| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
CAD204520 directly binds to SERCA, specifically to the transmembrane interface between helices M1, M2, M3, and M4 of the SERCA ATPase, with an IC50 of 0.34 microM. Unlike classical SERCA inhibitors such as thapsigargin, CAD204520 minimally alters calcium shift and fails to trigger the unfolded protein response. It preferentially impairs the maturation and activity of oncogenic NOTCH1 mutants over wild-type NOTCH1 by causing a defect in NOTCH1 trafficking.
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| ln Vitro |
CAD204520 impairs the proliferation of T-ALL cell lines carrying activating mutations of NOTCH1, including PEST domain deletions. Clinical samples carrying NOTCH1 mutations are more sensitive to CAD204520 compared to normal lymphocytes or wild-type NOTCH1 ALL cells. Mechanistically, CAD204520 treatment reduces the levels of the activated form of NOTCH1. Treatment results in a significant 56-fold reduction of the percentage of circulating leukemic cells.
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| ln Vivo |
In a T-ALL xenografted mouse model, CAD204520 reduces circulating and tissue-infiltrating human leukemia T-ALL cells. Administration of 30 mg/kg BID for 21 days is well-tolerated in CD1 mice without causing loss of weight, cardiac toxicity, or gastrointestinal toxicities. CAD204520 shows no heart-related toxicity in the xenograft model, differentiating it from other SERCA inhibitors that have narrow therapeutic windows.
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| Enzyme Assay |
Surface plasmon resonance (SPR) assay using purified recombinant human SERCA protein immobilized on a CM5 chip is performed. Running buffer contains 50 mM HEPES pH 7.4, 150 mM NaCl, 2 mM CaCl2, and 0.05% Tween-20. Test compound at concentrations of 0.1-100 microM is injected at flow rate 30 microL/min for 120 seconds, followed by 300 seconds dissociation. KD is calculated using Biacore evaluation software.
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| Cell Assay |
T-ALL cell lines (e.g., SKW-3, KE-37, DND-41) carrying activating NOTCH1 mutations are seeded in 96-well plates at 5×103 cells/well and treated with CAD204520 (0.1-50 microM) for 72 hours. Cell viability is measured by MTT or CellTiter-Glo assay. NOTCH1 activation (cleaved NOTCH1 intracellular domain) and downstream target gene expression (e.g., HES1, c-MYC) are quantified by Western blot and qRT-PCR after 6-24 hour treatments.
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| Animal Protocol |
NOD/SCID mice (6-8 weeks) are injected intravenously with 5×10⁶ SKW-3 or KE-37 T-ALL cells. After engraftment (day 7), mice are randomized and treated orally with vehicle or CAD204520 at 30 mg/kg twice daily for 21 days. Circulating leukemic cells are measured by flow cytometry using human CD45 antibody. Tissue infiltration in spleen, bone marrow, and liver is assessed by human CD45 immunohistochemistry. Cardiac function is monitored by echocardiography.
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| ADME/Pharmacokinetics |
Following oral administration, CAD204520 exhibits favorable oral bioavailability. Based on its mechanism, the compound is designed to avoid the rapid metabolic degradation and toxicity typical of thapsigargin. It demonstrates a plasma half-life suitable for BID dosing, with exposure levels sufficient to sustain NOTCH1 inhibition in target tissues.
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| Toxicity/Toxicokinetics |
Preclinical toxicology in CD1 mice shows that CAD204520 is well-tolerated at 30 mg/kg BID for 21 days with no weight loss, cardiac toxicity, or gastrointestinal toxicities. Off-target toxicity is minimal and tolerable, supporting further development for NOTCH1-dependent tumors. Somatic hotspot mutations in the SERCA2 ATPase pocket that confer resistance to known SERCA modulators do not interfere with CAD204520 binding, suggesting its activity will be unaffected by recurrent resistant genetic variants.
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| References | |
| Additional Infomation |
The compound has CAS number 2682205-21-4. Its molecular formula is C23H34Cl2F3N3O2 and molecular weight is 512.44. It is supplied as the dihydrochloride salt. Crystal structure analysis shows CAD204520 binds the transmembrane interface of SERCA between helices M1, M2, M3, and M4.
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| Molecular Formula |
C23H34CL2F3N3O2
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|---|---|
| Molecular Weight |
512.44
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| Exact Mass |
511.198
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| CAS # |
2682205-21-4
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| Related CAS # |
CAD204520
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| PubChem CID |
165789190
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
3
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
33
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| Complexity |
559
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCCC1=C(NC2=C1C=CC(=C2)OC(F)(F)F)C3CCCCN3CCN4CCOCC4.Cl.Cl
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| InChi Key |
OTJSFNAMCJKNOR-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C23H32F3N3O2.2ClH/c1-2-5-19-18-8-7-17(31-23(24,25)26)16-20(18)27-22(19)21-6-3-4-9-29(21)11-10-28-12-14-30-15-13-28;;/h7-8,16,21,27H,2-6,9-15H2,1H3;2*1H
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| Chemical Name |
4-[2-[2-[3-propyl-6-(trifluoromethoxy)-1H-indol-2-yl]piperidin-1-yl]ethyl]morpholine;dihydrochloride
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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. |
| 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9514 mL | 9.7572 mL | 19.5145 mL | |
| 5 mM | 0.3903 mL | 1.9514 mL | 3.9029 mL | |
| 10 mM | 0.1951 mL | 0.9757 mL | 1.9514 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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