UPF 1069 (UPF-1069), an isoquinolinone derivative, is a novel, potent and selective inhibitor of poly-(ADP-ribose) polymerase 2 (PARP-2) with potential neurprotective and anti-ischemic effects in vivo. It inhibits PARP-2, with IC50 values for PARP-1 and PARP-2 of 8 and 0.3 μM, respectively. It can decrease PAR formation and cause apoptosis in nuclear extracts from PARP-1-/-fibroblasts as well as in recombinant enzyme preparations. With an IC50 of 8 μmol/L, UPF 1069 exhibits greater selectivity for PARP-2 compared to PARP-1. It has been applied to study the function of PARP-1 and PARP-2 in brain damage following an ischemia.

Physicochemical Properties

Molecular Formula	C17H13NO3
Molecular Weight	279.29
Exact Mass	279.089
Elemental Analysis	C, 73.11; H, 4.69; N, 5.02; O, 17.19
CAS #	1048371-03-4
Related CAS #	1048371-03-4
PubChem CID	25015515
Appearance	White to off-white solid powder
Density	1.3±0.1 g/cm3
Boiling Point	570.3±50.0 °C at 760 mmHg
Melting Point	166-168 °C
Flash Point	298.7±30.1 °C
Vapour Pressure	0.0±1.6 mmHg at 25°C
Index of Refraction	1.618
LogP	2.22
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	3
Rotatable Bond Count	4
Heavy Atom Count	21
Complexity	426
Defined Atom Stereocenter Count	0
InChi Key	JJWMRRNGWSITSQ-UHFFFAOYSA-N
InChi Code	InChI=1S/C17H13NO3/c19-15(12-5-2-1-3-6-12)11-21-16-8-4-7-14-13(16)9-10-18-17(14)20/h1-10H,11H2,(H,18,20)
Chemical Name	5-phenacyloxy-2H-isoquinolin-1-one
Synonyms	UPF 1069; UPF1069; UPF-1069
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	PARP-2 ( IC50 = 0.3 μM ); PARP-1 ( IC50 = 8 μM )
ln Vitro	PF 1069 (Compound 55) is a PARP inhibitor; its IC50 values for PARP-1 and PARP-2 are 8 and 0.3 μM, respectively[1]. While UPF 1069 (1 µM) only marginally inhibits the enzymatic activity in wild-type fibroblasts, it reduces the residual PARP activity in PARP-1-deficient fibroblasts by about 80%. 0.1–1 µM UPF 1069 significantly increases CA1 hippocampal damage. Oxygen-glucose deprivation (OGD) damage in organotypic hippocampal slices is also made worse by UPF 1069 (10 µM). On the other hand, UPF 1069 reduces the harm caused by OGD in cultures of mixed cortical cells and exhibits strong neuroprotective activity at concentrations of 1 µM, which acts specifically on PARP-2, and 10 µM, which inhibits the activities of both PARP-1 and PARP-2[2].
ln Vivo	Oxygen-glucose deprivation (OGD)-induced CA1 pyramidal cell death in organotypic hippocampal slices is concentration-dependently exacerbated (up to 155%) when PARP-2 inhibition with UPF-1069 (0.01-1 mM) is applied. Elevated concentrations do not affect OGD injury, since they only affect PARP-1 and PARP-2. The administration of UPF-1069 (1-10 mM) to mouse mixed cortical cells exposed to OGD significantly mitigates post-ischaemic damage.
Enzyme Assay	Recombinant mouse PARP-2 and bovine PARP-1, which are readily available for purchase, are used to measure PARP activity. The enzymatic reaction is conducted in 100 µL of 50 mM Tris-HCl (pH 8.0) supplemented with 5 mM MgCl22, 2 mM dithiothreitol, 10 µg of sonicated calf thymus DNA, 0.2 µCi [adenine-2,8-3H]NAD, and recombinant enzyme PARP-1 or PARP-2 (0.03 U per sample). That is, in brief. The mixture is mixed and incubated at 37°C for one hour at varying concentrations of the potential inhibitors. After adding 1 mL of 10% trichloroacetic acid (w/v) to stop the reaction, the mixture is centrifuged. Following two washings in 1 mL of H2O, the pellets are resuspended in 1 mL of 0.1 M NaOH. Liquid scintillation spectrometry is used to measure the radioactivity incorporated from [adenine-2,8-3H]NAD into proteins[2].
Animal Protocol	0.01-1 mM Mouse
References	[1]. On the way to selective PARP-2 inhibitors. Design, synthesis, and preliminary evaluation of a series of isoquinolinone derivatives. ChemMedChem. 2008 Jun;3(6):914-23. [2]. Selective PARP-2 inhibitors increase apoptosis in hippocampal slices but protect cortical cells in models of post-ischaemic brain damage.

Solubility Data

Solubility (In Vitro)

DMSO: ~56 mg/mL (~200.5 mM) Water: <1 mg/mL Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (8.95 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 (8.95 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. Solubility in Formulation 3: 2% DMSO+30% PEG 300+5% Tween 80+ddH2O: 5mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.5805 mL	17.9025 mL	35.8051 mL
	5 mM	0.7161 mL	3.5805 mL	7.1610 mL
	10 mM	0.3581 mL	1.7903 mL	3.5805 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.