PD-L1-IN-2 inhibits PD-L1 and is a potential tumor immune agent. PD-L1-IN-2 is a Naamidine J analogue that exerts anti-tumor effects in vivo by reducing PD-L1 expression and enhancing tumor-infiltrating T cell immunity. PD-L1-IN-2 for colorectal cancer research.

Physicochemical Properties

Molecular Formula	C33H38N4O6
Molecular Weight	586.678028583527
Exact Mass	586.279
CAS #	2894733-91-4
PubChem CID	168355652
Appearance	White to off-white solid powder
LogP	5.6
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	12
Heavy Atom Count	43
Complexity	891
Defined Atom Stereocenter Count	0
InChi Key	WCBUHIGCYKSAAG-UHFFFAOYSA-N
InChi Code	InChI=1S/C33H38N4O6/c1-33(2,3)43-32(39)34-24-10-8-9-23(20-24)30(38)36-31-35-26(17-21-11-14-25(40-5)15-12-21)27(37(31)4)18-22-13-16-28(41-6)29(19-22)42-7/h8-16,19-20H,17-18H2,1-7H3,(H,34,39)(H,35,36,38)
Chemical Name	tert-butyl N-[3-[[5-[(3,4-dimethoxyphenyl)methyl]-4-[(4-methoxyphenyl)methyl]-1-methylimidazol-2-yl]carbamoyl]phenyl]carbamate
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	Compound 11c, PD-L1-IN-2, inhibits RKO cells with an IC50 value of 31.7μM[1]. In RKO cells, PD-L1 expression is inhibited by PD-L1-IN-2 (0-10 μM; 0-24 hours) in a way that is dependent on both dose and time [1]. PD-L1-IN-2 (0–10 μM; 0–24 hours) increases the turnover of the PD-L1 protein. The PD-L1 turnover rate in PD-L1-IN-2-treated cells was higher in CHX pulse chase compared to untreated cells[1].
ln Vivo	GPR84 antagonist 3 (compound 42) (ip; 25/50 mg/kg; once daily; 16 days) had a 45% inhibition rate in the 50 mg/kg group, suggesting that it can diminish tumor size. Compared to the PBS group, the group's average tumor weight was noticeably reduced[1].
Cell Assay	Western Blot Analysis[1] Cell Types: RKO cells Tested Concentrations: 5 μM Incubation Duration: 0h, 1h, 3h, 6h, 9h, 12h Experimental Results: Promoted the turnover of PD-L1 protein.
Animal Protocol	Animal/Disease Models: C57BL/6 mice with subcutaneous (sc) MC38 tumors[1] Doses: 25/50 mg/kg Route of Administration: ip; 25/50 mg/kg; one time/day; 16 days Experimental Results: Suppressed MC38 tumor growth in vivo.
References	[1]. Bioactivity-Driven Synthesis of the Marine Natural Product Naamidine J and Its Derivatives as Potential Tumor Immunological Agents by Inhibiting Programmed Death-Ligand 1. J Med Chem. 2023 Apr 27;66(8):5427-5438.

Solubility Data

Solubility (In Vitro)

DMSO: 50 mg/mL (85.23 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.26 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	1.7045 mL	8.5225 mL	17.0451 mL
	5 mM	0.3409 mL	1.7045 mL	3.4090 mL
	10 mM	0.1705 mL	0.8523 mL	1.7045 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.