Motolimod (VTX-378) (VTX-2337) is an effective and specific Toll-like receptor (TLR) 8 agonist (EC50: 100 nM), > 50-fold selectivity over TLR7.

TLR8:100 nM(EC50)

The activity of specific TLR agonists is assessed using the secretory embryonic alkaline phosphatase (SEAP) reporter gene that is linked to NF-κB activation in response to TLR stimulation. Measurement of SEAP activity using the Quanti-blue substrate (InvivoGen) after TLR agonist treatment is carried out.

PBMCs or purified NK cells are prepared as previously described, and the purity of NK cells was approximately 99%. NK cell-mediated cytotoxicity is assessed by Calcein-AM release from labeled target cells. In brief, PBMCs or purified NK cells are cultured for 48 hours in RPMI medium in the presence of VTX-2337 (167 or 500 nmol/L) before incubation with target cells.

VTX-378, VTX-2337

H2O: < 1 mg/mL (insoluble or slightly soluble)

Ethanol: 12 mg/mL (26.17 mM), Sonication is recommended.

DMSO: 10.71 mg/mL (23.35 mM), Sonication is recommended.

10% DMSO+90% Saline: 1.07 mg/mL (2.33 mM), Solution.

10% DMSO+40% PEG300+5% Tween 80+45% Saline: 2 mg/mL (4.36 mM), Sonication is recommended.