J22352 is a selective HDAC6 inhibitor (antagonist) with similar targeted chimeric proteolysis (PROTAC) properties, with IC50 of 4.7 nM. J22352 promotes HDAC6 by inhibiting glioblastoma autophagy and inducing anti-tumor immune responses. Degrade and induce anti-cancer effects, and restore host anti-tumor activity by reducing the immunosuppressive activity of PD-L1.

Physicochemical Properties

Molecular Formula	C24H27N3O4
Molecular Weight	421.488886117935
Exact Mass	415.153
CAS #	2252395-44-9
PubChem CID	138377601
Appearance	Light yellow to yellow solid powder
LogP	3
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	6
Heavy Atom Count	31
Complexity	655
Defined Atom Stereocenter Count	0
SMILES	O=C1C2CCCCC2N(CC2C=CC(C(NO)=O)=CC=2)C(N1CCC1C=CC=CC=1)=O
InChi Key	JBJIKUXUKSADFV-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H21N3O4/c28-22(25-31)19-12-10-18(11-13-19)16-27-21-9-5-4-8-20(21)23(29)26(24(27)30)15-14-17-6-2-1-3-7-17/h1-13,31H,14-16H2,(H,25,28)
Chemical Name	4-[[2,4-dioxo-3-(2-phenylethyl)quinazolin-1-yl]methyl]-N-hydroxybenzamide
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	J22352 (0.1–20 μM; 72 hours) dose-dependently decreases the viability of U87MG cells[1]. J22352 (10 μM; 24 hours) decreases the quantity of HDAC6 protein in a dose-dependent manner [1].
ln Vivo	J22352 (10 mg/kg; intraperitoneally injected daily for 14 days in nude hamster mice) resulted in a >80% tumor growth inhibition (TGI) rate. J22352 is well tolerated in mice [1].
Cell Assay	Cell Viability Assay[1] Cell Cell Types: U87MG Cell Tested Concentrations: 0.1 μM; 0.5 μM; 1 μM; 2.5 μM; 5 μM; 10 μM; 20 μM Incubation Duration: 72 hrs (hours) Experimental Results: U87MG cell proliferation diminished in a dose-dependent manner. Western Blot Analysis[1] Cell Types: U87MG Cell Tested Concentrations: 10 μM Incubation Duration: 24 hrs (hours) Experimental Results: Aberrant overexpression of HDAC6 in glioblastoma demonstrated a dose-dependent decrease.
Animal Protocol	Animal/Disease Models: Male nude mice (BALB/cAnN.Cg-Foxnlnu/CrlNarl, 4-6 weeks old) [1] Doses: 10 mg/kg Route of Administration: intraperitoneal (ip) injection; one time/day for 14 days Experimental Results: Significant Anti-tumor effects and well tolerated in mice.
References	[1]. High-selective HDAC6 inhibitor promotes HDAC6 degradation following autophagy modulation and enhanced antitumor immunity in glioblastoma. Biochem Pharmacol. 2019 May; 163:458-471.

Solubility Data

Solubility (In Vitro)

DMSO : ~125 mg/mL (~300.89 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.01 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 (5.01 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 (5.01 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	2.3725 mL	11.8627 mL	23.7254 mL
	5 mM	0.4745 mL	2.3725 mL	4.7451 mL
	10 mM	0.2373 mL	1.1863 mL	2.3725 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.