FD223 is a potent and specific inhibitor of phosphoinositide 3-kinase delta (PI3Kδ). FD223 shows high potency (IC50=1 nM) and good selectivity towards other isomers (IC50s for α, β and γ are 51 nM, 29 nM and 37 nM respectively). FD223 effectively suppresses the proliferation/growth of acute myeloid leukemia (AML) cell lines by inhibiting p-AKT Ser473, resulting in G1 phase arrest of the cell cycle. FD223 has potential in the research of leukemias such as AML.

Physicochemical Properties

Molecular Formula	C17H12CLN5O2S
Molecular Weight	385.827480316162
Exact Mass	385.04
CAS #	2050524-24-6
PubChem CID	141461656
Appearance	Off-white to brown solid powder
LogP	2.9
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	4
Heavy Atom Count	26
Complexity	580
Defined Atom Stereocenter Count	0
SMILES	C1(S(NC2=CC(C3=CN=C4NN=CC4=C3)=CN=C2Cl)(=O)=O)=CC=CC=C1
InChi Key	VFNUYVJFDQOTNJ-UHFFFAOYSA-N
InChi Code	InChI=1S/C17H12ClN5O2S/c18-16-15(23-26(24,25)14-4-2-1-3-5-14)7-12(8-19-16)11-6-13-10-21-22-17(13)20-9-11/h1-10,23H,(H,20,21,22)
Chemical Name	N-[2-chloro-5-(1H-pyrazolo[3,4-b]pyridin-5-yl)pyridin-3-yl]benzenesulfonamide
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	PI3Kδ 1 nM (IC50) PI3Kα 51 nM (IC50) PI3Kβ 29 nM (IC50) PI3Kγ 37 nM (IC50)
ln Vitro	FD223 demonstrates significant anti-proliferative effects on the p110δ-positive AML cell lines KG-1, MOLM-16, HL-60, and EOL-1, with IC50 values of 2.82 μM, 5.82 μM, 0.87 μM, and 2.25 μM, in that order. With an IC50 value of 23.13 μM, FD223 exhibits weak anti-proliferative activity against the p110δ unexpressed MM.1R cell line[1]. The phosphorylation of Akt (Ser473) is dose-dependently reduced by FD223 (MOLM-16 cells; 0.1-5 μM; 16 hours), which is consistent with the positive control Idelalisib and shows that the PI3K/Akt pathway is blocked in MOLM-16 cells[1]. Like its positive control Idelalisib, FD223 (MOLM-16 cells; 24 hours; 1–5 μM) also stops the cell cycle at the G1 phase[1]. Cellular apoptosis is dose-dependently induced by FD223 (1–5 μM; 48 hours)[1].
ln Vivo	In the MOLM-16 xenograft model, FD223 (20 and 40 mg/kg; po, per day for 14 consecutive days) exhibits strong anticancer effectiveness with a 49% tumor volume decrease at a dose of 40 mg/kg/day (po). The preliminary safety assessment of the drug indicates no appreciable toxicity[1]. Following intravenous administration, FD223 (iv; dose of 2 mg/kg; po; 10 mg/kg rats) has a moderate plasma clearance rate (C = 0.191 L·h–1·kg–1). It has good oral plasma exposures (AUC0-∞>9000 h·ng/mL), an adequate oral bioavailability (17.6%), a half-life (t1/2) of 3.74 h, and a Cmax of 1104 ng/mL in the po route[1].
Cell Assay	Apoptosis Analysis[1] Cell Types: MOLM-16 cells Tested Concentrations: 1-5 μM Incubation Duration: 48 hrs (hours) Experimental Results: Dose-dependently induced cellular apoptosis, which is superior to that of positive control Idelalisib. Western Blot Analysis[1] Cell Types: MOLM-16 cells Tested Concentrations: 0.1-5 μM Incubation Duration: 16 hrs (hours) Experimental Results: Dose-dependently decreased phosphorylation of Akt (Ser473).
Animal Protocol	Animal/Disease Models: MOLM-16 xenograft model of BALB/c nude mice[1] Doses: 20 and 40 mg/kg Route of Administration: Po, per day for 14 days Experimental Results: demonstrated a dose-dependent tumor growth inhibition (TGI) of 31% for 20 mg/kg and 49% for 40 mg/kg
References	[1]. Bioisosteric replacements of the indole moiety for the development of a potent and selective PI3Kδ inhibitor: Design, synthesis and biological evaluation [published online ahead of print, 2021 Jun 21]. Eur J Med Chem. 2021;223:113661.

Solubility Data

Solubility (In Vitro)

DMSO : 100 mg/mL (259.18 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.48 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 (6.48 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.5918 mL	12.9591 mL	25.9182 mL
	5 mM	0.5184 mL	2.5918 mL	5.1836 mL
	10 mM	0.2592 mL	1.2959 mL	2.5918 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.