PD-1/PD-L1-IN-9 is a potent and orally bioactive PD-1/PD-L1 interaction inhibitor (antagonist) with IC50 of 3.8 nM. PD-1/PD-L1-IN-9 can enhance the killing activity of immune cells against tumor cells. PD-1/PD-L1-IN-9 exhibited significant in vivo anticancer effect in the CT26 mouse model.

Physicochemical Properties

Molecular Formula	C22H24N2O2
Molecular Weight	348.438165664673
Exact Mass	348.183
CAS #	2628506-54-5
Related CAS #	PD-1/PD-L1-IN-9 hydrochloride
PubChem CID	155925913
Appearance	White to off-white solid powder
LogP	3.5
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	7
Heavy Atom Count	26
Complexity	400
Defined Atom Stereocenter Count	0
SMILES	O(C)C1=C(C=CC(C2C=CC=C(C3C=CC=CC=3)C=2C)=N1)CNCCO
InChi Key	ANLIWBJMRQBWBM-UHFFFAOYSA-N
InChi Code	InChI=1S/C22H24N2O2/c1-16-19(17-7-4-3-5-8-17)9-6-10-20(16)21-12-11-18(15-23-13-14-25)22(24-21)26-2/h3-12,23,25H,13-15H2,1-2H3
Chemical Name	2-[[2-methoxy-6-(2-methyl-3-phenylphenyl)pyridin-3-yl]methylamino]ethanol
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

Targets	IC50: 3.8 nM (PD-1/PD-L1)[1]
ln Vitro	Compound 24 (PD-1/PD-L1-IN-9; 46.9-1500 nM; 2 h pretreatment) dose-dependently stimulates the anti-tumor immunity of peripheral blood mononuclear cells (PBMCs) against MDB-MB 231 cells; the EC50 is roughly 100 nM[1].
ln Vivo	PD-1/PD-L1-IN-9 (Compound 24; 40-80 mg/kg; PO; once daily for 2 weeks) does not result in weight loss or death[1], but it does limit tumor growth in a dose-dependent manner. PD-1/PD-L1-IN-9 (3 mg/kg; intravenous injection; single dosage) had a half-life of 4.2 hours, a plasma clearance of 11.5 L/h/kg, and a Cmax of 1233 ng/mL in rats[1]. Rats with Cmax = 192 ng/mL show moderate oral bioavailability (F = 22%) and half-life (t1/2 = 6.4 h) for PD-1/PD-L1-IN-9 (25 mg/kg; oral; single dose)[1].
Animal Protocol	Animal/Disease Models: Male balb/c (Bagg ALBino) mouse (5-6 weeks) were inoculated CT26 cells[1] Doses: 40 mg/kg, 80 mg/kg Route of Administration: Oral gavage; one time/day, for 2 weeks Experimental Results: Dramatically diminished the final tumor weight, with TGI values of 60 and 67% at the dose of 40 and 80 mg/kg, respectively. Animal/Disease Models: pharmacokinetic/PK analysis in SD (Sprague-Dawley) rats[1] Doses: 3 mg/kg and 25 mg/kg Route of Administration: intravenous (iv) injection or oral gavage; single dose Experimental Results: Route Dose (mg/kg) AUC(0-t) (ng·h/mL) Cmax (ng/mL) t1/2 (h) Tmax Cl (L·h /kg) Vz (L/kg) F (%) iv 3 430.5 1233 4.2 0.03 11.5 78.6 / po 25 787.4 192 6.4 0.69 28.8 249.3 22
References	[1]. Novel Biphenyl Pyridines as Potent Small-Molecule Inhibitors Targeting the Programmed Cell Death-1/Programmed Cell Death-Ligand 1 Interaction. J Med Chem. 2021 May 30.

Solubility Data

Solubility (In Vitro)

DMSO: 100 mg/mL (286.99 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.17 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 (7.17 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 25.0 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: ≥ 2.5 mg/mL (7.17 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	2.8699 mL	14.3497 mL	28.6993 mL
	5 mM	0.5740 mL	2.8699 mL	5.7399 mL
	10 mM	0.2870 mL	1.4350 mL	2.8699 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.