| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
CU-CPT-9a targets human Toll‑like receptor 8 (TLR8). IC50 = 0.5 ± 0.1 nM in HEK‑Blue TLR8 cells (SEAP reporter assay) [1].
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| ln Vitro |
CU-CPT-9a is a selective TLR8 antagonist with an IC50 of 0.5±0.1 nM. The rise in downstream protein levels generated by R848 could be reversed by CU-CPT-9a in a dose-dependent manner. In contrast, expression of TRIF and IRF3 (cytoplasmic and nuclear) was only responsive to TLR4 and TLR3 and not to TLR837. The expression levels of TRIF and IRF3 did not reveal significant changes in THP-1 cells following R848 therapy, nor did they change after CU-CPT-9a treatment. Both CU-CPT8m and CU-CPT-9a strongly decreased TNF-α levels in a dose-dependent manner, which is consistent with prior observations on the involvement of TLR8 in these autoimmune disorders [1].
CU-CPT-9a potently and selectively inhibited TLR8‑mediated NF‑κB activation with an IC50 of 0.5 ± 0.1 nM in HEK‑Blue TLR8 cells, showing no significant inhibition of other TLRs (TLR1/2, TLR2/6, TLR3, TLR4, TLR5, TLR7, TLR9) at concentrations up to 1 μM [1]. In THP‑1 cells, CU-CPT-9a dose‑dependently reversed R848‑induced elevation of downstream signaling proteins: phosphorylated IRAK‑4 (p‑IRAK4), TRAF3, and the p65 subunit of NF‑κB, as shown by immunoblot analysis [1]. In PBMCs from rheumatoid arthritis patients, CU-CPT-9a significantly suppressed R848‑induced TNF‑α production in a dose‑dependent manner (0, 2.5, 10, 20, 40 μM) [1]. |
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| ln Vivo |
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| Cell Assay |
TLR8 inhibitory activity was measured in HEK‑Blue TLR8 cells using a SEAP reporter assay. Cells (3.5 × 10⁵ cells/mL) were treated with R848 (1 μg/mL) and varying concentrations of CU-CPT-9a for 20‑24 h. SEAP activity in the culture supernatant was detected with Quanti‑Blue reagent by absorbance at 620 nm. IC50 was calculated from dose‑response curves [1].
Immunoblot analysis: THP‑1 cells or HEK‑Blue TLR8 cells were treated with R848 and CU-CPT-9a. After lysis, proteins were fractionated into cytoplasmic and nuclear fractions, separated by SDS‑PAGE, transferred to nitrocellulose or PVDF membranes, and probed with antibodies against p‑IRAK4, IRAK4, TRAF3, IRF3, TRIF, p65, β‑actin, GAPDH, and lamin A/C. Visualization was performed with chemiluminescent substrates [1]. PBMC isolation and TNF‑α ELISA: Human PBMCs from RA patients were isolated by density gradient centrifugation, cultured at 3 × 10⁶ cells/mL in 96‑well plates, and treated with CU-CPT-9a (0, 2.5, 10, 20, 40 μM) together with R848 (1 μg/mL) for 24 h. Supernatants were collected and TNF‑α levels measured by human TNF‑α OptEIA ELISA kit [1]. |
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| Animal Protocol |
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| References | |||
| Additional Infomation |
CU-CPT-9a is a 4‑phenyl‑1‑(2H)-phthalazinone derivative. It binds to a novel allosteric site on the TLR8 dimer interface, distinct from the agonist binding site (Site 1), stabilizing the pre‑formed dimer in its resting state and preventing the conformational change required for activation. This mechanism was validated by X‑ray crystallography for the analog CU‑CPT9b (PDB 5WYZ). CU-CPT-9a demonstrated negligible cytotoxicity up to 100 μM in HEK‑Blue TLR8 cells and was effective in suppressing spontaneous TNF‑α and IL‑1β production from synovial membrane cultures of osteoarthritis patients and from PBMCs of rheumatoid arthritis and adult‑onset Still’s disease patients [1].
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| Molecular Formula |
C17H15NO2
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| Molecular Weight |
265.306504487991
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| Exact Mass |
265.11
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| CAS # |
2165340-32-7
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| Related CAS # |
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| PubChem CID |
135567383
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.8
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
20
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| Complexity |
322
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O(C)C1C=CC2C(C=1)=NC=CC=2C1C=CC(=C(C)C=1)O
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| InChi Key |
HNYBTVKYLVLWCB-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C17H15NO2/c1-11-9-12(3-6-17(11)19)14-7-8-18-16-10-13(20-2)4-5-15(14)16/h3-10,19H,1-2H3
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| Chemical Name |
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| Synonyms |
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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 |
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| 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) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (9.42 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 (9.42 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 (9.42 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 | 3.7692 mL | 18.8459 mL | 37.6918 mL | |
| 5 mM | 0.7538 mL | 3.7692 mL | 7.5384 mL | |
| 10 mM | 0.3769 mL | 1.8846 mL | 3.7692 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.
 CU-CPT8m potently and selectively inhibited TLR8. Crystal structure of the TLR8/CU-CPT8m complex. |
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 TLR8 inhibitors suppress the proinflammatory cytokine production in multiple human primary cells derived from different patients. |
 Proposed antagonistic mechanism of CU-CPT compounds (top) and schematic representation of domain arrangement in each TLR8 forms (bottom). TLR8 inhibitors consistently recognize an allosteric pocket on the protein-protein interface, stabilizing the inactive TLR8 dimer |
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