GW3965 hydrochloride

GW3965 hydrochloride (GW3965 HCl) belongs to synthetic small molecules and is a liver X receptor (LXR) agonist (hLXRα EC50 = 190 nM; hLXRβ EC50 = 30 nM) with subtype selectivity, cell permeability, and oral activity. This compound is used in research related to metabolism, inflammation, and neuropathic pain.

LXRα (human):190 nM (EC50), LXRβ (human):30 nM (EC50)

Methods: Spleen-derived myeloid-derived suppressor cells (MDSCs) from septic mice were incubated with 1 μM GW3965 hydrochloride for 1 hour, and cell apoptosis was detected by flow cytometry.
Results: GW3965 hydrochloride induced apoptosis of spleen MDSCs. [1]
Methods: Gefitinib-resistant non-small cell lung cancer PC9 cells were treated with 5 μM GW3965 hydrochloride alone or in combination with gefitinib for 48 hours. Cell viability was detected by MTT assay, autophagy-related proteins LC3 II/I and Beclin 1 expression were determined by Western blot, autophagosome formation was observed by AO staining, and cell apoptosis was evaluated by flow cytometry.
Results: GW3965 hydrochloride combined with gefitinib synergistically inhibited cell viability compared to monotherapy groups, upregulated LC3 II/I ratio and Beclin 1 expression, and increased autophagosome accumulation and apoptosis rate. [2]

Methods: A collagenase-induced intracerebral hemorrhage mouse model was used. GW3965 hydrochloride was administered by intraperitoneal injection at a dose of 10 mg/kg, dissolved in 50% DMSO, starting 1 hour post-surgery, once daily for 7 consecutive days.
Results: The GW3965 hydrochloride treatment group showed reduced lesion volume, accelerated hematoma clearance, alleviated white matter injury, and promoted neurological functional recovery.[3]
Methods: A cecal ligation and puncture (CLP)-induced sepsis mouse model was used. GW3965 hydrochloride was administered by subcutaneous injection at a dose of 3 mg/kg, dissolved in 5% DMSO/30% PEG300/5% Tween 80/60% ddH₂O, for a total of 6 doses at 1, 6, 12, 24, 48, and 72 hours post-surgery.
Results: GW3965 hydrochloride improved mouse survival rate, alleviated multi-organ injury, and reduced serum inflammatory factor levels.[4]

Steady-state drug accumulation assay: AuxB1 and CHrB30 cells are grown to confluency in 12-well (24 mm) tissue culture dishes and the steady-state accumulation of [3H]-vinblastine is measured. Accumulation is initiated by the addition of 0.1 μ Ci [3H]-vinblastine and unlabelled vinblastine to a final concentration of 100 nM . The accumulation of [3H]-paclitaxel is measured using 0.1 μ Ci [3H]-paclitaxel and unlabelled drug to a final concentration of 1 μM . Cells are incubated in a reaction volume of 1 mL for 60 min at 37 ℃ under 5% CO2 in order to reach steady-state. The effect of the modulators XR9576 on [3H]-ligand accumulation is investigated in the concentration range 10-9 - 10-6 M. Modulators are added from a DMSO stock giving a final solvent concentration of 0.2 % (v/v). Following cell harvesting, accumulated drug is measured by liquid scintillation counting and normalized for cell protein content. Plots of amount accumulated as a function of modulator concentration are fitted with the general dose-response equation: Y={(a-b)/(1+(X/c)d)}+b Where: Y=response; a=initial response; b=final response; c=EC50 concentration; d=slope value; X=drug concentration.

Cells are seeded in 96 wells and are treated after 24 hours with different drugs indicated in each experiment in medium containing 1% FBS or lipoprotein deficient serum. Relative proliferation is determined using Cell Proliferation Assay Kit. Cells are incubated 1.5 hrs after adding tetrazolium salt WST-1 [2-(4-iodophenyl)-3- (4-nitrophenyl)-5-(2, 4-disulfo-phenyl)-2H-tetrazolium, monosodium salt] at 5% CO2, 37oC and the absorbance of the treated and untreated cells are measured using a microplate reader at 420 to 480 nm. Cells seeded in 12 well plates are counted using a hemocytometer, and dead cells are assessed using trypan blue exclusion assays.

DMSO: 71.2 mg/mL (115.12 mM), Sonication is recommended.

Ethanol: 12.4 mg/mL (20.05 mM), Sonication is recommended.

10% DMSO+40% PEG300+5% Tween 80+45% Saline: 2 mg/mL (3.23 mM), Sonication is recommended.