GW3965 hydrochloride

Biological Activity

GW3965 hydrochloride is an agonist for the liver X receptor (LXR), reduces angiotensin II-mediated pressor responses in Sprague-Dawley rats. GW3965 acts as a full agonist of hLXRα and hLXRβ (EC50 values are 190 and 30 nM respectively) in cell-based reporter gene assays. It is orally active in mice. When screened against a panel of nuclear receptors, it cross-reacted with only the pregnane X receptor (PXR). GW3965 hydrochloride displays potent antiatherogenic activity in mouse models of atherosclerosis. GW3965 dosing blunted the vasopressor effect of Ang II, which was significantly different with the 0.3 and 3 microg kg(-1) doses. GW3965 decreased Ang II-mediated vasopressor responses coincident with a trend toward reduced ATR gene expression, suggesting that LXR agonists could affect vascular reactivity. Alternate studies suggest that GW3965 hydrochloride increases apoA-I protein levels in the central nervous system independent of ABCA1. Since ABCA1 is a major regulator of cholesterol and phospholipid metabolism, GW3965 hydrochloride is a useful tool to study these cellular functions.

Product Citations

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  Cell Death Dis. 2019 Jun 5;10(6):439.

  Loss of transglutaminase 2 sensitizes for diet-induced obesity-related inflammation and insulin resistance due to enhanced macrophage c-Src signaling.
  GW3965 hydrochloride purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Allergol Int (2015). Figure 2. GW3965
	Method	Western Blot
	Cell Lines	mice
	Concentrations	1 μM
	Incubation Time	1 h
	Results	We examined the effects of GW3965 on the IgEþAg- or LPS-induced protein expression of IL- 1a, IL-1b, IL-6, and TNF-a. Consistent with our microarray data, GW3965 significantly attenuated the protein expression of IL-1a and IL-1b, but not of IL-6 or TNF-a, in MCs stimulated by IgEþAg

Protocol

Cell Experiment
Cell lines	RAW264.7 cells
Preparation method	Cellular Internalization of LXR Agonist RAW264.7 cells were seeded into 24-well plates at a density of 1 × 105 cells per well 24 hours prior to the experiment. NPs encapsulating a GW3965 concentration of 1 μM were then incubated with cells for the desired time points. Afterwards, the cells were washed with PBS, followed by trypsinization and centrifugation to remove excess trypsin-EDTA. Cells were then resuspended in PBS, and cellular uptake of NPs was assessed via flow cytometry (BD LSR II, BD Bioscience, San Jose, CA). Data were analyzed using BD FACSDiva software.
Concentrations	1 μM
Incubation time	2 h

Animal Experiment
Animal models	atherosclerotic lesions in Ldlr−/− mice
Formulation	saline
Dosages	10mg/kg
Administration	i.p.

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

Chemical Information

References

[1] Zhang XQ, et al. Adv Healthc Mater. Nanoparticles containing a liver X receptor agonist inhibit inflammation and atherosclerosis.

[2] Stukas S, et al. Biochim Biophys Acta. The LXR agonist GW3965 increases apoA-I protein levels in the central nervous system independent of ABCA1.

[3] Leik CE, et al. Br J Pharmacol. GW3965, a synthetic liver X receptor (LXR) agonist, reduces angiotensin II-mediated pressor responses in Sprague-Dawley rats.