| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| Other Sizes |
| Targets |
PRMT5 (binds to the PRMT5-PBM interface; covalent inhibitor targeting Cys278; FP competition IC50 at 40 min = 13.8 μM; kinact/KI = 244 M⁻¹sec⁻¹; Kinact = 0.715 min⁻¹; KI = 49 μM) [1].
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| ln Vitro |
BRD0639 showed time-dependent displacement of a fluorescently-labeled RIOK1 PBM peptide from PRMT5:WDR77 complex in a fluorescence polarization (FP) competition assay, with IC50 decreasing from >100 μM at early time points to 13.8 μM at 40 min [1].
In a GSH reactivity assay, BRD0639 had a half-life (T½) of 916 min, indicating low intrinsic reactivity compared to dichloro analogs (e.g., compound 6 T½ 31 min) [1]. Kinetic analysis by intact mass spectrometry (kinact/KI assay) gave a kinact/KI ratio of 244 M⁻¹sec⁻¹, with kinact = 0.715 min⁻¹ and KI = 49 μM [1]. In a PRMT5 C278A mutant, BRD0639 showed no activity in FP competition (IC50 > 100 μM vs 0.6 μM for wild-type), confirming covalent binding to Cys278 [1]. Surface plasmon resonance (SPR) showed that pre-incubation of PRMT5 with BRD0639 greatly reduced binding to full-length pICln (KD 32 nM for DMSO-treated vs non-specific binding for compound-treated), indicating irreversible disruption of PRMT5-pICln interaction [1]. Cryo-EM structure (PDB 7M05) of a related analog (compound 6) bound to PRMT5 revealed the binding mode; BRD0639 is expected to bind similarly with the chloropyridazine forming a covalent bond to Cys278 and the pyridylethyl side chain folded back to create pi-pi stacking with Tyr286 and Phe243 [1]. |
| Enzyme Assay |
Fluorescence polarization (FP) competition assay: 200 nM PRMT5:WDR77 protomer, 10 nM KU560-labeled RIOK1-derived PBM peptide, 50 mM HEPES pH 7.4, 100 mM NaCl, 0.5 mM TCEP, 0.01% Tween 20. Compounds were serially diluted, incubated for 40 min at room temperature, and FP measured using a plate reader (Rhodamine or Bodipy TMR filter set). IC50 values were calculated [1].
GSH reactivity assay: 10 μM compound was incubated with 5 mM glutathione (GSH) in PBS pH 7.4 at 37°C. At various time points (0-1440 min), aliquots were quenched with cold acetonitrile containing internal standard, and parent compound remaining was quantified by LC-MS/MS. Half-life (T½) was calculated using first-order kinetics [1]. Kinact/KI assay by intact mass spectrometry: Purified PRMT5:WDR77 (50 nM) was incubated with various concentrations of compound (12-point dilution) in reaction buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 1 mM TCEP). At time points (0-60 min), aliquots were quenched with 0.5% formic acid. Unmodified PRMT5 was quantified by LC-MS/MS using MRM (multiple reaction monitoring) of specific charge states (m/z 855.0→854.9 for PRMT5, z=85). The fraction unmodified was plotted vs time, and kinact and KI were determined by fitting to the equation for irreversible inactivation [1]. Surface plasmon resonance (SPR): PRMT5 was pre-incubated with 2 μM BRD0639 or DMSO overnight, then immobilized on a chip. Full-length pICln was titrated as analyte (0-1000 nM). Binding responses were fitted to a one-site total and non-specific binding model [1]. Cryo-EM: PRMT5:WDR77 complex (5 μM) was co-incubated with 25 μM compound 6 (a close analog) and 20 μM JNJ-64619178 (catalytic inhibitor) overnight. Complex was purified by size exclusion, concentrated to 10 mg/ml, and applied to grids. Data were collected on a Titan Krios microscope with K2 detector. 2169 movies were processed in cisTEM, yielding a 2.39 Å resolution map (PDB 7M05) [1]. |
| Cell Assay |
Target engagement assay in Expi293 cells: Cells overexpressing HA-tagged WDR77 and untagged PRMT5 were treated with BRD0639 at indicated concentrations (0-30 μM) for 6 h. PRMT5 complex was immunoprecipitated with anti-HA agarose, eluted with HA peptide, and analyzed by intact mass LC-MS to quantify PRMT5-adduct formation. EC50 for adduct formation was approximately 3 μM, with maximal ~40% labeling [1].
NanoBiT PRMT5-RIOK1 complementation assay in permeabilized cells: HEK293T cells stably expressing SmBiT-PRMT5 and LgBiT-RIOK1 were treated with compounds for 40 min, then lysed with buffer (10% glycerol, 50 mM Tris-HCl, 150 mM KCl, 2 mM EDTA, 0.1% NP40). Nano-Glo substrate was added and luminescence measured. IC50 for BRD0639 was 7.5 μM [1]. NanoBiT assay in intact cells: Same cells were treated with compounds for 40 min in culture medium, then Nano-Glo Live Cell Substrate was added and luminescence measured. IC50 for BRD0639 was 16 μM, compared to inactive analog 30 (IC50 >100 μM) [1]. Western blot for symmetric arginine dimethylation (SDMA): HCT116 MTAP-/- cells were treated with 25 μM BRD0639 for 24 h (with media refresh and retreatment at 12 h). Lysates were blotted with anti-SDMA antibody (CST13222). BRD0639 reduced SDMA levels on a subset of proteins, recapitulating the effect of genetic disruption of the PBM binding site (PRMT5 ADA mutant) [1]. Control experiments with inactive analog 30 (compound 30, a 5-chloro regioisomer) showed no effect on SDMA, confirming on-target activity [1]. |
| Toxicity/Toxicokinetics |
In vitro GSH reactivity: BRD0639 had a half-life of 916 min in 5 mM GSH, indicating low electrophilic reactivity compared to dichloro analogs (e.g., compound 6 T½ 31 min). No acute toxicity data reported [1].
No in vivo toxicity data available [1]. |
| References | |
| Additional Infomation |
BRD0639 is a first-in-class, covalent, PBM-competitive inhibitor of PRMT5. It selectively targets the protein-protein interaction between PRMT5 and its substrate adaptor proteins (pICln, Riok1) by binding to a surface groove distal to the catalytic site. The compound forms a covalent bond with Cys278 of PRMT5 via a monochloropyridazine warhead after initial non-covalent binding. Optimization from a dichloropyridazine hit (compound 1) reduced intrinsic reactivity (GSH t½ from 31 min to 916 min) while maintaining potency (FP IC50 13.8 μM). BRD0639 disrupts PRMT5-RIOK1 complex in cells (IC50 7.5-16 μM) and reduces symmetric dimethylation of a subset of substrates, phenocopying genetic disruption of the PBM binding site. Unlike catalytic PRMT5 inhibitors that block all substrates, BRD0639 selectively impairs PBM-dependent methylation, particularly affecting splicing-related functions. This compound serves as a chemical probe to study PBM-dependent PRMT5 activities and may inform development of non-catalytic PRMT5 inhibitors. No FDA approval or clinical trial status is mentioned [1].
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| Appearance |
Typically exists as solid at room temperature
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| Chemical Name |
(S)-2-(4-chloro-6-oxopyridazin-1(6H)-yl)-N-(4-methyl-3-(N-(2-(pyridin-2-yl)ethyl)sulfamoyl)phenyl)propanamide
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| Synonyms |
BRD 0639BRD0639 BRD-0639
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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