CL097 is a potent TLR7 and TLR8 agonist that induces pro-inflammatory cytokines in macrophages. CL097 induces NADPH oxidase priming activity, resulting in increased fMLF-stimulated ROS production.

Physicochemical Properties

Molecular Formula	C13H14N4O
Molecular Weight	242.28
Exact Mass	242.116
CAS #	1026249-18-2
Related CAS #	CL097 hydrochloride
PubChem CID	11579618
Appearance	White to off-white solid powder
LogP	1.5
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	3
Heavy Atom Count	18
Complexity	288
Defined Atom Stereocenter Count	0
SMILES	N1C2C(=CC=CC=2)C2N=C(COCC)NC=2C=1N
InChi Key	DEVCLHVFELRPIU-UHFFFAOYSA-N
InChi Code	InChI=1S/C13H14N4O/c1-2-18-7-10-16-11-8-5-3-4-6-9(8)15-13(14)12(11)17-10/h3-6H,2,7H2,1H3,(H2,14,15)(H,16,17)
Chemical Name	2-(ethoxymethyl)-3H-imidazo[4,5-c]quinolin-4-amine
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
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro	CL097 activates NF-κB at 0.1 μM in TLR7-transfected HEK293 cells and at 4 μM in TLR8-transfected HEK293 cells [1]. By imitating the phosphorylation of p47phox in the human neutrophil signaling system, CL097 causes overactivation of NADPH oxidase, suggesting that p38 MAPK, ERK1/2, protein inhibitor C, and Pin1 regulate this process. CL097 increases fmlf-induced p47phox and causes phosphorylation of p47phox at particular locations.
ln Vivo	CL097 and CD40 agonists promote significant diabetogenic cytotoxic T pathway (CTL) activity in NOD mice. CL097 (5 mg/kg, sc) alone induced possible excess (~25%) of the target peptide. However, combination treatment with CL097 and a CD40 agonist (10 mg/kg, ip) resulted in an approximately twofold increase in the oscillation frequency of IGRP peptide-coated targets compared to treatment with CL097 alone [3].
Cell Assay	Western Blot analysis [2] Cell Types: Neutrophil Tested Concentrations: 0, 0.5, 2.5, 5 and 10 μg/mL Incubation Duration: 30 minutes of pretreatment Experimental Results: Induces phosphorylation of p47phox at specific sites at a certain concentration [2]. Dependence method.
Animal Protocol	Animal/Disease Models: Female 8.3 NOD mice (5-6 weeks old) [3] Doses: 5 mg/kg Route of Administration: subcutaneous injection Experimental Results: Caused moderate specific cleavage of the target peptide (∼25%).
References	[1]. Agonist and antagonist ligands of toll-like receptors 7 and 8: Ingenious tools for therapeutic purposes. Eur J Med Chem. 2020 May 1;193:112238. [2]. The TLR7/8 agonist CL097 primes N-formyl-methionyl-leucyl-phenylalanine-stimulated NADPH oxidase activation in human neutrophils: critical role of p47phox phosphorylation and the proline isomerase Pin1. J Immunol. 2012 Nov 1;189(9):4657-65. [3]. Toll-like receptor 7 stimulation promotes autoimmune diabetes in the NOD mouse. Diabetologia. 2011 Jun;54(6):1407-16.

Solubility Data

Solubility (In Vitro)

DMSO : ~100 mg/mL (~412.75 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (10.32 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 (10.32 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (10.32 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	4.1275 mL	20.6373 mL	41.2746 mL
	5 mM	0.8255 mL	4.1275 mL	8.2549 mL
	10 mM	0.4127 mL	2.0637 mL	4.1275 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.