BNTA is a potent extracellular matrix (ECM) modulator that promotes the synthesis of cartilage structural molecules on chondrocytes by activating superoxide dismutase 3 (SOD3). BNTA shows a promising potential for alleviating osteoarthritis by regulating chondrogenesis.

Physicochemical Properties

Molecular Formula	C17H11BRCLNO3S2
Molecular Weight	456.7611
Exact Mass	454.905
CAS #	685119-25-9
PubChem CID	2819453
Appearance	White to off-white solid powder
LogP	5.4
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	4
Heavy Atom Count	25
Complexity	577
Defined Atom Stereocenter Count	0
SMILES	BrC1=C(C(=C([H])S1)S(C1C([H])=C([H])C([H])=C([H])C=1[H])(=O)=O)N([H])C(C1=C([H])C([H])=C([H])C([H])=C1Cl)=O
InChi Key	OCNJYMSNHNAZON-UHFFFAOYSA-N
InChi Code	InChI=1S/C17H11BrClNO3S2/c18-16-15(20-17(21)12-8-4-5-9-13(12)19)14(10-24-16)25(22,23)11-6-2-1-3-7-11/h1-10H,(H,20,21)
Chemical Name	N-[4-(benzenesulfonyl)-2-bromothiophen-3-yl]-2-chlorobenzamide
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

ln Vitro	The cell viability of rat primary chondrocytes and human osteoarthritis chondrocytes is not affected by BNTA (0.01-10 μM; 1-7 d)[1]. SOX9 protein is significantly increased by BNTA (0.1 μM; 2 d)[1]. BNTA (0.1 μM; 2 d) significantly raises the levels of SOX9 and COL2A1 proteins in rat OA chondrocytes generated by IL1β[1]. In ATDC5 cells, BNTA (10 μM; 5 d) enhances proteoglycan staining[1]. In human OA chondrocytes, BNTA (0.01-10 μM; 6 h) increases the expression levels of ECM-related genes COL2A1, ACAN, proteoglycan 4 (PRG4), and SRY-box 9 (SOX9)[1]. In IL1β-induced rat OA chondrocytes, BNTA (0.01-10 μM; 6 h) raises Col2a1, Acan, Prg4, and Sox9 mRNA levels; the effects peak at 0.1 μM[1]. In arthritic cartilage explants, BNTA (0.01-1 μM; 2 or 3 w) increases anabolism and reduces inflammatory response[1].
ln Vivo	In rats undergoing anterior cruciate ligament transection (ACLT), BNTA (0.015–1.5 mg/kg; intra-articular injection; twice weekly for 4 and 8 weeks) was able to slow the progression of osteoarthritis (OA)[1].
Cell Assay	Cell Viability Assay[1] Cell Types: Human OA chondrocytes Tested Concentrations: 0.01, 0.1, 1, 10 μM Incubation Duration: 1, 3, 5, 7 d Experimental Results: No toxicity was observed. Western Blot Analysis[1] Cell Types: Human OA chondrocytes Tested Concentrations: 0.1 μM Incubation Duration: 2 d Experimental Results: Elevated SOX9 protein compared with vehicle.
Animal Protocol	Animal/Disease Models: Male SD rats weighing 80 g are induced by ACLT[1] Doses: 0.015, 0.15, 1.5 mg/kg Route of Administration: Intra-articular injection; twice a week for 4 and 8 weeks Experimental Results: Attenuated post-traumatic osteoarthritis development after intra-articular injection for 4 and 8 weeks and was well tolerated.
References	[1]. A small molecule promotes cartilage extracellular matrix generation and inhibits osteoarthritis development. Nat Commun. 2019 Apr 23; 10(1): 1914.

Solubility Data

Solubility (In Vitro)

DMSO: 100 mg/mL (218.93 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.47 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.47 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1893 mL	10.9467 mL	21.8933 mL
	5 mM	0.4379 mL	2.1893 mL	4.3787 mL
	10 mM	0.2189 mL	1.0947 mL	2.1893 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.