JY-2 is a moderately selective and orally bioactive inhibitor of forkhead transcription factor O1 (FoxO1) with IC50 of 22 μM for inhibiting FoxO1 transcriptional activity. JY-2 has a moderate inhibitory effect on FoxO3a and FoxO4. JY-2 has antidiabetic activity.

Physicochemical Properties

Molecular Formula	C13H7CL2N3O
Molecular Weight	292.12
Exact Mass	290.996
CAS #	339103-05-8
PubChem CID	702923
Appearance	White to off-white solid powder
Density	1.4±0.1 g/cm3
Boiling Point	457.4±55.0 °C at 760 mmHg
Flash Point	230.4±31.5 °C
Vapour Pressure	0.0±1.1 mmHg at 25°C
Index of Refraction	1.613
LogP	4.02
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	2
Heavy Atom Count	19
Complexity	308
Defined Atom Stereocenter Count	0
InChi Key	WBMHQMQALJDGHI-UHFFFAOYSA-N
InChi Code	InChI=1S/C13H7Cl2N3O/c14-8-4-5-9(10(15)7-8)13-17-12(18-19-13)11-3-1-2-6-16-11/h1-7H
Chemical Name	5-(2,4-dichlorophenyl)-3-pyridin-2-yl-1,2,4-oxadiazole
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	22 μM (FoxO1 transcriptional activity)[1]
ln Vitro	In HepG2 and INS-1 cells, JY-2 (10-100 μM; 24 h) lessens the lipotoxicity caused by palmitic acid (PA; HY-N0830) [1].
ln Vivo	Mice treated with JY-2 (50-200 mg/kg; oral; three times for two days or daily for four weeks) do not exhibit diabetic effects[1]. The JY-2 pharmacokinetic parameters are as follows: parameters iv (20 mg/kg) po (50 mg/kg) AUCall (ng·h/mL) 5017 ± 1038 12270 ± 2775 AUCinf.obs (ng·h/mL) 5030 ± 1037 12400 ± 2753 Cmax (ng/mL) 10790 ± 3269 6826 ± 2342 Tmax (h) 0.1 ± 0.1 0.8 ± 0.7 T1/2 (h) 0.8 ± 0.2 1.3 ± 0.4 MRTinf.obs (h) 0.7 ± 0.1 2.0 ± 0. 1 F (%) 97.8
Cell Assay	Real Time qPCR[1] Cell Types: HepG2 and INS-1 cells Tested Concentrations: 10, 50 and 100 μM Incubation Duration: 24 h Experimental Results: diminished palmitic acid (PA)-induced G6Pase and PEPCK mRNA expression. Inhibited PA-induced lipid accumulation . diminished PA-induced mRNA expression of ER stress markers (ATF3, CHOP and GRP78). Western Blot Analysis[1] Cell Types: HepG2 cells Tested Concentrations: 10, 50 and 100 μM Incubation Duration: 4 h; in the presence of PA ( 500μM) Experimental Results: Increased p-FoxO1 levels in the whole cell lysate with a concurrent reduction in nuclear FoxO1 levels.
Animal Protocol	Animal/Disease Models: C57BL/6J mice[1] Doses: 50, 100, 200 mg/kg Route of Administration: po (oral gavage) three times for two days (9:00 AM, 7:00 PM, 9:00 AM on the next day) Experimental Results: Improved glucose tolerance. Dramatically decreased the expression of G6Pase and PEPCK mRNA in the liver. Enhanced mRNA expression of insulin and PDX-1 in the pancreas. Animal/Disease Models: db/db mice and C57BL/6J mice, high fat-diet-induced obese diabetic (DIO) model[1] Doses: 50, 100 mg/kg Route of Administration: po (oral gavage) one time/day for 4 weeks Experimental Results: diminished the levels of fasting blood glucose, improved glucose tolerance. The expression of ColIV, a fibrosis marker, was also lowered. Animal/Disease Models: C57BL/6J mice[1] Doses: 20 mg/kg or 50 mg/kg Route of Administration: IV or PO (pharmacokinetic/PK Analysis) Experimental Results: demonstrated an overall good pharmacokinetic/PK profile.
References	[1]. Novel FoxO1 inhibitor, JY-2, ameliorates palmitic acid-induced lipotoxicity and gluconeogenesis in a murine model. Eur J Pharmacol. 2021 May 15;899:174011.

Solubility Data

Solubility (In Vitro)

DMSO: 125 mg/mL (427.91 mM)

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	3.4233 mL	17.1163 mL	34.2325 mL
	5 mM	0.6847 mL	3.4233 mL	6.8465 mL
	10 mM	0.3423 mL	1.7116 mL	3.4233 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.