| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
TLR4 1.675 μM (IC50, in HEK293 cells)
CAY10614 specifically targets Toll-like receptor 4 (TLR4). It is a TLR4 antagonist that inhibits lipid A-induced activation of TLR4. Lipid A is the bioactive component of lipopolysaccharide (LPS), which is the natural ligand for the TLR4/MD-2 complex. By antagonizing TLR4, CAY10614 prevents the activation of the MyD88-dependent and TRIF-dependent signaling pathways, blocking the production of pro-inflammatory cytokines such as TNF-alpha, IL-6, and IL-1beta. |
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| ln Vitro |
In HEK293 cells, lipid A-induced increases in phosphatase activity are inhibited in a concentration-dependent manner by CAY10614 (Compound 7) (1–10 μM) [1]. In neurons cultured for more than 18 days in vitro (DIV), CAY10614 (0.5 μM) suppresses the rise in [Ca2+]cyt brought on by LPS [2].
In vitro, CAY10614 is a TLR4 antagonist. It inhibits lipid A-induced activation of TLR4 (IC50 = 1.675 microM). This has been determined in a cell-based assay using modified human embryonic kidney (HEK) cells. It does not significantly affect HEK cell viability, indicating low cytotoxicity at concentrations up to 10 microM. CAY10614 is a potent TLR4 antagonist and has been used as a tool compound to study TLR4 biology. |
| ln Vivo |
Mice given intraperitoneal LPS (20 mg/kg) had considerably higher survival rates when CAY10614 (Compound 7) (10 mg/kg; intraperitoneally injected 30 minutes before LPS) is administered[1].
In vivo, CAY10614 improves survival in a mouse model of lethal endotoxic shock. It also significantly improves survival of mice given intraperitoneal LPS. By blocking TLR4 activation, it reduces the systemic inflammatory response (cytokine storm) that is typically responsible for mortality in endotoxemia models. CAY10614 can be used for sepsis research to explore therapeutic strategies targeting TLR4. |
| Enzyme Assay |
The specific protocol for assessing TLR4 antagonism uses a HEK-Blue™ TLR4 reporter cell assay. HEK-Blue hTLR4 cells are seeded in 96-well plates at 5×10⁴ cells/well. CAY10614 is serially diluted (0.01 microM to 100 microM) and added to the cells. After a 1-hour pre-incubation, lipid A (from E. coli, 100 ng/mL) is added to activate TLR4. The cells are incubated for 18-24 hours at 37degC. Then, 20 microL of cell supernatant is mixed with 180 microL of QUANTI-Blue™ substrate. The plate is incubated for 1-2 hours at 37degC, and SEAP activity (as a measure of NF-kappaB activation) is measured by absorbance at 620 nm. The IC50 value of 1.675 microM is calculated by fitting the dose-response curve using a 4-parameter logistic model.
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| Cell Assay |
For in vitro cellular assays, primary human peripheral blood mononuclear cells (PBMCs) are isolated from healthy donors. PBMCs are seeded in 96-well plates at 2×10⁵ cells/well. CAY10614 is added at concentrations ranging from 0.1-50 microM and pre-incubated for 1 hour. Cells are then stimulated with LPS (E. coli, 10 ng/mL) for 24 hours. Cell culture supernatants are collected, and human TNF-alpha, IL-6, and IL-1beta are quantified by ELISA. The ability of CAY10614 to inhibit LPS-induced cytokine production is calculated as percent inhibition relative to the LPS-only control. Cell viability is assessed using the MTT assay to ensure that the effects are due to TLR4 antagonism and not cytotoxicity.
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| Animal Protocol |
Animal/Disease Models: C57BL/6J male mice (9 weeks) were ip injected with 20 mg/kg LPS[1]
Doses: 10 mg/kg Route of Administration: Ip 30 min before the LPS Experimental Results: Increased the survival rate of mice from 0% to 67%. An in vivo protocol for CAY10614 uses a mouse model of LPS-induced endotoxic shock. Female C57BL/6 mice (8-10 weeks old, n=10 per group) are injected intraperitoneally (i.p.) with a lethal dose of LPS (E. coli 0111:B4, 15-20 mg/kg). CAY10614 is dissolved in a suitable vehicle (e.g., 10% DMSO/40% PEG400/50% saline). For prophylactic treatment, CAY10614 is administered i.p. at doses of 1, 3, or 10 mg/kg, 1 hour before LPS injection. For therapeutic treatment, it can be given 1-2 hours after LPS injection. Mice are monitored for survival every 4-6 hours for 72 hours. Blood is collected from a subset of mice at 2, 4, 6, and 12 hours post-LPS for serum TNF-alpha and IL-6 measurement by ELISA. Liver and lung tissues are collected for histopathology (H&E staining) to assess inflammation. |
| ADME/Pharmacokinetics |
Detailed pharmacokinetic data for CAY10614 is not available. As a small molecule TLR4 antagonist, a standard PK study in mice would involve IV (1 mg/kg) and IP (10 mg/kg) administration. Plasma samples would be collected at multiple time points (0-24 hours) and analyzed by LC-MS/MS. Key parameters including T1/2, Cmax, AUC, clearance, and bioavailability would be calculated. The compound's half-life is likely moderate enough to allow once- or twice-daily dosing for efficacy studies.
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| Toxicity/Toxicokinetics |
Toxicology data for CAY10614 is limited. As a TLR4 antagonist, the primary safety concern is increased susceptibility to Gram-negative bacterial infections due to impaired LPS sensing. Additionally, the compound has been reported to not significantly affect HEK cell viability, suggesting low cytotoxicity. Standard preclinical safety assessment would include a 14-day repeat-dose toxicity study in mice to determine the maximum tolerated dose (MTD), and in vitro hERG channel inhibition testing to assess cardiac safety. No significant organ toxicity has been reported at efficacious doses.
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| References |
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| Additional Infomation |
CAY10614 is a research-grade chemical and is not approved for clinical use. Its molecular formula is C26H31N3O5S with a molecular weight of 497.61. It is a potent TLR4 antagonist that inhibits lipid A-induced TLR4 activation with an IC50 of 1.675 microM. It does not significantly affect HEK cell viability. CAY10614 improves survival in a mouse model of lethal endotoxic shock. This compound is a valuable tool for sepsis research and for studying the role of TLR4 in inflammatory diseases. It is stored at -20degC and is soluble in DMSO.
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| Molecular Formula |
C42H78INO2
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|---|---|
| Molecular Weight |
755.98
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| Exact Mass |
755.507
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| CAS # |
1202208-36-3
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| PubChem CID |
25223004
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| Appearance |
White to off-white solid powder
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| LogP |
10.369
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
31
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| Heavy Atom Count |
46
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| Complexity |
630
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCCCCCCCCCCCCCOC1=C(C=C(C=C1)C[N+](C)(C)C2CCCC2)OCCCCCCCCCCCCCC.[I-]
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| InChi Key |
UXOPVNPWTUWUEA-UHFFFAOYSA-M
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| InChi Code |
InChI=1S/C42H78NO2.HI/c1-5-7-9-11-13-15-17-19-21-23-25-29-35-44-41-34-33-39(38-43(3,4)40-31-27-28-32-40)37-42(41)45-36-30-26-24-22-20-18-16-14-12-10-8-6-2;/h33-34,37,40H,5-32,35-36,38H2,1-4H3;1H/q+1;/p-1
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| Chemical Name |
cyclopentyl-[[3,4-di(tetradecoxy)phenyl]methyl]-dimethylazanium;iodide
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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, 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) |
DMSO: 12.5 mg/mL (16.53 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 0.83 mg/mL (1.10 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 8.3 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: 0.83 mg/mL (1.10 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 8.3 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3228 mL | 6.6139 mL | 13.2279 mL | |
| 5 mM | 0.2646 mL | 1.3228 mL | 2.6456 mL | |
| 10 mM | 0.1323 mL | 0.6614 mL | 1.3228 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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