RBPJ Inhibitor-1 (RIN1) is the first RBPJ inhibitor that blocks the interaction between RBPJ and SHARP. RBPJ Inhibitor-1 (RIN1) inhibits Notch-dependent tumor cell proliferation/growth.

Physicochemical Properties

Molecular Formula	C17H14FN3O2
Molecular Weight	311.310367107391
Exact Mass	311.107
CAS #	2682114-39-0
PubChem CID	146019721
Appearance	Light yellow to light brown solid powder
LogP	2.4
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	4
Heavy Atom Count	23
Complexity	423
Defined Atom Stereocenter Count	0
SMILES	C(N)(=O)C1=CC=C(C2N(C)N=CC=2)C=C1OC1=CC=CC=C1F
InChi Key	MFSSHRCJKRDIOL-UHFFFAOYSA-N
InChi Code	InChI=1S/C17H14FN3O2/c1-21-14(8-9-20-21)11-6-7-12(17(19)22)16(10-11)23-15-5-3-2-4-13(15)18/h2-10H,1H3,(H2,19,22)
Chemical Name	2-(2-fluorophenoxy)-4-(2-methylpyrazol-3-yl)benzamide
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	RIN1 suppresses the growth of blood cancer cell lines and stimulates skeletal muscle development from C2C12 myoblasts [1]. RIN1 (0.6 μM, equivalent to 3 times IC50) has been shown to cause the production of multinucleated myofibers because it decreases the number of MHC+ cells and increases the number of nuclei per cell [1].
Cell Assay	Cell Viability Assay[1] Cell Types: AD-293 cells were transfected with RBPJ-VP16myc. Tested Concentrations: 2 μM. Incubation Duration: 17 hrs (hours). Experimental Results: Inhibited Hes1-Luciferase activity with an IC50 of 0.18 µM and Emax of 82%. Inhibited NOTCH3 ICD with similar potency and efcacy (0.19 µM and Emax=88%). Cell Proliferation Assay[1] Cell Types: T-cell acute lymphoblastic leukemia (T-ALL) patients (Jurkat and KOPT-K1) and in the mantle cell lymphoma (MCL) line REC-1. Tested Concentrations: 0.1-10 μM. Incubation Duration: 96 h. Experimental Results: Inhibited NOTCH-dependent tumor cell proliferation. Was active in both the cancer cell anti-proliferation and myoblast diferentiation assays.
References	[1]. Disruption of NOTCH signaling by a small molecule inhibitor of the transcription factor RBPJ. Sci Rep. 2019 Jul 25;9(1):10811.

Solubility Data

Solubility (In Vitro)

DMSO: 250 mg/mL (803.06 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (6.68 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 (6.68 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 (6.68 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	3.2122 mL	16.0612 mL	32.1223 mL
	5 mM	0.6424 mL	3.2122 mL	6.4245 mL
	10 mM	0.3212 mL	1.6061 mL	3.2122 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.