HEC96719 is a selective and orally bioactive tricyclic farnesoid X receptor (FXR) agonist/activator with EC50s of 1.37 and 1.55 nM via time-resolved fluorescence energy transfer (TR-FRET) and luciferase, respectively. Report the test. HEC96719 significantly improves non-alcoholic steatohepatitis (NASH) and liver fibrosis with good tissue distribution in the liver and intestines. HEC96719 may be utilized in the study of non-alcoholic steatohepatitis.

Physicochemical Properties

Molecular Formula	C29H22CL2N2O5
Molecular Weight	549.401385784149
Exact Mass	548.09
CAS #	2181834-03-5
PubChem CID	138675487
Appearance	White to off-white solid powder
LogP	6.3
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	38
Complexity	877
Defined Atom Stereocenter Count	0
InChi Key	JZJOXLSYULKNLW-UHFFFAOYSA-N
InChi Code	InChI=1S/C29H22Cl2N2O5/c30-19-2-1-3-20(31)24(19)25-18(26(38-33-25)15-4-5-15)14-36-23-9-8-21-27(32-23)29(10-11-29)13-17-7-6-16(28(34)35)12-22(17)37-21/h1-3,6-9,12,15H,4-5,10-11,13-14H2,(H,34,35)
Chemical Name	3-[[5-cyclopropyl-3-(2,6-dichlorophenyl)-1,2-oxazol-4-yl]methoxy]spiro[6H-[1]benzoxepino[3,2-b]pyridine-5,1'-cyclopropane]-9-carboxylic acid
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	EC50: 1.37 nM (FXR, TR-FRET), 1.55 nM (FXR, luciferase reporter assay)[1].
ln Vivo	It has been demonstrated that HEC96719 (0.5, 1.5, and 5 mg/kg; orally, once daily for 14 days) activates FXR via raising levels of fibroblast growth factor 15 (FGF15)[1]. FGF15 levels in the ileum and hepatic bile salt export pump (BSEP) levels are elevated by HEC96719 (5 mg/kg; oral, 1 dosage)[1]. In non-alcoholic steatohepatitis (NASH), HEC96719 (0.1, 0.3, and 1 mg/kg; orally, once day for 6 weeks) dramatically reduces symptoms[1]. HEC96719 (0.1, 0.3, and 1 mg/kg; taken orally once daily for four weeks) is better than obeticholic acid (OCA) in terms of improving liver fibrosis[1].
Animal Protocol	Animal/Disease Models: Male ob/ob nonalcoholic steatohepatitis (NASH) mouse models[1] Doses: 0.1, 0.3 and 1 mg/kg Route of Administration: Oral administration; 0.1, 0.3 and 1 mg/kg, one time/day for 6 weeks Experimental Results: diminished levels of serum alanine aminotransferase (ALT) and liver triglyceride (TG), dose-dependently increased NASH activity and decreased NASH activity score. Animal/Disease Models: Male C57BL/6 liver fibrosis mouse models[1] Doses: 0.1, 0.3 and 1 mg/kg Route of Administration: Oral administration; 0.1, 0.3 and 1 mg/kg, one time/day for 4 weeks Experimental Results: diminished levels of serum ALT and TBIL, and decreased fibrosis area.
References	[1]. Discovery of a tricyclic farnesoid X receptor agonist HEC96719, a clinical candidate for treatment of non-alcoholic steatohepatitis. Eur J Med Chem. 2022 Feb 15;230:114089.

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	1.8202 mL	9.1008 mL	18.2017 mL
	5 mM	0.3640 mL	1.8202 mL	3.6403 mL
	10 mM	0.1820 mL	0.9101 mL	1.8202 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.