PARP-1 ( Ki = 1.4 nM ); PARP-2; PARP-3
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].

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].

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Uptake of rucaparib into SW620 human colorectal cancer cells is carrier-mediated and follows Michaelis-Menten kinetics (Km = 8.4 ± 1.2 µM, Vmax = 469 ± 22 pmol per 10^6 cells per 10 min). [2]
Rucaparib accumulates within cells against a concentration gradient, reaching intracellular steady-state levels >10 times higher than the extracellular concentration within 30 minutes of exposure to 400 nM drug. Following a 30-minute pulse, efflux is biphasic, with a rapid initial loss followed by very slow efflux; intracellular concentrations remain >10-fold above the original extracellular level 2 hours after drug removal. The half-life for exponential decline in intracellular concentration over 24 hours is approximately 20 minutes. [2]
A 30-minute pulse of 400 nM rucaparib results in sustained PARP inhibition ≥50% for at least 72 hours in SW620, Capan-1 (BRCA2 mutant), and MX-1 (BRCA1 mutant) cells. In Capan-1 and MX-1 cells, PARP activity was still below baseline levels 1 week after drug removal. [2]
Rucaparib is cytotoxic to homologous recombination-deficient cells. The LC50 (concentration reducing colony formation by 50%) after a 24-hour exposure was 5 µM in Capan-1 cells and 100 nM in MX-1 cells. [2]
The carboxylic acid metabolite of rucaparib inhibits PARP activity in permeabilized cells with an IC50 of ~550 nM but does not inhibit PARP in intact cells, suggesting poor membrane permeability. [2]

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].

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In mice bearing Capan-1 (BRCA2 mutant pancreatic cancer) xenografts, a single oral dose of rucaparib (150 mg/kg) inhibited tumour PARP activity by >90% for up to 7 days. A single intraperitoneal (i.p.) dose (10 mg/kg) also caused prolonged inhibition (>70% at 24h, with only modest recovery to ~30% of baseline by 48h and no further recovery over 4 days). [2]
Weekly oral administration of rucaparib (150 mg/kg once per week for 6 weeks) was as effective as, or more effective than, daily i.p. administration (10 mg/kg, 5 days per week for 6 weeks) in delaying the growth of Capan-1 xenografts. The weekly schedule resulted in three complete tumour regressions out of ten mice. Daily oral administration (150 mg/kg, 5 days per week) had equivalent efficacy to the i.p. schedule. [2]
Rucaparib showed no significant antitumour activity against slowly growing MX-1 (BRCA1 mutant breast cancer) xenografts, despite achieving >80% PARP inhibition in the tumours 24 hours after a single 150 mg/kg oral dose. [2]
PARP inhibition appears more sustained in tumour tissue compared to normal tissues. While tumour PARP activity was suppressed for a week, activity in the liver recovered within 24 hours, and activity in peripheral blood mononuclear cells (PBMCs) recovered to 35-40% of control by 24 hours post-dose. [2]

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.

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PARP activity was measured using a validated immunoblot assay. Briefly, tissues (tumour, brain, liver) were homogenized in isotonic buffer. For the assay, PARP activity in permeabilized cells (e.g., PBMCs) or tissue homogenates was maximally stimulated by incubation with a double-stranded oligonucleotide (mimicking DNA breaks) in the presence of excess NAD+ (350 µM). The amount of poly(ADP-ribose) (PAR) polymer formed was quantified by immunoblotting using an anti-PAR antibody, with reference to a standard curve of purified PAR. Results were expressed as pmol PAR per 10^6 cells or per mg protein. [2]

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.

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Drug Uptake and Retention (Radiochemical Assay): Exponentially growing cells were incubated with [¹⁴C]rucaparib and a non-cell-permeant tracer (e.g., [³H]sucrose/inulin) to correct for extracellular fluid. At specified times, cells were rapidly separated from medium by centrifugation through a silicone oil layer into a KOH solution. Radioactivity in the cell pellet was determined by dual-label scintillation counting, and intracellular drug concentration was calculated after correcting for trapped extracellular drug. For retention studies, cells were pulsed with the drug, washed, and incubated in fresh medium for varying periods before processing. [2]
Clonogenic Survival Assay: Exponentially growing Capan-1 or MX-1 cells were seeded at low density in multi-well dishes. After attachment, cells were exposed to increasing concentrations of rucaparib for 24 hours, then the medium was replaced with drug-free medium. Colonies were allowed to form for 10-14 days, stained, counted, and survival relative to untreated controls was calculated. [2]
Cellular PARP Activity Assay: Following drug treatments (e.g., a 30-minute pulse), cells were washed, harvested, and cryopreserved. PARP activity was later measured in permeabilized cells using the immunoblot-based assay described in the "Enzyme Assay" section. [2]

CD-1 nude mice bearing established D283Med xenografts; Dissolved in saline;1 mg/kg;One or four daily by i.p.

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\n \nDetermination of Antitumor Activity In vivo[1]
\nFemale 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. \n

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\nSingle-Dose Studies. [1]
\nWe 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. \n

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\nFive Daily Dosing Studies. [1]
\nWe 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.\n

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\nTissue Distribution[1]
\nWe 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.\n

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\nXenograft Establishment: Female CD-1 nude mice (10-12 weeks old) were used. Capan-1 cells were implanted subcutaneously in the flank. MX-1 cells were implanted in a 1:1 mixture of growth factor-reduced basement membrane matrix and medium. [2]
\nPharmacokinetics and Tissue Distribution Study: Mice bearing established Capan-1 tumours (approx. 25-50 mm³) received a single dose of rucaparib intraperitoneally (10 mg/kg) or orally (50, 100, or 150 mg/kg). The drug was dissolved in sterile deionized water. Groups of mice (n=3 per time point) were euthanized at 0.5, 4, 24, 48, 72, and 168 hours post-dose. Blood (for plasma), tumours, and brains were collected, snap-frozen, and stored for analysis of drug concentration (by HPLC) and PARP activity. [2]
\nEfficacy Study: Mice bearing palpable Capan-1 tumours (≥5x5 mm) were randomized into treatment groups (n=8-10). Rucaparib was administered according to various schedules for 6 weeks: vehicle control; 10 mg/kg i.p. daily for 5 days per week; 50 or 150 mg/kg p.o. daily for 5 days per week; 150 mg/kg p.o. once per week; 150 mg/kg p.o. three times per week; or 150 mg/kg p.o. daily for 5 days every 3 weeks. Tumour dimensions were measured regularly, and volume was calculated. Mice were monitored for weight loss and humanely killed if tumours reached a predefined size limit or if signs of stress occurred. [2]

Absorption
Rucapparib exhibits linear pharmacokinetic characteristics at twice-daily doses ranging from 240 mg to 840 mg. At the approved recommended dose, the mean steady-state Cmax (coefficient of variation [CV]) of rucapaparib is 1940 ng/mL (54%), and the AUC0–12h is 16900 h × ng/mL (54%). The mean AUC cumulative ratio is 3.5 to 6.2 times. At the approved recommended dose, the median steady-state Tmax is 1.9 hours, ranging from 0 to 5.98 hours. The mean absolute bioavailability is 36%, ranging from 30% to 45%. High-fat meals increased Cmax and AUC0–24h by 20% and 38%, respectively, and delayed Tmax by 2.5 hours.
Elimination Pathway
After a single oral administration of radiolabeled rucapanib, 64% of the total radioactivity is unmetabolized. 45% and 95% of the total radioactivity are found in urine and feces, respectively.
Volume of Distribution
The mean apparent volume of distribution (coefficient of variation) is 2300 L (21%).
Clearance
The mean apparent total clearance at steady state (coefficient of variation) is 44.2 L/h (45%).
Metabolites/Metabolites
In vitro studies have shown that rucapanib is primarily metabolized by CYP2D6, followed by CYP1A2 and CYP3A4. In addition to CYP-mediated oxidation, rucapanib also undergoes N-demethylation, N-methylation, and glucuronidation. In one study, researchers identified seven rucapanib metabolites in plasma, urine, and feces.

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Biological half-life
The mean (coefficient of variation) terminal elimination half-life is 26 (39%) hours.

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Rucapanib has oral bioavailability in mice. Following a single oral administration (50-150 mg/kg), the parent drug can be detected in plasma for up to 4-48 hours, depending on the dose. [2]
Rucapanib has significantly higher concentrations and longer retention times in tumor tissues compared to plasma. Four hours after administration, the drug concentration in tumors is 10 times higher than the plasma concentration at the same time, and the drug can be detected in tumors for up to 3 days after oral administration. [2]
The drug has limited uptake into the brain, with drug concentrations in brain tissue ≤ 10% of plasma concentrations. [2] A carboxylic acid metabolite was detected in both plasma and tumors, with plasma concentrations comparable to or higher than the parent drug, but lower concentrations in tumors. The metabolite exhibits weak PARP inhibitory activity (IC50 of approximately 550 nM in cell-free assays) but does not inhibit PARP in intact cells. [2] Oral bioavailability in mice is similar to that in humans, and it is rapidly eliminated from plasma. A linear relationship between dose and drug concentration was observed. [2]

Hepatotoxicity
In large clinical trials of rucaparib, abnormalities in routine liver function tests were common; 74% of patients experienced elevated serum ALT, with 13% of these patients having ALT values exceeding 5 times the upper limit of normal (ULN). Although elevated serum enzymes were common during treatment in clinical trials, no reports of hepatitis with jaundice or liver failure were observed. Since the approval and wider use of rucaparib, no clinically significant cases of liver injury have been published. Therefore, rucaparib is a common cause of elevated serum enzymes, but it has not been found to be associated with significant hepatotoxicity.
Probability score: E (Unproven but suspected cause of clinically significant liver injury).
Pregnancy and Lactation Effects
◉Overview of Use During Lactation
There is currently no information regarding the clinical use of rucaparib during lactation. The manufacturer recommends discontinuing breastfeeding during treatment with rucaparib and for 2 weeks after the last dose.
◉ Effects on breastfed infants
No relevant published information was found as of the revision date.
◉ Effects on lactation and breast milk
No relevant published information was found as of the revision date.

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Protein binding
In vitro studies showed that rucaparib binds to human plasma proteins in 70% of cases. Rucaparib preferentially distributes to erythrocytes, with a plasma concentration ratio of 1.8.

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In efficacy studies in mice carrying Capan-1 or MX-1 xenografts, treatment with rucaparib at different dosing regimens (daily or weekly oral doses up to 150 mg/kg) did not cause any significant weight loss (minimum weight loss ≤4%). [2]
The manuscript mentions that rucaparib monotherapy at a dose of 50 mg/kg repeated every 5 days for 6 months was non-toxic in mice. The maximum tolerated dose was lower when used in combination with temozolomide. [2] Clinical data (cited but not from this study) indicate that tolerated doses of PARP inhibitor monotherapy can cause grade 3/4 myelosuppression in combination chemotherapy studies. Neurological side effects (e.g., drowsiness) have been reported with PARP inhibitor monotherapy. The low brain penetration of rucapranib observed in mice may reduce the incidence of such cognitive/neurological side effects. [2]

[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.

Rucaparib camphor sulfonate is a camphor sulfonate produced by reacting rucaparib with one molar equivalent of (1S,4R)-camphor sulfonic acid. It is an inhibitor of poly(ADP-ribose) polymerase and is used to treat advanced ovarian cancer and ovarian cancer carrying pathogenic germline or somatic BRCA mutations. It is an EC 2.4.2.30 (NAD(+) ADP-ribosyltransferase) inhibitor. It is a camphor sulfonate and azaheptanoid indole compound. It contains one (S)-camphor sulfonate and one rucaparib (1+). Rucaparib camphor sulfonate is the camphor sulfonate form of rucaparib, a highly bioavailable tricyclic indole compound that is also an inhibitor of poly(ADP-ribose) polymerases (PARP) 1 (PARP1), 2 (PARP2), and 3 (PARP3), with potential chemosensitizing/radiosensitizing and antitumor activity. After administration, rucaparib selectively binds to PARP1, 2, and 3, inhibiting 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 radiotherapy. PARP is an enzyme activated by single-strand DNA breaks, catalyzing post-translational ADP ribosylation of nucleoproteins, thereby inducing signal transduction and recruiting other proteins to repair damaged DNA. PARP-mediated repair pathways play a crucial role in DNA repair and are dysregulated in various cancer cell types.
See also: Rucaparib (with active moiety).
Drug Indications

Treatment of fallopian tube cancer, treatment of ovarian cancer, treatment of primary peritoneal cancer, treatment of prostate cancer
Rucaparib
(AG-014699) is the first PARP inhibitor to enter clinical trials for cancer treatment. [2]
The sustained PARP inhibition observed after short-term drug exposure was attributed to carrier-mediated cellular uptake, accumulation against the concentration gradient, and affinity binding to the enzyme (Ki ~1.4 nM), rather than PARP "capture" on DNA. [2]
Long-term inhibition and retention of tumor PARP supports the clinical exploration of intermittent dosing regimens (e.g., once weekly), which may potentially reduce toxicity or improve ease of use while maintaining efficacy. [2]
In this study, MX-1 xenografts lacked efficacy, although in vitro studies showed sensitivity to the drug, which may be related to slow tumor growth or higher baseline PARP activity in the tumor in this model. [2]

C29H34FN3O5S

555.67

555.22

555.670 Elemental Analysis:

1859053-21-6

459868-92-9 (phosphate); 1859053-21-6 (Rucaparib camsylate); 283173-50-2

121490161

Light yellow to gray solid powder

4

7

5

39

869

2

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

INBJJAFXHQQSRW-STOWLHSFSA-N

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

[(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

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

2934.99.9001

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.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ≥ 100 mg/mL
Water: N/A
Ethanol: N/A

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.

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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.

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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.

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 (Please use freshly prepared in vivo formulations for optimal results.)