F44-A13 is an orally active, highly selective farnesoid X receptor (FXR) antagonist with an IC50 of 1.1 μM. F44-A13 can optimize cholesterol metabolism and reduce the activity of CYP7A1 by inducing its expression. F44-A13 can reduce the levels of cholesterol, triglycerides, and low-density lipoprotein cholesterol (LDL-C) in mouse models. F44-A13 can be used to study metabolic diseases associated with lipid disorders.

Physicochemical Properties

Molecular Formula	C28H40N4O5S
Molecular Weight	544.71
CAS #	1338190-14-9
Appearance	Yellow to brown solid powder
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

Targets	PPARα PPARβ RXR α RXRβ RXRγ RORγ
ln Vitro	F44-A13 is highly selective for multiple nuclear receptors, including retinoic acid receptor α/β/γ (RARα/β/γ), retinoid X receptor α/β/γ, (RXRα/β/γ), pregnane X receptor (PXR), peroxisome proliferator-activated receptor α/β (PPARα/β), thyroid hormone receptor β (THRβ), and retinoic acid receptor-related orphan receptor γ (RORγ)[1]. F44-A13 (50 μM, 24 hours) inhibited the transcriptional activity of FXR in a dose-dependent manner in the presence of 50 μM CDCA (HY-76847) with an IC50 value of 3.0 μM. F44-A13 is a low-toxic, highly selective FXR antagonist[1]. F44-A13 (F44-A13: 3, 10, 30 μM; CDCA: 100 μM; 24 hours) can improve cholesterol metabolism and reduce cholesterol activity by inducing the expression of CYP7A1. F44-A13 can reduce the expression levels of Shp and Bsep and increase the expression level of CYP7A1[1].
ln Vivo	F44-A13 (20, 40 mg/kg; oral and intraperitoneal injection; 4 days) effectively reduced total cholesterol (TC), triglycerides (TG) and low-density lipoprotein cholesterol in C57BL/6 mice (LDL-C) levels[1].
Cell Assay	Cell Viability Assay[1] Cell Types: HepG2 , HEK293A Tested Concentrations: 100 μM Incubation Duration: 24h Experimental Results: Was not cytotoxic to HepG2 and HEK293A cells RT-PCR[1] Cell Types: HepG2 Tested Concentrations: F44-A13: 3, 10, 30 μM; CDCA: 100 μM Incubation Duration: 24h Experimental Results: Was able to reverse the regulation of FXR downstream target genes Shp, Bsep and Cyp7a1 by CDCA in a dose-dependent manner. Decreased the expression level of Shp and Bsep, and increased the expression level of Cyp7a1.
Animal Protocol	Animal/Disease Models:C57BL/6 mice[1] Doses: 20, 40 mg/kg Route of Administration: Intraperitoneal injection (i.p.) and Oral gavage (p.o.); 4 days Experimental Results: Intraperitoneal injection significantly reduced TC levels by more than 28%, and oral administration significantly reduced TC levels in a dose-dependent manner. Reduced TG levels by more than 30% at both 20 and 40 mg/kg orally, while intraperitoneal injection did not significantly reduce TG levels. Oral doses of 20 and 40 mg/kg were effective in reducing LDL-C levels by 12% and 23%, and intraperitoneal injections by 38%.
References	[1]. Discovery of novel and selective farnesoid X receptor antagonists through structure-based virtual screening, preliminary structure-activity relationship study, and biological evaluation. Eur J Med Chem. 2024;269:116323.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.8358 mL	9.1792 mL	18.3584 mL
	5 mM	0.3672 mL	1.8358 mL	3.6717 mL
	10 mM	0.1836 mL	0.9179 mL	1.8358 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.