Rucaparib CAMSYLATE (AG014699;AG14447; PF01367338; trade name:Rubraca), theCAMSYLATE salt of rucaparib, is a potent PARP inhibitor approved by FDA for the treatment of ovarian cancer. In a test without cells, it inhibits PARP1 with a Ki of 1.4 nM.
Physicochemical Properties
| Molecular Formula | C29H34FN3O5S |
| Molecular Weight | 555.67 |
| Exact Mass | 555.22 |
| Elemental Analysis | 555.670 Elemental Analysis: |
| CAS # | 1859053-21-6 |
| Related CAS # | 459868-92-9 (phosphate); 1859053-21-6 (Rucaparib camsylate); 283173-50-2 |
| PubChem CID | 121490161 |
| Appearance | Light yellow to gray solid powder |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 5 |
| Heavy Atom Count | 39 |
| Complexity | 869 |
| Defined Atom Stereocenter Count | 2 |
| SMILES | S(C([H])([H])[C@@]12C(C([H])([H])[C@@]([H])(C([H])([H])C1([H])[H])C2(C([H])([H])[H])C([H])([H])[H])=O)(=O)(=O)O[H].FC1=C([H])C2C(N([H])C([H])([H])C([H])([H])C3=C(C4C([H])=C([H])C(C([H])([H])N([H])C([H])([H])[H])=C([H])C=4[H])N([H])C(=C1[H])C3=2)=O |
| InChi Key | INBJJAFXHQQSRW-STOWLHSFSA-N |
| InChi Code | InChI=1S/C19H18FN3O.C10H16O4S/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;1-9(2)7-3-4-10(9,8(11)5-7)6-15(12,13)14/h2-5,8-9,21,23H,6-7,10H2,1H3,(H,22,24);7H,3-6H2,1-2H3,(H,12,13,14)/t;7-,10-/m.1/s1 |
| Chemical Name | [(1S,4R)-7,7-dimethyl-2-oxo-1-bicyclo[2.2.1]heptanyl]methanesulfonic acid;6-fluoro-2-[4-(methylaminomethyl)phenyl]-3,10-diazatricyclo[6.4.1.04,13]trideca-1,4,6,8(13)-tetraen-9-one |
| Synonyms | AG014699 camsylate; PF-01367338; AG 014699; PF 01367338 camsylate; AG-014699; PF01367338; AG-14447 camsylate; AG 14447; Rucaparib monocamsylate; Rucaparib (monocamsylate); Rucaparib (Camsylate); rucaparib camphorsulfonate; CO-338; PF-1367338-BW; AG14447 camsylate; Trade name: Rubraca |
| 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; PARP-3 |
| ln Vitro |
In vitro activity: Rucaparib (AG014699) camsylate is a possible N-demethylation metabolite of AG14644[1]. Rucaparib (0.1, 1, 10, 100 μM; 24 hours) camsylate is cytotoxic; in Capan-1 (BRCA2 mutant) cells, its LC50 is 5 μM, while in MX-1 (BRCA1 mutant) cells, it is only 100 nM[2]. Rucaparib camsylate induces radiosensitization independent of SSB repair inhibition, as it results from downstream inhibition of NF-κB activation. Without impairing other essential inflammatory functions, rucaparib camsylate can target NF-κB that is activated by DNA damage and overcome the toxicity seen with classical NF-κB inhibitors[5]. Rucaparib camsylate inhibits PARP-1 activity in permeabilized D283Med cells by 97.1% at a concentration of 1 μM[6]. |
| ln Vivo | Rucaparib camsylate and AG14584 significantly increase Temozolomide toxicity. Temozolomide-induced body weight loss is markedly enhanced by rucaparib (1 mg/kg) camsylate. The temozolomide-induced tumor growth delay is increased by 50% when rucaparib (0.1 mg/kg) camsylate is used[1]. Rucaparib (10 mg/kg intraperitoneally or 50–150 mg/kg po; daily, five days a week, for six weeks) There is one complete tumor regression and two persistent partial tumor regressions as a result of camsylate's strong tumor growth inhibition[2]. The most potent antitumor effect is achieved with three complete regressions when rutaparib (150 mg/kg; p.o. ; once weekly for 6 weeks or three times weekly for 6 weeks) is used for camsylate[2]. Rucaparib camsylate in NB1691 and SHSY5Y xenografts shows complete and sustained tumor regression and improves the antitumor activity of temozolomide[6]. |
| Enzyme Assay | It is measured how much [32P]NAD+incorporation inhibits human full-length recombinant PARP-1. A PhosphorImager is used to quantify the [32P]ADP-ribose that is integrated into acid-insoluble material. Nonlinear regression analysis is used to calculate Kiis. |
| Cell Assay | The following day, the cells were treated with escalating drug concentrations after being seeded into 24-well plates (2,500–4000 cells/well). After 72–96 hours, cell viability was evaluated by adding CellTiter-Glo reagent and utilizing a plate reader to measure luminescence. Cells were seeded in 6-well plates in triplicate (500–4000 cells/well) for clonogenic survival assays. Following plating, cells were given a drug treatment 16–18 hours later and were left to grow for 14 days. |
| Animal Protocol |
CD-1 nude mice bearing established D283Med xenografts; Dissolved in saline;1 mg/kg;One or four daily by i.p. Determination of Antitumor Activity In vivo[1] Female athymic nude mice (CD1 nu/nu) used for antitumor studies were maintained and handled in isolators under specific pathogen-free conditions. We implanted SW620 colorectal tumor cells (1 × 107 cells per animal) s.c. into one flank of each mouse, treated the mice (five animals per group) when tumors were palpable (10–12 days after implantation), and monitored tumor growth using two-dimensional caliper measurements. Tumor volume was calculated using the equation a2 × b / 2, where a is the smallest measurement and b is the largest. Data are presented as median relative tumor volumes (RTV), defined as the calculated tumor volume divided by the calculated tumor volume on the initial day of treatment (day 0). Thus, on day 0, the RTV value is 1 and RTV4 is when the tumor is four times as large as its initial value. Single-Dose Studies. [1] We administered a single dose of temozolomide p.o. as a suspension in saline at 200 mg/kg either alone or in combination with a single i.p. administration of PARP inhibitor administered at 0.1 [AG14447 and MS-AG14644 (equivalent to 0.078 mg/kg free AG14644 only)], 1.0, and 10 mg/kg (for the mesylate salts equivalent to 0.79 and 7.9 mg/kg free AG14451 and AG14452 and 0.78 and 7.8 free AG14531 and AG14644). Control animals were treated with either normal saline p.o. and i.p or normal saline p.o and PARP inhibitor 10 mg/kg i.p. Five Daily Dosing Studies. [1] We treated animals with five daily doses of temozolomide administered p.o. as a suspension in saline at 68 mg/kg either alone or in combination with a five daily i.p. administrations of PARP inhibitor at 0.05, 0.15, and 0.5 mg/kg AG14447; 0.15 and 0.5 mg/kg MS-AG14644 (equivalent to 0.12 and 0.39 mg/kg free AG14644); 1.5, 5, and 15 mg/kg AG14361; and 5 mg/kg AG14452. Control animals were treated with either normal saline p.o. and i.p. or normal saline p.o and PARP inhibitor at the higher dose (0.5, 5, or 15 mg/kg, depending on the compound studied) i.p. Tissue Distribution[1] We administered AG14361, AG14452, or AG14447 (10 mg/kg i.p.) to mice (three animals per group) bearing SW620 xenografts (∼10 × 10 mm). After 120 min, the animals were bled by cardiac puncture, under general anesthesia, the tumor was removed and snap frozen on liquid nitrogen. Plasma was removed and stored at −20°C. The concentrations of PARP inhibitor in acetonitrile-treated plasma and homogenized tumor were measured using reverse-phase high-pressure liquid chromatography (isocratic mobile phase: 40% acetonitrile in 0.1% ammonium formate, Hypersil BDS 3 μm 4.6 × 250 mm column, Waters Alliance 2690 high-pressure liquid chromatography; Waters, Elstree, Herts, United Kingdom) by the method of addition. |
| 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]. Preclinical selection of a novel poly(ADP-ribose) polymerase inhibitor for clinical trial. Mol Cancer Ther, 2007, 6(3), 945-956. [2]. Tumour cell retention of rucaparib, sustained PARP inhibition and efficacy of weekly as well as daily schedules. Br J Cancer. 2014 Apr 15;110(8):1977-84. [3]. Rucaparib: A Review in Ovarian Cancer. Target Oncol. 2019 Apr;14(2):237-246. [4]. Hexose-6-phosphate dehydrogenase blockade reverses prostate cancer drug resistance in xenograft models by glucocorticoid inactivation. Sci Transl Med. 2021 May 26;13(595):eabe8226. [5]. NF-κB mediates radio-sensitization by the PARP-1 inhibitor, AG-014699. Oncogene, 2012, 31(2), 251-264. [6]. Inhibition of poly(ADP-ribose) polymerase-1 enhances temozolomide and topotecan activity against childhood neuroblastoma. Clin Cancer Res, 2009, 15(4), 1241-1249. |
| Additional Infomation |
Rucaparib camsylate is a camphorsulfonate salt obtained by reaction of rucaparib with one molar equivalent of (1S,4R)-camphorsulfonic acid. It is an inhibitor of poly (ADP-ribose) polymerase and is used as monotherapy for advanced ovarian cancer and deleterious germline or somatic BRCA mutation. It has a role as an EC 2.4.2.30 (NAD(+) ADP-ribosyltransferase) inhibitor. It is a camphorsulfonate salt and an azepinoindole. It contains a (S)-camphorsulfonate and a rucaparib(1+). Rucaparib Camsylate is the camsylate 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. See also: Rucaparib (has active moiety). Drug Indication Treatment of fallopian tube cancer , Treatment of ovarian cancer , Treatment of primary peritoneal cancer , Treatment of prostate malignant neoplasms |
Solubility Data
| Solubility (In Vitro) |
DMSO: ≥ 100 mg/mL Water: N/A Ethanol: N/A |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.74 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 20.8 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: ≥ 2.08 mg/mL (3.74 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 20.8 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 3: ≥ 2.08 mg/mL (3.74 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 20.8 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 | 1.7996 mL | 8.9981 mL | 17.9963 mL | |
| 5 mM | 0.3599 mL | 1.7996 mL | 3.5993 mL | |
| 10 mM | 0.1800 mL | 0.8998 mL | 1.7996 mL |