WM-3835 (WM3835) is a novel and potent lysine acetyltransferase HBO1 (KAT7) inhibitor with potential anticancer activity. It binds directly to the acetyl-CoA binding site of HBO1 33. HBO1 (KAT7 or MYST2) is a histone acetyltransferase that acetylates H3 and H4 histones. HBO1 overexpression promotes OS cell growth in vitro and in vivo. WM-3835 was able to potently suppressed OS cell proliferation and migration, and led to apoptosis activation. Furthermore, intraperitoneal injection of a single dose of WM-3835 potently inhibited OS xenograft growth in SCID mice.

Physicochemical Properties

Molecular Formula	C20H17FN2O4S
Molecular Weight	400.4234
Exact Mass	400.09
Elemental Analysis	C, 59.99; H, 4.28; F, 4.74; N, 7.00; O, 15.98; S, 8.01
CAS #	2229025-70-9
PubChem CID	134581412
Appearance	White to off-white solid powder
LogP	3.8
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	5
Heavy Atom Count	28
Complexity	633
Defined Atom Stereocenter Count	0
InChi Key	KVJFJJXCBRSCDY-UHFFFAOYSA-N
InChi Code	InChI=1S/C20H17FN2O4S/c1-13-10-15(14-6-3-2-4-7-14)11-18(19(13)21)20(25)22-23-28(26,27)17-9-5-8-16(24)12-17/h2-12,23-24H,1H3,(H,22,25)
Chemical Name	2-fluoro-N'-(3-hydroxyphenyl)sulfonyl-3-methyl-5-phenylbenzohydrazide
Synonyms	WM 3835; WM3835; WM-3835;
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: This product requires protection from light (avoid light exposure) during transportation and storage.
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	pOS-1 cell viability is inhibited by WM-3835 (1-25 uM; 24-96 hours) in a concentration-dependent manner [1]. In pOS-1 cells, WM-3835 (5 uM; 72 h) dramatically raises the proportion of TUNEL-positive nuclei and initiates apoptosis [1]. In pOS-1 cells, WM-3835 (5 μM; 24 hours) reduces the expression of MYLK-HOXA9 mRNA [1]. Inhibiting H4K12ac-H3K14ac in a dose-dependent manner is WM-3835 (1-25 uM). Total H3 and H4 histones as well as the expression of the HBO1 protein are not altered by WM-3835 [1]. When it comes to HBO1-KO pOS-1 cells, koHBO1-1 and koHBO1-2, and HBO1-low human osteoblasts, WM-3835 (5 μM) cannot cause apoptosis or decreased viability [1].
ln Vivo	The growth of pOS-1 xenografts in SCID mice is effectively inhibited by WM-3835 (10 mg/kg/day; i.p.; for 21 days) [1].
Cell Assay	Cell viability assay [1] Cell Types: primary human OS (pOS-1) Cell Tested Concentrations: 1, 5, 10, 25 uM Incubation Duration: 24, 48, 72, 96 hrs (hours) Experimental Results: A certain concentration inhibits pOS-1 cells Vitality-dependent manner. Exhibits significant anti-survival activity for at least 48 hrs (hours), showing time dependence. Apoptosis analysis[1] Cell Types: pOS-1 Cell Tested Concentrations: 5 uM Incubation Duration: 72 hrs (hours) Experimental Results: Activation of apoptosis and significant increase in TUNEL-positive nuclei. RT-PCR[1] Cell Types: pOS-1 Cell Tested Concentrations: 5 uM Incubation Duration: 24 hrs (hours) Experimental Results: MYLK-HOXA9 mRNA expression was downregulated.
Animal Protocol	Animal/Disease Models: SCID (severe combined immunodeficient) mouse (18-19 g, female) [1] with pOS1 cells Doses: 10 mg/kg Route of Administration: IP; daily; continued for 21 days Experimental Results: Effective inhibition of pOS-1 xenografts grow.
References	[1]. The histone acetyltransferase HBO1 functions as a novel oncogenic gene in osteosarcoma. Theranostics. 2021 Mar 4;11(10):4599-4615.

Solubility Data

Solubility (In Vitro)

DMSO : ~250 mg/mL (~624.34 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.19 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 20.8 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: ≥ 2.08 mg/mL (5.19 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (5.19 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 20.8 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	2.4974 mL	12.4869 mL	24.9738 mL
	5 mM	0.4995 mL	2.4974 mL	4.9948 mL
	10 mM	0.2497 mL	1.2487 mL	2.4974 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.