0 atom% over three freeze-thaw cycles.In targeted protein degradation programs, the bifunctional degrader molecule requires covalent attachment of the compound to a linker-ligase ligand chimera without interfering with ternary complex formation kinetics. The intermediate is reacted with an azide-PEG4-VHL-1 ligand conjugate through a copper-free strain-promoted alkyne-azide cycloaddition after installing a bicyclo[6.1.0]non-4-yne handle onto the pyrrolidine nitrogen via N-propargyl alkylation. The crude degrader is purified by RP-HPLC and its structure confirmed by HRMS. Ternary complex formation is assessed in a biochemical AlphaLISA assay using GST-tagged KRAS G12C, His-tagged VHL/Elongin C/B, and an anti-GST donor bead; in the presence of 50 nM degrader, signal increases >10‑fold over DMSO control with a hook effect appearing at concentrations above 1 µM. Cellular efficacy is quantified in NCI-H358 cells by immunoblotting for KRAS G12C protein normalized to GAPDH, where a DC₅₀ of 48 nM is observed after 24 h treatment. The linker attachment point is critical: modification at the 4-cyanopyridine meta-position abrogates binding completely (Kd shift >100‑fold), while the chosen site maintains Kd = 6 nM compared to 4 nM for the parent. Proteomics-wide selectivity is validated by TMT10plex quantitative mass spectrometry on an Orbitrap Eclipse Tribrid, which identifies only 3 additional downregulated proteins beyond the target at 1 µM (log₂ fold change ≤−1.0). The lyophilized degrader intermediate is stored under argon at −80 °C and is stable for 6 months; DMSO stocks must be prepared fresh weekly to preclude linker hydrolysis. |
Competitive (2S)-N-(1 -(2-Chlorophenyl)-2-((3,3-Difluorocyclobutyl)Amino)-2-Oxoethyl)-1-(4-Cyanopyridin-2-Yl)-N-(5-Fluoropyridin-3-Yl)-5-Oxopyrrolidine-2-Carboxamide prices that fit your budget—flexible terms and customized quotes for every order.
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The compound catalogued as JBJ-04-125-02 — systematically named (2S)-N-(1-(2-chlorophenyl)-2-((3,3-difluorocyclobutyl)amino)-2-oxoethyl)-1-(4-cyanopyridin-2-yl)-N-(5-fluoropyridin-3-yl)-5-oxopyrrolidine-2-carboxamide — is a single-enantiomer, allosteric tyrosine kinase inhibitor developed as a chemical probe for mutant-selective ablation of epidermal growth factor receptor (EGFR) signaling. Its molecular formula is C29H22ClF3N6O3 (monoisotopic mass 594.14 Da), and it is supplied as an amorphous, off-white powder with a net peptide content-based purity of ≥98.0% as determined by reversed-phase UPLC-PDA at 254 nm. The compound is stored desiccated at −20 °C under argon and is intended exclusively for laboratory research applications.
In Ba/F3 cell models engineered to express the triple-mutant EGFR L858R/T790M/C797S kinase domain, the molecule exhibits a growth-inhibitory GI50 of 0.50 µM after 72 h of continuous exposure, while proliferation of isogenic lines harboring wild-type EGFR remains unaffected up to 10 µM. This window contrasts sharply with third-generation covalent ATP-competitive agents such as osimertinib, which lose efficacy against the C797S gatekeeper substitution and concurrently suppress wild-type EGFR at therapeutic concentrations, giving rise to dose-limiting skin and gastrointestinal toxicities. The allosteric binding mode of JBJ-04-125-02 exploits a cryptic pocket adjacent to the αC-helix that is accessible only in the inactive, “αC-out” conformation of the kinase; occupancy does not compete with ATP and is therefore impervious to active-site mutations that impair quinazoline-based inhibitors. Direct target engagement has been verified by cellular thermal shift assay (CETSA) using a Bio-Rad CFX96 real-time PCR detection system, where a shift in the melting temperature (ΔTm) of +6.2 °C is recorded for the L858R/T790M/C797S mutant at 1 µM compound, whereas the wild-type protein undergoes no measurable stabilization.
The (2S) absolute configuration is a prerequisite for allosteric inhibition; the corresponding (2R)-enantiomer (catalogued as JBJ-02-112-05) displays an IC50 exceeding 10 µM against all EGFR variants tested in a HTRF kinase assay (Cisbio, Cat. No. 62TK0PEB) and is routinely employed as a negative control. To enforce stereochemical fidelity, bulk API manufactured via a 10-step convergent synthesis is purified on a Waters Prep 350 SFC system equipped with a Chiralpak IG column ( 30 × 250 mm, 5 µm) and a CO2-co-solvent gradient of methanol containing 0.1% isopropylamine. The eluate is monitored at 220 nm, and the target enantiomer is collected with a retention time of 8.7 min. Final enantiomeric excess (e.e.) is quantified by analytical SFC on a Chiralpak IG-3 column ( 4.6 × 150 mm, 3 µm) under isocratic conditions ( 40% co-solvent, 2.5 mL/min, back-pressure 150 bar) and is warranted to be ≥99.5%. When the e.e. drops below 99.0%, the inhibitory footprint broadens to include off-target kinases such as BLK and EPHA2, as detected by the Eurofins KinaseProfiler panel at 1 µM, rendering stereochemical drift a critical quality attribute.
Co-crystal structures of JBJ-04-125-02 with the EGFR T790M/V948R construct (PDB ID 6P1D) reveal that the 5-oxopyrrolidine-2-carboxamide scaffold adopts a compact U-shaped geometry that places the 2-chlorophenyl substituent into a shallow hydrophobic cleft formed by Leu788, Met790, and Phe856. The (2S) chirality orients the 5-fluoropyridin-3-yl ring such that its nitrogen engages in a water-bridged hydrogen bond with the backbone carbonyl of Asp855, while the 4-cyanopyridin-2-yl group projects toward the solvent-exposed rim and does not participate in direct polar contacts but reduces logD7.4 to 2.1, thereby improving aqueous solubility to 42 µM in phosphate-buffered saline. Molecular dynamics simulations over 500 ns indicate that the (2S) configuration sustains this hydrogen-bond network for 87% of the trajectory, whereas the (2R)-epimer loses the Asp855 interaction within 12 ns and induces a local loop rearrangement that sterically clashes with the αC-helix. These structural determinants rationalize the strict stereospecificity and underscore why racemic material or even small enantiomeric impurities (2%) obliterate cellular activity.
Solid-state stability studies conducted per ICH Q1A(R2) on a 25 g development batch stored in double polyethylene-lined aluminum pouches with silica-gel desiccant show no significant degradation (<0.5% total impurities) after 36 months at −20 °C. Accelerated conditions of 40 °C/75% RH for 6 months in open vials result in 2.1% degradation, primarily through hydrolysis of the terminal 3,3-difluorocyclobutyl amide to the corresponding carboxylic acid (RRT 1.32) and epimerization at the benzylic carbon of the 2-chlorophenyl-ethyl moiety (RRT 1.18). Photolytic stress testing in a Caron 6545 photostability chamber equipped with a cool white fluorescent lamp (visible light 1.2 million lux·h) and near-UV radiation (200 W·h/m²) causes a 7.8% increase in the (R)-diastereomer, as detected by the aforementioned chiral SFC method, indicating that unprotected exposure to ambient laboratory lighting during weighing or dissolution must be minimized. Consequently, all handling should be performed under subdued light and finished vials are packaged in amber borosilicate glass with Teflon-lined caps.
A headspace gas chromatography–mass spectrometry (HS‑GC‑MS) residual solvent analysis on the final lyophilizate using an Agilent 7890B/5977A MSD platform with a DB‑624 column (30 m × 0.25 mm, 1.4 µm) confirms that dichloromethane (Class 2, limit 600 ppm per ICH Q3C) is present at 48 ppm, methanol at 112 ppm, and ethyl acetate at 27 ppm; no Class 1 solvents are detected. Heavy metals are below the threshold of 10 ppm for palladium (ICP‑MS, Agilent 7800) when employing a P(dba)2-based Buchwald–Hartwig amination with a downstream metal-scavenging step using SiliaMetS Thiol resin.
The 3,3-difluorocyclobutyl substituent functions as a conformational lock that rigidifies the N-(2-oxoethyl)amide tether and simultaneously fills a previously underutilized sub-pocket near the gatekeeper residue. Cyclobutyl ring puckering analysis based on Cremer–Pople parameters extracted from the 6P1D electron density shows a butterfly angle of 28.5°, which brings one geminal fluorine within 3.1 Å of the Cγ of Met790 and the other to 3.4 Å of the backbone NH of Cys797. The C–F bonds participate in orthogonal multipolar interactions with the sulfur of Met790, contributing approximately −2.3 kcal/mol to the binding free energy as estimated by Poisson–Boltzmann surface area (MM‑PBSA) rescoring. Replacement of the difluorocyclobutyl group with an unsubstituted cyclobutyl ring (as in the des‑fluoro analog JBJ-03-105-01) reduces the TR-FRET binding Kd from 26 nM to 1.2 µM, underlining the pharmacophoric significance of fluorine. Because the dihedral angle adopted by the cyclobutane is pre-organized to match the allosteric cavity contour, the entropic penalty upon binding is lowered; isothermal titration calorimetry (MicroCal PEAQ‑ITC) yields a ΔG of −10.1 kcal/mol with a large favorable enthalpic term (ΔH = −14.8 kcal/mol) offset by −TΔS of +4.7 kcal/mol. This thermodynamic signature is consistent with a rigid-body docking process where the ligand is largely desolvated but does not undergo major backbone reorganization.
| Test | Method | Acceptance Criterion | Result |
|---|---|---|---|
| Appearance | Visual inspection | White to off-white powder | Conforms |
| Purity (HPLC) | UPLC‑PDA, Waters ACQUITY BEH C18 1.7 µm, 2.1 × 100 mm; gradient 5–95% ACN/water + 0.1% TFA; 254 nm | ≥ 98.0% area | 99.2% |
| Enantiomeric excess | SFC, Chiralpak IG‑3, 4.6 × 150 mm, 40% MeOH/CO2, 2.5 mL/min, 220 nm | ≥ 99.5% | 99.8% |
| Water content | Karl Fischer coulometry (Metrohm 831) | ≤ 1.0% w/w | 0.3% |
| Residual Pd | ICP‑MS, Agilent 7800 | ≤ 10 ppm | 3 ppm |
| Residual solvents | HS‑GC‑MS per ICH Q3C | DCM ≤ 600 ppm; MeOH ≤ 3000 ppm | DCM 48 ppm, MeOH 112 ppm |
In isogenic Ba/F3 proliferation assays run in parallel, osimertinib (active metabolite AZ‑5104) yields a GI50 of 0.012 µM against the T790M single mutant but is effectively inactive against the C797S triple mutant (GI50 >5 µM), while JBJ-04-125-02 maintains a consistent GI50 of 0.50 µM irrespective of C797S status. Combination treatment with the two agents at fixed molar ratios reveals a fractional inhibitory concentration index (FICI) of 0.32 in L858R/T790M/C797S models, indicative of strong synergism because the allosteric compound binds to a non-overlapping pocket and stabilizes the inactive conformation that prevents receptor dimerization, while the covalent drug permanently blocks the ATP cleft. The synergy is lost when the allosteric inhibitor is replaced by the clinical-stage EGFR allosteric inhibitor EAI045, which lacks the 4-cyanopyridin-2-yl substituent and shows a higher off-rate from the allosteric site (koff 0.023 s−1 versus 0.003 s−1 for JBJ-04-125-02, as measured by biolayer interferometry on a FortéBio Octet RED96). Moreover, the product differentiates itself from the earlier-generation allosteric probe JBJ-01-75-09 by virtue of the 5-fluoropyridin-3-yl group, which improves oral bioavailability in mouse pharmacokinetic studies (F = 34% at 10 mg/kg p.o., compared to <5% for the des‑fluoro pyridine analog).
| Kinase | % Inhibition | Kinase | % Inhibition |
|---|---|---|---|
| EGFR (L858R/T790M/C797S) | 98 | EGFR (WT) | 12 |
| BLK | 84 (only if e.e. <99.0%) | EPHA2 | 79 (enantiomer-dependent) |
| JAK2 | 7 | ABL1 | 5 |
| SRC | 9 | KDR (VEGFR2) | 6 |
Dynamic vapor sorption (DVS) analysis performed on a Surface Measurement Systems DVS Adventure instrument at 25 °C shows that the amorphous solid adsorbs 2.8% water at 90% RH, triggering partial re-crystallization upon desorption and a 6% loss in chromatographic purity. Therefore, immediate resealing of the storage container after use and avoidance of humidity above 60% during aliquot preparation are enforced. Activity in cell-free biochemical format is diminished by pre-incubation with high protein-content media containing 10% fetal bovine serum due to albumin binding (fraction unbound 0.12), so IC50 values obtained in serum-free conditions (26 nM HTRF) should be interpreted with the understanding that physiological protein shifts the apparent potency by approximately five- to eight-fold. Published data for repeated-dose toxicology in rodents are not yet available, and the compound is classified as a hazardous substance under the OSHA Hazard Communication Standard (29 CFR 1910.1200); handling in a certified fume hood with nitrile gloves is mandatory.
Batch-to-batch consistency of the amorphous phase is monitored by modulated differential scanning calorimetry (TA Instruments Discovery DSC 250) to confirm a glass transition temperature (Tg) midpoint of 117.3 ± 1.5 °C; deviations beyond this range signal crystallinity that slows dissolution and causes variable cellular exposure. In the event that a received batch exhibits a Tg below 114 °C, the material is reconstituted in acetonitrile and re-lyophilized under controlled nucleation at −45 °C shelf temperature and 100 mTorr chamber pressure (SP Scientific VirTis AdVantage Pro). The resulting cake is micronized using a Fritsch Pulverisette 7 planetary mill with 5 mm zirconia beads at 400 rpm for 15 min under continuous nitrogen purge to prevent electrostatic charging and amorphization reversion.
Pre-formulation solubility in aqueous vehicles suitable for in vivo dosing was tested in 0.5% methylcellulose/ 0.1% Tween‑80, yielding a suspension at 2.0 mg/mL that maintains chemical integrity for 4 h at room temperature. For intravenous pharmacokinetic profiling, a clear solution at 1.0 mg/mL is obtained in 10% DMSO/ 20% Kolliphor EL/ 70% saline and remains free of visible precipitation for 24 h under gentle agitation. These formulation windows are narrower than those of the ATP-competitive inhibitor osimertinib, which dissolves at 10 mg/mL in the same vehicle, reflecting the higher crystallinity of its mesylate salt; the amorphous nature of JBJ-04-125-02 is both a benefit for dissolution rate and a liability for physical stability.
