FW1256 is a phenyl analog and a slow-release hydrogen sulfide (H2S) donor. FW1256 inhibits NF-κB activity, causes apoptosis, has effective anti~inflammatory effects, and may be utilized in study/research of cancer and cardiovascular diseases.

Physicochemical Properties

Molecular Formula	C12H10NOPS
Molecular Weight	247.253
Exact Mass	247.022
CAS #	117089-08-4
PubChem CID	13975035
Appearance	White to off-white solid powder
LogP	4.5
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	1
Heavy Atom Count	16
Complexity	303
Defined Atom Stereocenter Count	0
InChi Key	AXFLRXAQPCLTEA-UHFFFAOYSA-N
InChi Code	InChI=1S/C12H10NOPS/c16-15(10-6-2-1-3-7-10)13-11-8-4-5-9-12(11)14-15/h1-9H,(H,13,16)
Chemical Name	2-phenyl-2-sulfanylidene-3H-1,3,2λ5-benzoxazaphosphole
Synonyms	FW 1256; FW-1256; FW1256
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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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

ln Vitro	When LPS-stimulated RAW264.7 macrophages were treated with FW1256 (200 µM) for 24.5 hours, the levels of IL-1β, COX-2, and iNOS mRNA and protein were considerably reduced [1]. LPS-stimulated RAW264.7 macrophages showed a substantial reduction in IL-1β, COX-2, and iNOS PROTE as well as proteins after treatment with FW1256 (200 µM; 24.5 hours; AW264.7 cells) [1]. When applied to LPS-stimulated RAW264.7 macrophages and bone, FW1256 decreases TNF-α (IC50 of 61.2 µM), IL-6 (IC50 of 11.7 µM), PGE2 (IC50 of 25.5 µM), and NO (IC50 of 25.5 µM) in a concentration-dependent manner. Bone marrow-derived macrophages (BMDM) were produced at 34.6 µM (IC50 for TNF-α, IL-6, PGE2, and NO are 414.9 µM, 300.2 µM, 4 µM, and 9.5 µM, respectively) [1]. LPS-stimulated RAW264.7 macrophages treated with FW1256 exhibit decreased nuclear p65 levels and reduced cytosolic phospho-IκBα levels, both of which are indicators of decreased NF-κB activation [1].
ln Vivo	In LPS-treated mice, FW1256 treatment (100 mg/kg; i.p.; male C57BL/6 mice) decreased the levels of IL-1β, TNFα, nitrate/nitrite, and PGE2 [1].
Cell Assay	RT-PCR[1] Cell Types: RAW264.7 Cell Tested Concentrations: 200 µM Incubation Duration: 24.5 hrs (hours) Experimental Results: IL-1β, COX-2 and iNOS mRNA were Dramatically diminished in LPS-stimulated RAW264.7 macrophages. Western Blot Analysis [ 1 ] Cell Types: RAW264.7 Cell Tested Concentrations: 200 µM Incubation Duration: 24.5 hrs (hours) Experimental Results: IL-1β, COX-2 and iNOS proteins were Dramatically diminished in LPS-stimulated RAW264.7 macrophages.
Animal Protocol	Animal/Disease Models: Male C57BL/6 mice (20-25 g, 6-10 weeks) were injected with E. coli lipopolysaccharide (LPS) [1] Doses: 100 mg/kg Route of Administration: intraperitoneal (ip) injection Experimental Results: IL-1β, TNFα , Nitrate/nitrite and PGE2 levels in LPS-treated mice.
References	[1]. Huang CW, et al. A novel slow-releasing hydrogen sulfide donor, FW1256, exerts anti-inflammatory effects in mouse macrophages and in vivo. Pharmacol Res. 2016 Nov;113(Pt A):533-546. [2]. Feng W, et al. Discovery of New H2S Releasing Phosphordithioates and 2,3-Dihydro-2-phenyl-2-sulfanylenebenzo[d][1,3,2]oxazaphospholes with Improved Antiproliferative Activity. J Med Chem. 2015 Aug 27;58(16):6456-80.

Solubility Data

Solubility (In Vitro)

DMSO : ≥ 250 mg/mL (~1011.12 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	4.0445 mL	20.2224 mL	40.4449 mL
	5 mM	0.8089 mL	4.0445 mL	8.0890 mL
	10 mM	0.4044 mL	2.0222 mL	4.0445 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.