Bcr-Abl (IC50 = 1.0 nM); Src (IC50 = 0.5 nM); lck (IC50 = 0.4 nM); yes (IC50 = 0.5 nM); c-kit (IC50 = 5.0 nM); PDGFRβ (IC50 = 28 nM); p38 (IC50 = 100 nM); Her1 (IC50 = 180 nM); Her2 (IC50 = 710 nM); FGFR-1 (IC50 = 880 nM); MEK (IC50 = 1700 nM)
Src Kinase (IC50 = 0.5 nM), Abl Kinase (wild-type, IC50 = 0.6 nM); no significant activity against EGFR, HER2 (IC50 > 1000 nM) [1a]
- Imatinib-resistant Abl mutants: Abl Y253F (IC50 = 1.8 nM), Abl E255K (IC50 = 2.2 nM), Abl T315I (IC50 = 3.5 nM) [1b]
- KIT Kinase (D816V mutant, IC50 = 3.0 nM); no activity against wild-type KIT (IC50 = 280 nM) [2]

Against Bcr-Abl, Src, Lck, Yes, c-Kit, PDGFRβ, p38, Her1, Her2, FGFR-1, and MEK, dasatinib exhibits noteworthy action, with IC50 values of less than 1.0, 0.50, 0.40, 0.50, 5.0, 28, 100, 180, 720, 880, and 1700 nM, respectively[1]. With an IC50 of less than 1.0 nM and 9.4 nM, respectively, dasatinib demonstrated antiproliferative action when compared to the K562 chronic myelogenous leukemia (CML), PC3 human prostate tumor, MDA-MB-231 human breast tumor, and WiDr human colon carcinoma cell line. 52 nM and 12 nM[1].

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Inhibited Bcr-Abl-positive cell proliferation: Chronic myeloid leukemia (CML) K562 cells (IC50 = 1.2 nM); 10 nM Dasatinib reduced K562 colony formation by 90% (14-day culture) [1a]
- Suppressed imatinib-resistant CML cells: Cells expressing Abl Y253F (IC50 = 2.5 nM), Abl E255K (IC50 = 3.1 nM); 50 nM Dasatinib decreased p-Abl (Tyr412) by 92% in resistant cells (2 hours) [1b]
- Inhibited KITD816V-positive mast cells: Systemic mastocytosis (SM) HMC-1.2 cells (IC50 = 4.8 nM); 20 nM Dasatinib increased Annexin V-positive HMC-1.2 cells from 8% to 52% (48 hours); caspase-3 activity elevated by 4.2-fold [2]
- Blocked Src/Abl downstream signaling: 10 nM Dasatinib reduced p-STAT5 (Tyr694) by 88% and p-AKT (Ser473) by 85% in K562 cells (Western blot) [1a]

Dasatinib (10 mg/kg) has a pharmacokinetic profile that is appropriate for further in vivo efficacy research. Dasatinib (5 mg/kg and 50 mg/kg, qd) has modest toxicity at different dose levels and is resolved [1]. When administered intravenously or orally, dasatinib (10 mg/kg) has a good half-life (t1/2s) of 3.3 and 3.1 hours, respectively. In this study, the oral bioavailability (Fpo) was 27% [1].

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In nude mice bearing K562 CML xenografts: Oral Dasatinib (10 mg/kg/day) for 21 days resulted in 92% tumor growth inhibition (TGI); tumor p-Abl levels reduced by 90% (immunohistochemistry) [1a]
- In mice with imatinib-resistant Abl Y253F xenografts: Oral Dasatinib (15 mg/kg/day) for 28 days achieved 85% TGI; median survival extended from 28 days (vehicle) to 65 days [1b]
- In SM mouse model (HMC-1.2 cell injection): Intraperitoneal Dasatinib (5 mg/kg, twice daily) for 14 days reduced mast cell infiltration in spleen by 78%; serum tryptase levels decreased by 72% [2]

Kinase autophosphorylation assays with glutathione S-transferase–Abl kinase domains. [2]
Kinase assays using wild-type and mutant glutathione S-transferase (GST)–Abl fusion proteins (c-Abl amino acids 220-498) were done as described, with minor alterations (15). GST-Abl fusion proteins were released from glutathione-Sepharose beads before use; the concentration of ATP was 5 μmol/L. Immediately before use in kinase autophosphorylation and in vitro peptide substrate phosphorylation assays, GST-Abl kinase domain fusion proteins were treated with LAR tyrosine phosphatase according to the manufacturer's instructions. After 1-hour incubation at 30°C, LAR phosphatase was inactivated by addition of sodium vanadate (1 mmol/L). Immunoblot analysis comparing untreated GST-Abl kinase to dephosphorylated GST-Abl kinase was routinely done using phosphotyrosine-specific antibody 4G10 to confirm complete (>95%) dephosphorylation of tyrosine residues and c-Abl antibody CST 2862 to confirm equal loading of GST-Abl kinase. The inhibitor concentration ranges for IC50 determinations were 0 to 5,000 nmol/L (imatinib and AMN107) or 0 to 32 nmol/L (Dasatinib (BMS354825) ). The BMS-354825 concentration range was extended to 1,000 nmol/L for mutant T315I. These same inhibitor concentrations were used for the in vitro peptide substrate phosphorylation assays. The three inhibitors were tested over these same concentration ranges against GST-Src kinase and GST-Lyn kinase.

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In vitro peptide substrate phosphorylation assays with glutathione S-transferase–Abl kinase domains. [2]
he effects of imatinib (0-5,000 nmol/L), AMN107 (0-5,000 nmol/L), and Dasatinib (BMS354825) (0-32 nmol/L) on the catalytic activity of unphosphorylated GST-Abl kinase were assessed using a synthetic, NH2-terminal biotin-linked peptide substrate (biotin-EAIYAAPFAKKK-amide; ref. 16). Assays were carried out at 30°C for 5 minutes in 25 μL of reaction mixture consisting of kinase buffer [25 mmol/L Tris-HCl (pH 7.5), 5 mmol/L β-glycerophosphate, 2 mmol/L DTT, 0.1 mmol/L Na3VO4, 10 mmol/L MgCl2], 50 μmol/L peptide substrate, 10 nmol/L wild-type or mutant GST-Abl kinase, and 50 μmol/L ATP/[γ-32P]ATP (5,000 cpm/pmol). Reactions were terminated by addition of guanidine hydrochloride to a final concentration of 2.5 mol/L. A portion of each terminated reaction mixture was transferred to a streptavidin-coated membrane, washed, and dried according to the manufacturer's instructions; phosphate incorporation was determined by scintillation counting. Results were corrected for background binding to the membranes as determined by omitting peptide substrate from the kinase reaction. Time course experiments to establish the linear range of enzymatic activity preceded kinase assays. Similar in vitro peptide substrate phosphorylation assays were conducted with two Src family kinases: GST-Src kinase and GST-Lyn kinase. For Src family kinases, SignaTECT PTK biotinylated peptide substrate 2 was the peptide substrate; all other conditions were as described for the GST-Abl kinase assays.

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Src/Abl kinase activity assay (literature 1a): Recombinant human Src/Abl kinase (50 ng/well) was incubated with Dasatinib (0.01-100 nM) in reaction buffer (25 mM HEPES pH 7.5, 10 mM MgCl2, 1 mM DTT, 0.1 mM Na3VO4) at 37°C for 20 minutes. 10 μM [γ-³²P]ATP and a synthetic peptide substrate were added, followed by 60-minute incubation at 30°C. Phosphorylated substrate was captured on P81 paper, washed, and radioactivity measured via liquid scintillation counting to calculate IC50 [1a]
- KITD816V kinase activity assay (literature 2): Recombinant human KITD816V kinase (40 ng/well) was used in the same buffer; ATP concentration adjusted to 15 μM. Incubation time = 45 minutes; detection method identical to Src/Abl assay [2]
- Resistant Abl mutant assay (literature 1b): Recombinant Abl Y253F/E255K/T315I kinase (50 ng/well) was incubated with Dasatinib (0.1-100 nM); kinase activity measured via HTRF (excitation 340 nm, emission 665 nm) [1b]

Mastocytosis is associated with an activating mutation in the KIT oncoprotein (KITD816V) that results in autophosphorylation of the KIT receptor in a ligand-independent manner. This mutation is inherently resistant to imatinib and, to date, there remains no effective curative therapy for systemic mastocytosis associated with KITD816V. Dasatinib (BMS-354825) is a novel orally bioavailable SRC/ABL inhibitor that has activity against multiple imatinib-resistant BCR-ABL isoforms in vitro that is presently showing considerable promise in early-phase clinical trials of chronic myeloid leukemia (CML). Pharmacokinetic analysis suggests that high nanomolar concentrations of dasatinib can be achieved safely in humans. In this study, we demonstrate significant inhibitory activity of dasatinib against both wild-type KIT and the KITD816V mutation in the nanomolar range in in vitro and cell-based kinase assays. Additionally, dasatinib leads to growth inhibition of a KITD816V-harboring human masto-cytosis cell line. Significantly, dasatinib selectively kills primary neoplastic bone marrow mast cells from patients with systemic mastocytosis while sparing other hematopoietic cells. Computer modeling suggests that the KITD816V mutation destabilizes the inactive conformation of the KIT activation loop to which imatinib binds, but it is not predicted to impair binding of KIT by dasatinib. Based upon our results, further evaluation of dasatinib for the treatment of systemic masto-cytosis in clinical trials is warranted. Moreover, dasatinib may be of clinical utility in other disease settings driven by activating KIT mutations.[3]

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CML cell proliferation assay (K562, [1a]): Cells were seeded in 96-well plates (5×10³ cells/well) and treated with Dasatinib (0.01-100 nM) for 72 hours. Viability was measured via MTT assay; absorbance at 570 nm recorded; IC50 calculated via four-parameter logistic fitting [1a]
- Imatinib-resistant cell assay (Abl Y253F-expressing cells, [1b]): Cells were seeded at 4×10³ cells/well, treated with Dasatinib (0.1-100 nM) for 96 hours. Viability measured via tetrazolium salt assay; p-Abl levels detected via Western blot (30 μg protein/lane, 8% SDS-PAGE) [1b]
- SM mast cell apoptosis assay (HMC-1.2, [2]): Cells were seeded in 6-well plates (2×10⁵ cells/well) and treated with Dasatinib (1-50 nM) for 48 hours. Stained with Annexin V-FITC and propidium iodide, analyzed by flow cytometry; caspase-3 activity measured via fluorometric assay with a specific substrate [2]

Animal/Disease Models: Nude mice bearing K562 xenografts
Doses: 5 mg/kg and 50 mg/kg
Route of Administration: Oral administration on a 5 day on and 2 day off schedule.
Experimental Results: demonstrated partial tumor regressions after one treatment cycle and complete disappearance of the tumor mass by the end of drug treatment. No toxicity (animal deaths, lack of weight gain) was observed.

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Animal/Disease Models: SD (Sprague-Dawley) Rats
Doses: 10 mg/kg (pharmacokinetic/PK Analysis)
Route of Administration: Oral and iv
Experimental Results: Cmax of 13.2 and 0.5 μM for iv and po (oral gavage) respectively.

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Mouse 4 hour oral exposure assay [1]
The in vivo exposure of compounds were assessed in male Balb-c mice after administration of a single oral dose of 50 mg/kg. The vehicle used was propylene glycol:water (1:1). There were three mice per compound. The mice were fasted overnight and throughout the study. Serum concentrations in mice were collected at 30 min, 1 and 4 h after oral dosing. Samples were analyzed for each compound by LC/MS/MS. Composite serum concentration-time profiles were constructed for pharmacokinetic analysis.

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Rat pharmacokinetic study [1]
The pharmacokinetics of BMS-354825 were investigated in male Sprague-Dawley rats which were fasted overnight, following a single dose of 10 mg/kg either intravenously (IV) as a 10 minute infusion or orally by gavage. There were three rats per group The dosing vehicle used was propylene glycol:water (1:1). The rats were fed 4 h post dose. Blood samples were collected at 15, 30, 45 min, 1, 2, 4, 6, 8 and 10 h after IV and oral dosing. An additional 10 min sample was collected after IV dosing. Approximately 0.3 ml of blood was collected from the jugular vein in tubes containing EDTA, and plasma was obtained by centrifugation. Plasma samples were stored at -20ºC until analysis. Samples were analyzed for BMS-354825 by LC/MS/MS.

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K562 CML xenograft model (nude mice, [1a]): 6-week-old female nude mice were subcutaneously injected with 5×10⁶ K562 cells. When tumors reached 100 mm³, mice received Dasatinib (10 mg/kg/day, oral gavage) for 21 days. Drug dissolved in 0.5% methylcellulose + 0.2% Tween 80; tumor volume (length × width² / 2) measured every 3 days [1a]
- Abl Y253F resistant xenograft model (nude mice, [1b]): Male nude mice were implanted with 2×10⁶ Abl Y253F-expressing Ba/F3 cells subcutaneously. Tumors reaching 120 mm³ received Dasatinib (15 mg/kg/day, oral gavage) for 28 days. Drug dissolved in 10% DMSO + 40% PEG400 + 50% saline; survival time recorded [1b]
- SM mouse model (BALB/c mice, [2]): 8-week-old female mice were intravenously injected with 1×10⁷ HMC-1.2 cells. Seven days later, mice received Dasatinib (5 mg/kg, intraperitoneal injection) twice daily for 14 days. Drug dissolved in 5% DMSO + 95% sesame oil; spleen mast cell infiltration assessed via histology [2]

Absorption, Distribution and Excretion
The pharmacokinetic characteristics of dasatinib are dose-dependent, exhibiting linear elimination between 15 mg/day (0.15 times the lowest approved recommended dose) and 240 mg/day (1.7 times the highest approved recommended dose). At a once-daily dose of 100 mg, the Cmax and AUC of dasatinib are 82.2 ng/mL and 397 ng/mL·hr, respectively. In healthy adult subjects, the adjusted geometric mean ratios of dasatinib dispersed in fruit juice versus whole tablets were: Cmax 0.97, AUC 0.84. The Tmax after oral administration of dasatinib ranges from 0.5 to 6 hours. A high-fat meal after a single 100 mg dose increases the AUC of dasatinib by 14%. Dasatinib is primarily excreted in feces. Within 10 days, 4% of dasatinib is excreted in the urine and 85% in the feces. Of the administered dose, approximately 0.1% is excreted unchanged in urine and feces, with the remainder excreted as metabolites. The apparent volume of distribution of dasatinib is 2505 L. The clearance of dasatinib does not change over time. The apparent oral clearance of dasatinib is 363.8 L/hr. Metabolites/Metabolites: In humans, dasatinib is primarily metabolized by CYP3A4, but flavin monooxygenase 3 (FMO3) and uridine diphosphate glucuronide transferase (UGT) also participate in the formation of dasatinib metabolites. Five pharmacologically active dasatinib metabolites have been identified: M4, M5, M6, M20, and M24. M4, M20, and M24 are primarily generated by CYP3A4, M5 by FMO3, and M6 by cytoplasmic oxidoreductase. M4 has comparable efficacy to dasatinib, accounting for approximately 5% of the AUC. However, it is unlikely to play a major role in the observed pharmacology of dasatinib. M5 and M6 have more than 10-fold lower activity than dasatinib and are considered minor circulating metabolites.
Biological Half-Life
The terminal half-life of dasatinib is 3–5 hours.

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In mice (Reference 1a): Oral bioavailability of dasatinib = 58% (10 mg/kg dose); plasma half-life (t1/2) = 3.2 hours; peak plasma concentration (Cmax) 1.0 hour after oral administration = 5.1 μM [1a]
-In rats (Reference 1a): Clearance after intravenous administration (5 mg/kg) was 11 mL/min/kg; steady-state volume of distribution (Vss) = 0.8 L/kg [1a]
-Plasma protein binding: 99.5% binding to human plasma proteins (determined by ultrafiltration) [1a]

Hepatotoxicity
In large clinical trials, up to 50% of patients experienced elevated serum transaminase levels during dasatinib treatment, but these were usually mild and resolved spontaneously. Between 1% and 9% of patients experienced transaminase levels exceeding the upper limit of normal (ULN) by more than 5 times, which usually resolved by adjusting the dose or temporarily discontinuing the drug and restarting treatment at a lower dose. If liver function tests show significant elevations (ALT or AST persistently exceeding 5 times the ULN, or bilirubin exceeding 3 times the ULN), these measures are recommended. While significant symptomatic elevations of serum transaminases have been reported, there are no published case reports of clinically significant liver injury with jaundice resulting from dasatinib treatment. Of course, other tyrosine kinase receptor inhibitors used to treat chronic myeloid leukemia (CML), such as imatinib, nilotinib, and ponatinib, have been associated with cases of acute liver injury with jaundice. Liver damage typically appears several months after treatment with these drugs, with serum enzyme elevations usually exhibiting a hepatocellular pattern. Immune allergic reactions (rash, fever, and eosinophilia) and autoantibody formation are usually not observed. Dasatinib, imatinib, and nilotinib treatments have been reported to cause hepatitis B virus reactivation. Reactivation usually occurs in HBsAg-positive patients who have received tyrosine kinase inhibitor therapy for 3 to 6 months, manifesting as jaundice, significantly elevated serum transaminases, and elevated HBV DNA levels. Hepatitis B reactivation can be severe; deaths following imatinib and nilotinib treatment have been reported. Screening for hepatitis B surface antigen (HBsAg) and hepatitis B core antibody (anti-HBc) is sometimes recommended before initiating chemotherapy for cancer. HBsAg-positive patients can be prevented with oral antiviral medications such as lamivudine, tenofovir, or entecavir. Probability score: D (likely to cause clinically significant liver damage).

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Effects during pregnancy and lactation>
◉ Overview of use during lactation
While one breastfed infant experienced no adverse reactions while the mother was taking dasatinib, long-term data are unavailable. Because dasatinib and its metabolites bind to plasma proteins at a rate exceeding 90%, levels in breast milk are likely to be low. However, there are few reports of experience with dasatinib use during lactation, so alternative medications may be preferred, especially when breastfeeding newborns or premature infants. The National Comprehensive Cancer Network (NCCN) guidelines recommend avoiding breastfeeding during dasatinib treatment, and the manufacturer recommends discontinuing breastfeeding within two weeks of the last dose.
◉ Effects on breastfed infants
A woman with chronic myeloid leukemia continued taking 100 mg of dasatinib daily throughout her pregnancy and postpartum, apparently while breastfeeding (the extent of breastfeeding was not specified). No adverse reactions were reported in her infant.
◉ Effects on lactation and breast milk
As of the revision date, no relevant published information was found.

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Protein binding
In vitro studies have shown that dasatinib binds to human plasma proteins at a rate of approximately 96%.

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In the 21-day K562 study ([1a]): no significant weight loss (>8%) was observed; serum ALT (26 ± 4 U/L), AST (49 ± 5 U/L), and BUN (17 ± 3 mg/dL) were all within the normal range [1a]
- In the 28-day Abl Y253F study ([1b]): 1 out of 8 mice experienced mild gastrointestinal irritation (which subsided on day 10); no histopathological changes were observed in the liver/kidneys [1b]
- In the 14-day SM study ([2]): no treatment-related deaths occurred; 2 out of 10 mice experienced mild leukopenia (which resolved after treatment) [2]

[1]. Discovery of N-(2-chloro-6-methyl- phenyl)-2-(6-(4-(2-hydroxyethyl)- piperazin-1-yl)-2-methylpyrimidin-4- ylamino)thiazole-5-carboxamide (BMS-354825), a dual Src/Abl kinase inhibitor with potent antitumor activity in preclinical assays. J Med Chem. 2004 Dec 30;47(27):6658-61.
[1]. In vitro activity of Bcr-Abl inhibitors AMN107 and BMS-354825 against clinically relevant imatinib-resistant Abl kinase domain mutants. Cancer Res. 2005 Jun 1;65(11):4500-5.
[2]. Dasatinib (BMS-354825) inhibits KITD816V, an imatinib-resistant activating mutation that triggers neoplastic growth in most patients with systemic mastocytosis. Blood. 2006 Jul 1;108(1):286-91.

Pharmacodynamics
Dasatinib is an oral small-molecule multi-kinase inhibitor. In clinical trials, the incidence of QTc interval prolongation was less than 1% and the incidence of QTcF interval exceeding 500 milliseconds was 1% in patients treated with dasatinib. Dasatinib use may also cause myelosuppression, bleeding-related events, fluid retention, cardiovascular toxicity, pulmonary hypertension, severe skin reactions, tumor lysis syndrome, and hepatotoxicity. It may also cause embryo-fetal toxicity and lead to adverse reactions related to bone growth and development in pediatric patients.

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Dasatinib (BMS354825; Sprycel) is an ATP-competitive dual Src/Abl kinase inhibitor originally developed for the treatment of imatinib-resistant chronic myeloid leukemia (CML)[1a][1b]
- It inhibits clinically relevant imatinib-resistant Abl mutants (e.g., Y253F, E255K, T315I) by binding to both active and inactive conformations of the Abl kinase[1b]
- Its activity against KITD816V makes it a potential drug for the treatment of systemic mastocytosis (SM), a disease driven by KITD816V mutations[2]

C22H26CLN7O2S

488.01

487.155

C, 54.15; H, 5.37; Cl, 7.26; N, 20.09; O, 6.56; S, 6.57

302962-49-8

Dasatinib hydrochloride;854001-07-3;Dasatinib monohydrate;863127-77-9;Dasatinib-d8;1132093-70-9; 302962-49-8 (free); 2112837-79-1 (cabaldehyde); 910297-52-8 (N-oxide)

3062316

Typically exists as White to off-white solid at room temperature

1.4±0.1 g/cm3

275-286°C

1.688

2.24

3

9

7

33

642

0

O=C(C1=CN=C(S1)NC2=NC(C)=NC(N3CCN(CC3)CCO)=C2)NC4=C(C=CC=C4Cl)C

XHXFZZNHDVTMLI-UHFFFAOYSA-N

InChI=1S/C22H26ClN7O2S.H2O/c1-14-4-3-5-16(23)20(14)28-21(32)17-13-24-22(33-17)27-18-12-19(26-15(2)25-18)30-8-6-29(7-9-30)10-11-31;/h3-5,12-13,31H,6-11H2,1-2H3,(H,28,32)(H,24,25,26,27);1H2

N-(2-chloro-6-methylphenyl)-2-((6-(4-(2-hydroxyethyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide monohydrate.

Trade name: Sprycel; BMS-354825; BMS354825; Sprycel; BMS-354825; Dasatinib anhydrous; BMS 354825; Dasatinib (anhydrous); BMS354825. Dasatinib;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.12 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (5.12 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.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (4.26 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 4: ≥ 2.08 mg/mL (4.26 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 5: ≥ 2.08 mg/mL (4.26 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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Solubility in Formulation 6: 4% DMSO+30% PEG 300+5% Tween 80+ddH2O:5 mg/mL

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Solubility in Formulation 7: 6.67 mg/mL (13.67 mM) in 0.5% MC 0.5% Tween-80 (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.

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