Copanlisib HCl (formerly BAY-806946; BAY 80-6946; Aliqopa), the hydrochloride salt of Copanlisib, is a pan-class I PI3K (phosphoinositide 3-kinase) inhibitor approved in 2017 for the treatment of relapsed follicular lymphoma. In cell-free assays, it inhibits PI3Kα/β/γ/δ with IC50 values of 0.5, 3.7, 6.4, and 0.7 nM, respectively. The FDA approved copanlisib in 2017 for the treatment of adults with relapsed follicular lymphoma. Phosphoinositide 3-kinase (PI3K) inhibitor BAY 80-6946 has potential anticancer properties and inhibits proliferation in HuCCT-1 (KRASG12D) and EGI-1 (KRASG12D) cell lines with IC50 values of 147 nM and 137 nM, respectively. The maximum tolerated dose (MTD) of BAY 80-6946 is generally well tolerated at 0.8 mg/kg. The results of pharmacokinetics (PK) support weekly dosing. In the first 24 hours following a MTD dose, hyperglycemia of grade 2 or 3 may occur. Clinical SD, pharmacokinetics, and FDG-PET data all support effective exposure and PI3K pathway inhibition.

Physicochemical Properties

Molecular Formula	C23H30CL2N8O4
Molecular Weight	553.45
Exact Mass	552.176
Elemental Analysis	C, 49.92; H, 5.46; Cl, 12.81; N, 20.25; O, 11.56
CAS #	1402152-13-9
Related CAS #	Copanlisib;1032568-63-0
PubChem CID	135565785
Appearance	Off-white to light yellow solid powder
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	9
Rotatable Bond Count	7
Heavy Atom Count	37
Complexity	974
Defined Atom Stereocenter Count	0
SMILES	Cl.Cl.O1CCN(CC1)CCCOC1C=CC2C(C=1OC)=N/C(=N\C(C1=CN=C(N)N=C1)=O)/N1CCNC1=2
InChi Key	STGQPVQAAFJJFX-UHFFFAOYSA-N
InChi Code	InChI=1S/C23H28N8O4.2ClH/c1-33-19-17(35-10-2-6-30-8-11-34-12-9-30)4-3-16-18(19)28-23(31-7-5-25-20(16)31)29-21(32)15-13-26-22(24)27-14-15;;/h3-4,13-14H,2,5-12H2,1H3,(H2,24,26,27)(H,28,29,32);2*1H
Chemical Name	2-amino-N-[7-methoxy-8-(3-morpholin-4-ylpropoxy)-2,3-dihydroimidazo[1,2-c]quinazolin-5-yl]pyrimidine-5-carboxamide dihydrochloride
Synonyms	BAY 80-6946; BAY80-6946; BAY-80-6946; BAY806946; BAY-806946; BAY 806946; Copanlisib HCl; Copanlisib dihydrochloride; Copanlisib; trade name Aliqopa
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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.
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	PI3Kα (IC50 = 0.5 nM); PI3Kδ (IC50 = 0.7 nM); PI3Kβ (IC50 = 3.7 nM); PI3Kγ (IC50 = 6.4 nM); mTOP (IC50 = 45 nM)
ln Vitro	In both KPL4 cells and LPA-stimulated PC3 cells, BAY 80-6946 reduces pAKT levels. BAY 80-6946 exhibits antiproliferative activity and induces apoptosis in a subset of human cancer cell lines that have PIK3CA mutations and/or overexpression of HER2. Combining HER2-targeted therapies with BAY 80-6946 inhibits growth more effectively than either therapy when used separately and can improve cells' sensitivity to trastuzumab and lapatinib.
ln Vivo	Copanlisib (BAY 80-6946; 0.5-6 mg/kg; intravenous injection; every second day, every third day; for 60 days; athymic nude rats) treatment exhibits significant antitumor activity in the rat KPL4 tumor xenograft model[1].
Enzyme Assay	The effect of BAY 80-6946 on PI3Kα, PI3Kβ, and PI3Kγ activity is measured by the inhibition of 33P incorporation into phosphatidylinositol (PI) in 384-well MaxiSorp® plates coated with 2 µg/well of PI and phosphatidylserine (PS) (1:1 molar ratio). In each PI3K isoform assay, 9 µL of reaction buffer (50 mM MOPSO, pH 7.0, 100 mM NaCl, 4 mM MgCl2, 0.1% BSA) is used, containing either 25 ng of purified human p110γ protein or 7.5 ng of His-tagged N-terminally truncated p110α or p110β protein. Adding 5 µL liters of a 40-µM ATP solution with 20 µCi/mL [33>/sup>P]-ATP initiates the reaction. By adding 5 µL of a 25-mM EDTA solution, the reaction is stopped after 2 hours of incubation at room temperature. After washing the plates, Ultima GoldTM scintillation cocktail (25 µL) is added. With the help of a BetaPlate Liquid Scintillation Counter, the radioactivity incorporated into the immobilized PI substrate is identified.
Cell Assay	The CellTiter-Glo® luminescent cell viability kit is used to calculate the rate of cell proliferation over a 72-hour period. In separate microtiter plates, cells are briefly plated. After an overnight incubation at 37 °C, the luminescence values in the t=0 hour plates are calculated. The cells are incubated in the t=72 hour plates for 72 hours at 37 °C after the addition of test substances that have been diluted in growth medium. After a 10-minute reaction with CellTiter-Glo® solution, Luminescence values are calculated using a Wallac 1420 Victor2TM 1420 multilabel HTS counter. Subtracting the luminescence values from the corresponding values in the t=72 hour plates, one can calculate the percentage inhibition of cell growth. Calculating the percentage of cell growth inhibition is done by comparing values between drug-treated cells and controls.
Animal Protocol	Athymic nude rats injected with KPL4 tumor cells[1] 0.5 mg/kg, 1 mg/kg, 3 mg/kg or 6 mg/kg Intravenous injection; every second day, every third day; for 60 days
References	[1]. BAY 80-6946 is a highly selective intravenous PI3K inhibitor with potent p110α and p110δ activities in tumor cell lines and xenograft models. Mol Cancer Ther. 2013 Nov;12(11):2319-30.
Additional Infomation	Copanlisib dihydrochloride is the dihydrochloride salt of copanlisib. It has a role as an antineoplastic agent, an apoptosis inducer and an EC 2.7.1.137 (phosphatidylinositol 3-kinase) inhibitor. It contains a copanlisib. See also: Copanlisib Dihydrochloride (annotation moved to).

Solubility Data

Solubility (In Vitro)	DMSO: ~10 mM Water: <1 mg/mL Ethanol: <1 mg/mL
Solubility (In Vivo)	Solubility in Formulation 1: 100 mg/mL (180.69 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication. Solubility in Formulation 2: 10% Trifluoroacetic acid water solution: 1mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.8068 mL	9.0342 mL	18.0685 mL
	5 mM	0.3614 mL	1.8068 mL	3.6137 mL
	10 mM	0.1807 mL	0.9034 mL	1.8068 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.