Rucaparib (formerly AG-014699 or PF-01367338; trade name: Rubraca) is a potent, tricyclic indole based, and orally bioavailable inhibitor of PARP (poly(ADP-Ribose) polymerase) with potential anticancer activity. In cell-free experiments, it inhibits PARP1 with a Ki of 1.4 nM. The US FDA approved rucaparib in December 2016 for the treatment of ovarian cancer in women. Rucaparib binds specifically to PARP1 and prevents PARP1 from repairing damaged DNA, which increases the number of breaks in DNA strands and encourages apoptosis and genomic instability. This could reverse tumor cell resistance to chemotherapy and radiation therapy and increase the cytotoxicity of agents that damage DNA.

Physicochemical Properties

Molecular Formula	C19H18FN3O.H3PO4
Molecular Weight	421.36
Exact Mass	421.12
CAS #	459868-92-9
Related CAS #	1859053-21-6 (camsylate); 283173-50-2; 459868-92-9(phosphate)
PubChem CID	9931953
Appearance	Light yellow solid powder
Boiling Point	625.2ºC at 760 mmHg
Flash Point	331.9ºC
LogP	2.77
Hydrogen Bond Donor Count	6
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	3
Heavy Atom Count	29
Complexity	515
Defined Atom Stereocenter Count	0
SMILES	O=P(O)(O)O.FC1=CC2=C3C(CCNC2=O)=C(C4=CC=C(CNC)C=C4)NC3=C1
InChi Key	FCCGJTKEKXUBFZ-UHFFFAOYSA-N
InChi Code	InChI=1S/C19H18FN3O.H3O4P/c1-21-10-11-2-4-12(5-3-11)18-14-6-7-22-19(24)15-8-13(20)9-16(23-18)17(14)15;1-5(2,3)4/h2-5,8-9,21,23H,6-7,10H2,1H3,(H,22,24);(H3,1,2,3,4)
Chemical Name	6-fluoro-2-[4-(methylaminomethyl)phenyl]-3,10-diazatricyclo[6.4.1.04,13]trideca-1,4,6,8(13)-tetraen-9-one;phosphoric acid
Synonyms	AG014699; PF-01367338; AG 014699; PF 01367338; AG-014699; PF01367338; AG-14447; AG 14447; AG14447; Trade name: Rubraca; Rucaparib
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	PARP-1 ( Ki = 1.4 nM ); PARP-2/3
ln Vitro	In vitro activity: Rucaparib exhibits a greater inhibition of cellular PARP in LoVo and SW620 cells and is a potent inhibitor of purified full-length human PARP-1. Additionally, eight additional PARP domains, namely PARP2, 3, 4, 10, 15, 16, TNKS1 and TNKS2, are detectably bound by rucaparib. [1] [2] Rucaparib causes radiosensitization independent of SSB repair inhibition, as it inhibits NF-κB activation downstream. Rucaparib has the potential to bypass the toxicity associated with traditional NF-κB inhibitors by targeting NF-κB that is activated by DNA damage, all while maintaining other essential inflammatory functions. [3] Rucaparib, at a concentration of 1 μM, inhibits PARP-1 activity in permeabilized D283Med cells by 97.1%. [4]
ln Vivo	Rucaparib is not toxic, but in DNA repair protein-competent D384Med xenografts, it dramatically increases temozolomide-induced TGD. Additionally, pharmacokinetic studies demonstrate that rucaparib is found in brain tissue, suggesting rucaparib's potential for intracranial cancer treatment. [4] The cytotoxicity of temozolomide and topotecan in NB-1691, SH-SY-5Y, and SKNBE (2c) cells is markedly enhanced by rucaparib. In NB1691 and SHSY5Y xenografts, rucaparib improves the antitumor activity of temozolomide and shows total and prolonged tumor regression. [5]
Enzyme Assay	The amount of [32P]NAD+ incorporation-induced inhibition of human full-length recombinant PARP-1 is measured. With a PhosphorImager, the amount of [32P]ADP-ribose added to acid-insoluble material is measured. The nonlinear regression analysis is used to calculate Ki.
Cell Assay	The density of medulloblastoma cell lines is plated at 1 × 103, 3 × 103, and 3 × 103 in 96-well plates, in that order. Cells are seeded and exposed to different concentrations of temozolomide with or without 0.4 μM Rucaparib at 24, 48, or 72 hours later (D283Med and D425Med). Utilizing an XTT cell proliferation kit assay, cell viability is assessed following 3 days (D425Med and D384Med) or 5 days (D283Med) of culture. In comparison to DMSO or 0.4 μM Rucaparib-alone controls, the percentage of cell growth is given. One can compute the growth inhibition by 50% (GI50) of temozolomide, either in isolation or in conjunction with Rucaparib. The ratio of the GI50 of temozolomide in the presence of Rucaparib to the GI50 of temozolomide alone is known as the potentiation factor 50 (PF50).
Animal Protocol	Female athymic nude mice, implanted SW620 colorectal tumor cells (1 × 107 cells per animal) s.c. 0.1 mg/kg in combination with Temozolomide (p.o., 200 mg/kg), 0.05, 0.15, and 0.5 mg/kg in combination with Temozolomide (p.o., 68 mg/kg) or 10 mg/kg IP, single dose for 0.1 mg/kg and 10 mg/kg, five daily doses for 0-0.5 mg/kg
ADME/Pharmacokinetics	Absorption Rucaparib exhibits a linear pharmacokinetic profile over the dose range from 240 mg to 840 mg twice daily. The mean (coefficient of variation [CV]) steady-state rucaparib Cmax is 1940 ng/mL (54%) and AUC0-12h is 16900 h x ng/mL (54%) at the approved recommended dosage. The mean AUC accumulation ratio is 3.5 to 6.2 fold. The median Tmax at the steady state is 1.9 hours, with a range of 0 to 5.98 hours at the approved recommended dosage. The mean absolute bioavailability is 36%, with a range of 30 to 45%. A high-fat meal increased Cmax and AUC0-24h by 20% and 38%, respectively. The Tmax was delayed by 2.5 hours. Route of Elimination Following a single oral dose of radiolabeled rucaparib, unchanged rucaparib accounted for 64% of the radioactivity. Rucaparib accounted for 45% and 95% of radioactivity in urine and feces, respectively. Volume of Distribution The mean (coefficient of variation) apparent volume of distribution is 2300 L (21%). Clearance The mean (coefficient of variation) apparent total clearance at steady state is 44.2 L/h (45%). Metabolism / Metabolites In vitro, rucaparib is primarily metabolized by CYP2D6 and, to a lesser extent, by CYP1A2 and CYP3A4. In addition to CYP-based oxidation, rucaparib also undergoes N-demethylation, N-methylation, and glucuronidation. In one study, seven metabolites of rucaparib were identified in plasma, urine, and feces. Biological Half-Life The mean (coefficient of variation) terminal elimination half-life is 26 (39%) hours.
Toxicity/Toxicokinetics	Hepatotoxicity In large clinical trials of rucaparib, abnormalities in routine liver tests were common; serum ALT elevations arising in 74% with values above 5 times the upper limit of normal (ULN) in 13%. Despite the frequency of serum enzyme elevations during therapy in clinical trials, there were no reports of hepatitis with jaundice or liver failure. Subsequent to its approval and more wide scale use, there have been no published reports of clinically apparent liver injury attributed to rucaparib. Thus, rucaparib is a frequent cause of serum enzyme elevations, but has not been linked to significant hepatotoxicity. Likelihood score: E* (unproven but suspected cause of clinically apparent liver injury). Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation No information is available on the clinical use of rucaparib during breastfeeding. The manufacturer recommends that breastfeeding be discontinued during rucaparib therapy and for 2 weeks after the last dose. ◉ Effects in Breastfed Infants Relevant published information was not found as of the revision date. ◉ Effects on Lactation and Breastmilk Relevant published information was not found as of the revision date. Protein Binding Rucaparib is 70% bound to human plasma proteins _in vitro_. Rucaparib preferentially distributed to red blood cells with a blood-to-plasma concentration ratio of 1.8.
References	[1]. Mol Cancer Ther . 2007 Mar;6(3):945-56. [2]. SciBX 5(13). [3]. Oncogene . 2012 Jan 12;31(2):251-64. [4]. Br J Cancer . 2010 Nov 9;103(10):1588-96. [5]. Clin Cancer Res . 2009 Feb 15;15(4):1241-9.
Additional Infomation	Rucaparib Phosphate is the phosphate salt form of rucaparib, an orally bioavailable tricyclic indole and inhibitor of poly(ADP-ribose) polymerases (PARPs) 1 (PARP1), 2 (PARP2) and 3 (PARP3), with potential chemo/radiosensitizing and antineoplastic activities. Upon administration, rucaparib selectively binds to PARP1, 2 and 3 and inhibits PARP-mediated DNA repair. This enhances the accumulation of DNA strand breaks, promotes genomic instability and induces cell cycle arrest and apoptosis. This may enhance the cytotoxicity of DNA-damaging agents and reverse tumor cell resistance to chemotherapy and radiation therapy. PARPs are enzymes activated by single-strand DNA breaks that catalyze the post-translational ADP-ribosylation of nuclear proteins, which induces signaling and the recruitment of other proteins to repair damaged DNA. The PARP-mediated repair pathway plays a key role in DNA repair and is dysregulated in a variety of cancer cell types.

Solubility Data

Solubility (In Vitro)

DMSO:~84 mg/mL (~199.4 mM) Water: <1 mg/mL Ethanol: <1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.93 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. 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 (5.93 mM) (saturation unknown) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. 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: ≥ 2.17 mg/mL (5.15 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 21.7 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 4: ≥ 2.17 mg/mL (5.15 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 21.7 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 5: ≥ 2.17 mg/mL (5.15 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 21.7 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. Solubility in Formulation 6: ≥ 0.5 mg/mL (1.19 mM) (saturation unknown) in 1% DMSO 99% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 7: 30% propylene glycol, 5% Tween 80, 65% D5W: 30mg/mL  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.3733 mL	11.8663 mL	23.7327 mL
	5 mM	0.4747 mL	2.3733 mL	4.7465 mL
	10 mM	0.2373 mL	1.1866 mL	2.3733 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.