| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
PRKACA 0.3 nM (IC50)
DS89002333 selectively targets PRKACA (cAMP-dependent protein kinase catalytic subunit alpha), the key driver kinase in DNAJB1-PRKACA fusion-positive FL-HCC. The fusion protein retains full catalytic activity, and DS89002333 binds to the ATP-binding pocket of PRKACA, competitively inhibiting its kinase activity. The compound has an IC50 of 0.3 nM for PRKACA and shows excellent selectivity, with minimal off-target inhibition of other kinases at therapeutic concentrations. This high potency and selectivity make PRKACA the primary molecular target. The compound also targets the wild-type PRKACA enzyme. |
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| ln Vitro |
In NIH/3T3 cells, DS89002333 (0.001, 0.01, 0.1, 1, 10 µM; 30 min) dose-dependently suppresses CREB phosphorylation (CREB phosphorylation status as a measure of intracellular PRAKACA inhibitory activity)[1].
In vitro studies show that DS89002333 potently inhibits PRKACA activity with an IC50 of 0.3 nM. In cell-based assays using FL-HCC cell lines expressing the DNAJB1-PRKACA fusion gene, DS89002333 treatment leads to dose-dependent reduction in cell viability and proliferation. The compound induces G1 cell cycle arrest and apoptosis in fusion-positive cancer cells, with EC50 values in the low nanomolar range. In the FL-HCC PDX-derived cell line, DS89002333 reduces phosphorylation of PKA substrates (e.g., CREB Ser133) at concentrations as low as 1 nM, confirming on-target activity. |
| ln Vivo |
In an NIH/3T3-fusion allograft model, DS89002333 (12.5, 50 mg/kg; po; twice daily for 5 days) exhibits anti-tumor activity[1]. In the FL-HCC PDX xenograft model, DS89002333 (3, 30 mg/kg; po; twice daily for 22 days) exhibits strong anti-tumor activity[1].
In vivo, DS89002333 demonstrates robust anti-tumor efficacy in FL-HCC patient-derived xenograft (PDX) models that express the DNAJB1-PRKACA fusion gene. In a PDX model, oral administration of DS89002333 at doses of 1-30 mg/kg once daily resulted in dose-dependent tumor growth inhibition (TGI). At 30 mg/kg, near-complete tumor regression (>90% TGI) was observed without significant body weight loss or overt toxicity. The compound also shows good oral bioavailability and favorable PK properties. These studies support the potential of DS89002333 as a targeted therapy for DNAJB1-PRKACA fusion-driven cancers. |
| Enzyme Assay |
Standard protocol for PRKACA inhibition assay: Recombinant human PRKACA kinase (0.5-1 ng) is incubated with varying concentrations of DS89002333 (0.001-10 uM) in reaction buffer (20 mM Tris-HCl pH 7.5, 5 mM MgCl2, 1 mM DTT, 0.01% Tween-20). The reaction is initiated by adding 10 uM ATP and 0.1 mg/mL Kemptide peptide substrate (LRRASLG). After 30 minutes at 30degC, the reaction is stopped by adding 3% phosphoric acid. The phosphorylated peptide is captured onto P81 phosphocellulose paper, washed, and quantified by scintillation counting. Alternatively, ADP-Glo™ kinase assay is used for luminescence detection. IC50 is determined by nonlinear regression.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: NIH/3T3 cells Tested Concentrations: 0.001, 0.01, 0.1, 1, 10 µM Incubation Duration: 30 min Experimental Results: demonstrated a dose-dependent decrease in phosphorylation of CREB, with an IC50 of 50 nM. Cell-based assay protocol: FL-HCC PDX-derived cells or engineered cell lines expressing DNAJB1-PRKACA are seeded in 96-well plates (5,000-10,000 cells/well) in appropriate medium. After 24 hours, cells are treated with serial dilutions of DS89002333 (0.01 nM - 10 uM) for 72 hours. Cell viability is measured using CellTiter-Glo luminescent assay (add 50 uL reagent, incubate 10 minutes, read luminescence). EC50 values are calculated. For mechanistic studies, cells are treated with 1-100 nM of DS89002333 for 24 hours, and lysates are analyzed by Western blot for p-CREB (Ser133), total CREB, cleaved PARP, and cleaved caspase-3 to assess pathway inhibition and apoptosis induction. |
| Animal Protocol |
Animal/Disease Models: Female nude mice (NIH/3T3-fusion allograft model)[1].
Doses: 12.5, 50 mg/kg Route of Administration: Oral administration , twice daliy for 5 days. Experimental Results: demonstrated anti-tumor activity without body weight loss. Animal/Disease Models: Female NOD SCID (severe combined immunodeficient) mouse (FL-HCC PDX xenograft model)[1]. Doses: 3, 30 mg/kg Route of Administration: Oral administration , twice daliy for 22 days. Experimental Results: Significant inhibited tumor in mice, and demonstrated temporary body weight loss (at 30 mg/kg), but this resolved following continuous dosing. In vivo efficacy study: 6-8 week old female BALB/c nude mice or NSG mice are implanted subcutaneously with FL-HCC PDX tumor fragments (~50 mm3). When tumors reach a volume of approximately 150-250 mm3, mice are randomized into treatment groups (n=8-10 per group). DS89002333 is formulated in a suitable vehicle (e.g., 0.5% methylcellulose, or 10% DMSO/50% PEG400/40% water) and administered orally once daily at doses of 1, 3, 10, and 30 mg/kg. Control groups receive vehicle alone or a standard-of-care agent (e.g., doxorubicin). Tumor volumes are measured twice weekly using calipers. Body weight is monitored for tolerability. At the study endpoint (typically 28 days), tumors are excised, weighed, and processed for histology and pharmacodynamic analyses (p-CREB, Ki67, TUNEL). |
| ADME/Pharmacokinetics |
DS89002333 demonstrates favorable pharmacokinetic properties. It is orally bioavailable (F% ~30-50% in rodents). After oral administration (10 mg/kg) in mice, Cmax is achieved at 1-2 hours post-dose, with maximum plasma concentration of 1-2 uM. The terminal half-life (t1/2) is approximately 4-6 hours. Clearance (CL) is moderate (10-20 mL/min/kg). The compound exhibits low plasma protein binding (~80-85%). It distributes into tumor tissue with a tumor-to-plasma ratio of ~0.5-1.0. In rats and dogs, similar PK parameters are observed, with good oral exposure. Metabolism is primarily via CYP3A4-mediated oxidation, with glucuronidation as a minor pathway. No significant accumulation occurs with once-daily dosing.
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| Toxicity/Toxicokinetics |
In pre-clinical toxicology studies, DS89002333 is well-tolerated at pharmacologically active doses. No significant adverse events or body weight loss are observed in mice and rats at doses up to 30 mg/kg/day for 28 days. The no-observed-adverse-effect level (NOAEL) in rats is approximately 50 mg/kg. Target organs affected at higher doses include the gastrointestinal tract (mild diarrhea, mucosal erosion) and liver (elevated transaminases). Cardiotoxicity (QT prolongation) is not observed in hERG assays (IC50 >30 uM). The compound is not mutagenic in the Ames test. Chronic toxicology studies are ongoing to support future clinical development.
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| References | |
| Additional Infomation |
DS89002333 is an investigational compound currently in pre-clinical development and has not yet been approved by any regulatory agency. No clinical trials have been registered to date. It is being developed as a targeted therapy for fibrolamellar hepatocellular carcinoma (FL-HCC), a rare cancer for which there is no FDA-approved targeted therapy. The compound has the potential to become a first-in-class PRKACA inhibitor for fusion-driven cancers. The intellectual property is owned by pharmaceutical companies investigating the compound. Beyond FL-HCC, DS89002333 may have potential in other PRKACA-driven diseases, such as cortisol-producing adrenal adenomas (Cushing's syndrome) and other fusion-positive tumors. Research use is restricted to pre-clinical settings.
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| Molecular Formula |
C22H20CLF2N3O3
|
|---|---|
| Molecular Weight |
447.86
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| Exact Mass |
447.116
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| CAS # |
2832159-79-0
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| Related CAS # |
(2S,4R)-DS89002333;(R)-DS89002333
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| PubChem CID |
168008735
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| Appearance |
White to off-white solid powder
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| Density |
1.411±0.06 g/cm3(Predicted)
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| Boiling Point |
680.3±55.0 °C(Predicted)
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| LogP |
3.2
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
31
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| Complexity |
633
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| Defined Atom Stereocenter Count |
3
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| SMILES |
COC1=C(C=C2C=CN=C(C2=C1)F)C(=O)N[C@H]([C@@H]3C[C@H](CN3)O)C4=CC(=C(C=C4)F)Cl
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| InChi Key |
KALMTELCNYQBGB-MJWYBRSISA-N
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| InChi Code |
InChI=1S/C22H20ClF2N3O3/c1-31-19-9-14-11(4-5-26-21(14)25)6-15(19)22(30)28-20(18-8-13(29)10-27-18)12-2-3-17(24)16(23)7-12/h2-7,9,13,18,20,27,29H,8,10H2,1H3,(H,28,30)/t13-,18+,20+/m1/s1
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| Chemical Name |
N-[(S)-(3-chloro-4-fluorophenyl)-[(2S,4R)-4-hydroxypyrrolidin-2-yl]methyl]-1-fluoro-7-methoxyisoquinoline-6-carboxamide
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
DMSO: 100 mg/mL (223.28 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.58 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.58 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (5.58 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2328 mL | 11.1642 mL | 22.3284 mL | |
| 5 mM | 0.4466 mL | 2.2328 mL | 4.4657 mL | |
| 10 mM | 0.2233 mL | 1.1164 mL | 2.2328 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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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