| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
TLR8 agonist 5 specifically targets Toll-like receptor 8 (TLR8), an endosomal pattern recognition receptor expressed primarily on myeloid cells such as monocytes, macrophages, and dendritic cells. Activation of TLR8 triggers the MyD88-dependent signaling pathway, leading to the production of pro-inflammatory cytokines (TNF-alpha, IL-12) and type I interferons (IFN-alpha). This shifts the immune response toward a Th1-dominant profile.
|
|---|---|
| ln Vitro |
TLR8 agonist 5 is a potent TLR8 agonist/activator with an EC50 of 20 nM for HEK-Blue hTLR8. This indicates high potency for activating the TLR8 receptor. TLR8 agonist 5 activates the immune response. The compound induces the production of pro-inflammatory cytokines and type I interferons in TLR8-expressing cells. Specific cellular activation data is not detailed, but it is designed to stimulate robust immune activation.
|
| ln Vivo |
Specific in vivo activity data for TLR8 agonist 5 has not been published. As a potent TLR8 agonist (EC50 = 20 nM), it is predicted to activate innate immune responses and promote Th1-type adaptive immunity. It may have applications as a vaccine adjuvant to enhance antigen-specific immune responses, and as a monotherapy for cancer immunotherapy by remodeling the tumor microenvironment to promote anti-tumor immunity.
|
| Enzyme Assay |
The specific in vitro protocol for TLR8 activation uses a HEK-Blue™ TLR8 reporter cell assay. HEK-Blue hTLR8 cells (which express a secreted embryonic alkaline phosphatase (SEAP) reporter gene under the control of NF-kappaB and AP-1 promoters) are seeded in 96-well plates at 5×10⁴ cells/well. TLR8 agonist 5 is serially diluted (0.01 nM to 10 uM) and added to the cells. After 18-24 hours of incubation at 37degC, 20 uL of cell supernatant is transferred to a new 96-well plate and mixed with 180 uL of QUANTI-Blue™ substrate. The plate is incubated for 1-2 hours at 37degC, and SEAP activity is measured by absorbance at 620 nm. The EC50 value (20 nM) is calculated by fitting the dose-response curve.
|
| Cell Assay |
For in vitro cellular assays, primary human PBMCs are isolated from healthy donors. PBMCs are seeded in 96-well plates at 2×10⁵ cells/well. TLR8 agonist 5 is added at concentrations ranging from 1 nM to 1 uM. After 24-48 hours of incubation at 37degC, cell culture supernatants are collected. Cytokine levels (TNF-alpha, IL-12p70, IL-1beta, IFN-alpha) are quantified using a multiplex Luminex assay or ELISA. For DC maturation assays, human monocyte-derived dendritic cells (moDCs) are generated and treated with TLR8 agonist 5 for 24 hours, followed by flow cytometry analysis of CD80, CD83, CD86, and HLA-DR expression.
|
| Animal Protocol |
An in vivo protocol for TLR8 agonist 5 would involve a CT26 syngeneic mouse model, noting that TLR8 is not functional in mice (mouse TLR8 is non-functional). Therefore, human TLR8 transgenic mice or non-human primates are required. In a human TLR8 transgenic mouse model, animals are implanted subcutaneously with CT26 cells. When tumors reach 50-100 mm3, TLR8 agonist 5 is administered intratumorally or intravenously at doses of 0.1-5 mg/kg on days 7, 10, and 14. Tumor volumes are measured with calipers. At termination, tumors are collected for flow cytometry analysis of myeloid cell infiltration and T-cell activation. Cytokine levels in plasma are measured by ELISA.
|
| ADME/Pharmacokinetics |
Detailed pharmacokinetic data for TLR8 agonist 5 is not available. As a small molecule TLR8 agonist, a standard PK study in non-human primates would be conducted due to species specificity. The compound would be administered intravenously and intratumorally to determine its half-life, clearance, and tissue distribution. For research applications, it is typically formulated in PBS or saline for injection.
|
| Toxicity/Toxicokinetics |
Toxicology data for TLR8 agonist 5 is not available. As a TLR8 agonist, the primary safety concern is systemic inflammation following over-activation of the innate immune system, potentially leading to cytokine release syndrome (CRS) characterized by fever, hypotension, and multi-organ dysfunction. Standard preclinical safety assessment would include a single-dose toxicity study in cynomolgus monkeys (since TLR8 is functional in primates) to determine the MTD and monitor cytokine levels.
|
| References | |
| Additional Infomation |
TLR8 agonist 5 is a research-grade chemical and is not approved for clinical use. Its molecular formula is C31H40N6O5 with a molecular weight of 576.69. It is a potent TLR8 agonist with an EC50 value of 20 nM for HEK-Blue hTLR8. TLR8 agonist 5 activates the immune response. It is used in immunology and cancer research to study TLR8-dependent immune activation. The compound is stored at -20degC and is soluble in DMSO. Note that mouse TLR8 is non-functional, so human TLR8 transgenic models or primate studies are required for in vivo work.
|
| Molecular Formula |
C31H40N6O5
|
|---|---|
| Molecular Weight |
576.686507225037
|
| Exact Mass |
576.306
|
| CAS # |
2413016-41-6
|
| PubChem CID |
146490380
|
| Appearance |
White to off-white solid powder
|
| LogP |
3
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
14
|
| Heavy Atom Count |
42
|
| Complexity |
882
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O(CCNC(C(C)(C)NC(=O)OCC1C=CC=CC=1)=O)C(C)(C)CN1C(COCC)=NC2C(N)=NC3C=CC=CC=3C1=2
|
| InChi Key |
UXDUNZGDEXDOEZ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C31H40N6O5/c1-6-40-19-24-35-25-26(22-14-10-11-15-23(22)34-27(25)32)37(24)20-30(2,3)42-17-16-33-28(38)31(4,5)36-29(39)41-18-21-12-8-7-9-13-21/h7-15H,6,16-20H2,1-5H3,(H2,32,34)(H,33,38)(H,36,39)
|
| Chemical Name |
benzyl N-[1-[2-[1-[4-amino-2-(ethoxymethyl)imidazo[4,5-c]quinolin-1-yl]-2-methylpropan-2-yl]oxyethylamino]-2-methyl-1-oxopropan-2-yl]carbamate
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO: 50 mg/mL (86.70 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.34 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 (4.34 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 (4.34 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 | 1.7340 mL | 8.6702 mL | 17.3403 mL | |
| 5 mM | 0.3468 mL | 1.7340 mL | 3.4681 mL | |
| 10 mM | 0.1734 mL | 0.8670 mL | 1.7340 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
