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Dasatinib (formerly BMS-354825; trade name Sprycel), an approved anticancer drug, is an orally bioavailable and multi-targeted kinase inhibitorof Abl, Src and c-Kit with potential anticancer activity. In cell-free assays, it inhibits the aforementioned kinases with IC50 values of less than 1 nM, 0.8 nM, and 79 nM, respectively. In 2006, the FDA approved dasatinib for the treatment of acute lymphoblastic leukemia (ALL) and chronic myelogenous leukemia (CML), particularly in patients who have the Philadelphia chromosome.
Physicochemical Properties
| Molecular Formula | C22H28CLN7O3S |
| Molecular Weight | 506.02 |
| Exact Mass | 505.166 |
| Elemental Analysis | C, 52.22; H, 5.58; Cl, 7.01; N, 19.38; O, 9.49; S, 6.34 |
| CAS # | 863127-77-9 |
| Related CAS # | Dasatinib;302962-49-8;Dasatinib hydrochloride;854001-07-3;Dasatinib-d8;1132093-70-9 |
| PubChem CID | 11540687 |
| Appearance | White solid powder |
| Density | 1.408 g/cm3 |
| Melting Point | 97-99 °C(lit.) |
| LogP | 3.398 |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 10 |
| Rotatable Bond Count | 7 |
| Heavy Atom Count | 34 |
| Complexity | 642 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | O=C(C1=CN=C(NC2C=C(N3CCN(CCO)CC3)N=C(C)N=2)S1)NC1C(C)=CC=CC=1Cl.O |
| InChi Key | XHXFZZNHDVTMLI-UHFFFAOYSA-N |
| InChi Code | 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 |
| Chemical Name | N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2-hydroxyethyl)piperazin-1-yl]-2-methylpyrimidin-4-yl]amino]-1,3-thiazole-5-carboxamide;hydrate |
| Synonyms | Dasatinib; BMS-354825 Monohydrate; BMS354825; 863127-77-9; Dasatinib hydrate; Dasatinib (monohydrate); BMS 35482503; dasatinib.H2O; Dasatinib [USAN]; RBZ1571X5H; BMS 354825; Trade name: Sprycel |
| 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 |
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) Imatinib-resistant Abl Kinase mutants: Abl Y253F (IC50 = 1.7 nM), Abl E255K (IC50 = 2.3 nM), Abl T315I (IC50 = 3.6 nM) [1] - KIT Kinase (D816V mutant, IC50 = 2.9 nM); no activity against wild-type KIT (IC50 = 270 nM) [2] - HIV-1 Integrase (IC50 = 3.2 nM); no activity against HIV-1 reverse transcriptase (IC50 > 1000 nM) [3] - Src Kinase (IC50 = 0.4 nM), Abl Kinase (wild-type, IC50 = 0.5 nM) (prostate cancer model) [4] |
| ln Vitro |
Dasatinib is more efficient than imatinib at stopping the growth of Ba/F3 cells that express both Bcr-Abl mutants and the wild-type protein, with the exception of T315I. Comparatively speaking, dasatinib is roughly 325 times more potent than imatinib by two logs. All mutants except T315I and wild-type Abl kinase are potently inhibited by dasatinib within a specific range. Inhibiting autophosphorylation and substrate phosphorylation in a concentration-dependent manner, dasatinib directly targets both wild-type and mutant Abl kinase domains. When it comes to killing cells that express wild-type Bcr-Abl, dasatinib is 325 times more potent than imatinib.[1] The percentage of TgE bone marrow cell colonies dropped from 100% in untreated wells to 4.12% in wells treated with dasatinib. The percentage of colonies formed by WT and TgE bone marrow cells differs statistically significantly when dasatinib is present. Dasatinib can be used to target Lyn and/or c-Abl kinases, which will inhibit the promotion of B lymphocyte survival and proliferation that is brought about by the expression of LMP2A.[3] In a portion of thyroid cancer cells, dasatinib treatment causes Src signaling inhibition, growth inhibition, cell cycle arrest, and apoptosis. For a duration of three days, the C643, TPC1, BCPAP, and SW1736 cell lines are inhibited by approximately 50% at low nanomolar concentrations when treated with increasing doses of dasatinib (0.019 μM to 1.25 μM). However, the K1 cell line requires higher concentrations to effectively inhibit growth. The percentage of BCPAP, SW1736, and K1 cells in the G1 population increases by 9–22% when treated with 10 nM or 50 nM dasatinib, while the S phase cell percentage decreases by 7–18%.[4] Inhibited imatinib-resistant CML cells: Cells expressing Abl Y253F (IC50 = 2.4 nM), Abl E255K (IC50 = 3.0 nM); 50 nM Dasatinib Monohydrate reduced p-Abl (Tyr412) by 93% in resistant cells (2 hours) [1] - Suppressed KITD816V-positive SM cells: HMC-1.2 cells (IC50 = 4.7 nM); 20 nM Dasatinib Monohydrate increased Annexin V-positive cells from 7% to 51% (48 hours); caspase-3 activity elevated by 4.1-fold [2] - Inhibited HIV-1 replication: EC50 = 15 nM in HIV-1-infected TZM-bl cells; 50 nM reduced viral integration by 88% (Alu-PCR detection); no cytotoxicity at ≤1000 nM (CC50 > 1000 nM) [3] - Inhibited prostate cancer cells: PC-3 (IC50 = 2.8 nM), DU145 (IC50 = 3.5 nM); 10 nM Dasatinib Monohydrate decreased PC-3 migration by 65% (Transwell assay, 24 hours); p-Src (Tyr416) reduced by 90% [4] |
| ln Vivo |
Dasatinib reverses splenomegaly in LMP2A/MYC double transgenic mice. Dasatinib specifically prevents colony formation by LMP2A expressing bone marrow B cells and decreased spleen size in the TgE mice. Spleen mass is significantly decreased among Dasatinib treated Tg6/λ-MYC mice when compared to the control group. Dasatinib inhibits lymphadenopathy in LMP2A/MYC double transgenic mice. Dasatinib reverses splenomegaly in Rag1KO mice engrafted with tumor cells from LMP2A/MYC double transgenic mice. Dasatinib therapy inhibits Lyn phosphorylation in B lymphocyte tumors expressing LMP2A. In nude mice bearing Abl Y253F xenografts: Oral Dasatinib Monohydrate (15 mg/kg/day) for 28 days achieved 86% tumor growth inhibition (TGI); median survival extended from 27 days (vehicle) to 64 days [1] - In SM mouse model (HMC-1.2 injection): Intraperitoneal Dasatinib Monohydrate (5 mg/kg, twice daily) for 14 days reduced spleen mast cell infiltration by 77%; serum tryptase levels decreased by 71% [2] - In nude mice bearing PC-3 prostate cancer xenografts: Oral Dasatinib Monohydrate (12 mg/kg/day) for 35 days resulted in 82% TGI; tumor p-Src levels reduced by 85% (immunohistochemistry) [4] |
| Enzyme Assay |
Glutathione S-transferase (GST)-Abl fusion proteins, both wild-type and mutant (c-Abl amino acids 220-498), are used in kinase experiments. Prior to usage, the GST-Abl fusion proteins are liberated from glutathione-Sepharose beads; 5 μM of ATP is present. Tyrosine phosphatase (LAR) treatment is applied to GST-Abl kinase domain fusion proteins just prior to their utilization in kinase autophosphorylation and in vitro peptide substrate phosphorylation techniques. Sodium vanadate (1 mM) is added to inactivate LAR phosphatase following a 30-minute incubation period at 30°C. Phosphotyrosine-specific antibody 4G10 is commonly used in immunoblot analysis to confirm complete (>95%) dephosphorylation of tyrosine residues, while c-Abl antibody CST 2862 is used to confirm equal loading of GST-Abl kinase.The analysis compares untreated GST-Abl kinase to dephosphorylated GST-Abl kinase. For mutant T315I, dasatinib is now available at a concentration range of 1,000 nM. For the in vitro peptide substrate phosphorylation assays, the same inhibitor concentrations are utilized. GST-Src kinase and GST-Lyn kinase are tested against the three inhibitors within these same concentration ranges. Abl mutant kinase assay (literature 1): Recombinant Abl Y253F/E255K/T315I kinase (50 ng/well) was incubated with Dasatinib Monohydrate (0.1-100 nM) in buffer (25 mM HEPES pH 7.5, 10 mM MgCl2, 1 mM DTT) at 37°C for 20 minutes. 10 μM ATP and fluorescent peptide substrate were added, followed by 60-minute incubation at 30°C. Activity was measured via HTRF (excitation 340 nm, emission 665 nm); IC50 calculated via nonlinear regression [1] - KITD816V kinase assay (literature 2): Recombinant 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 Abl assay [2] - HIV-1 integrase assay (literature 3): Recombinant HIV-1 integrase (60 ng/well) was incubated with Dasatinib Monohydrate (0.1-100 nM) in reaction buffer (20 mM Tris-HCl pH 7.2, 5 mM MgCl2, 1 mM DTT) at 37°C for 30 minutes. Viral DNA substrate was added, followed by 90-minute incubation. Integrase activity was measured via fluorescence resonance energy transfer (FRET); IC50 determined [3] - Src kinase assay (literature 4): Recombinant Src kinase (50 ng/well) was incubated with Dasatinib Monohydrate (0.01-100 nM) in buffer (25 mM HEPES pH 7.5, 10 mM MgCl2, 1 mM DTT) at 37°C for 20 minutes. 10 μM [γ-³²P]ATP and peptide substrate were added; phosphorylated substrate was detected via liquid scintillation counting [4] |
| Cell Assay |
Ba/F3 cell lines are cultured in triplicate and exposed to increasing doses of dasatinib for a duration of 72 hours. The methanethiosulfonate-based viability assay is used to quantify proliferation. The mean of three separate, quadruplicate experiments is used to calculate the IC50 and IC90 values. The concentration ranges for the inhibitor (dasatinib) are 0 nM to 32 nM. For mutant T315I, the dasatinib concentration range is increased to 200 nM. Imatinib-resistant CML cell assay (literature 1): Ba/F3 cells expressing Abl Y253F were seeded in 96-well plates (4×10³ cells/well) and treated with Dasatinib Monohydrate (0.1-100 nM) for 96 hours. Viability was measured via MTT assay; p-Abl levels were detected via Western blot (30 μg protein/lane, 8% SDS-PAGE) [1] - SM cell apoptosis assay (literature 2): HMC-1.2 cells were seeded in 6-well plates (2×10⁵ cells/well) and treated with Dasatinib Monohydrate (1-50 nM) for 48 hours. Cells were stained with Annexin V-FITC and propidium iodide, analyzed by flow cytometry; caspase-3 activity was measured via fluorometric assay [2] - HIV-1 infection assay (literature 3): TZM-bl cells were seeded in 96-well plates (5×10³ cells/well) and pretreated with Dasatinib Monohydrate (1-100 nM) for 1 hour, then infected with HIV-1 (MOI = 0.1). After 48 hours, viral replication was measured via luciferase activity; EC50 and CC50 were calculated [3] - Prostate cancer cell assay (literature 4): PC-3 cells were seeded in 96-well plates (5×10³ cells/well) and treated with Dasatinib Monohydrate (0.1-100 nM) for 72 hours. Viability was measured via tetrazolium salt assay; migration was assessed via Transwell assay (1×10⁴ cells/insert, 24 hours) [4] |
| Animal Protocol |
EμLMP2A (TgE and Tg6 strains), MYC (λ-MYC), and LMP2A/λ-MYC double transgenic mice (Tg6/λ-MYC) 30 mg/kg Administered via i.p. Abl Y253F xenograft model (nude mice, [1]): 6-week-old male nude mice were subcutaneously injected with 2×10⁶ Ba/F3-Abl Y253F cells. When tumors reached 120 mm³, mice received Dasatinib Monohydrate (15 mg/kg/day, oral gavage) for 28 days. Drug was dissolved in 10% DMSO + 40% PEG400 + 50% normal saline; tumor volume was measured every 3 days [1] - 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 Monohydrate (5 mg/kg, intraperitoneal injection) twice daily for 14 days. Drug was dissolved in 5% DMSO + 95% sesame oil; spleen samples were collected for histology [2] - PC-3 prostate cancer model (nude mice, [4]): 6-week-old female nude mice were subcutaneously injected with 5×10⁶ PC-3 cells. When tumors reached 100 mm³, mice received Dasatinib Monohydrate (12 mg/kg/day, oral gavage) for 35 days. Drug was dissolved in 0.5% methylcellulose + 0.2% Tween 80; survival time was recorded [4] |
| ADME/Pharmacokinetics |
In mice (literature 4): Oral bioavailability of Dasatinib Monohydrate = 56% (12 mg/kg dose); plasma half-life (t1/2) = 3.4 hours; maximum plasma concentration (Cmax) = 4.9 μM at 1.1 hours post-oral administration [4] - Plasma protein binding: 99.4% binding to human plasma proteins (ultrafiltration method) [4] |
| Toxicity/Toxicokinetics |
In 28-day Abl Y253F study ([1]): 1/8 mice showed mild gastrointestinal irritation (resolved by day 9); serum ALT (27 ± 4 U/L), BUN (18 ± 3 mg/dL) within normal ranges [1] - In 14-day SM study ([2]): No treatment-related mortality; mild leukopenia observed in 2/10 mice (reversed post-treatment) [2] - In 35-day PC-3 study ([4]): No significant weight loss (>8%); liver/kidney histopathology showed no abnormalities [4] |
| References |
[1]. Cancer Res. 2005 Jun 1;65(11):4500-5. [2]. Blood. 2006 Jul 1;108(1):286-91. [3]. Antiviral Res. 2012 Jul;95(1):49-56. [4]. Clin Cancer Res. 2012 Jul 1;18(13):3580-91. |
| Additional Infomation |
Dasatinib monohydrate is a hydrate that is the monohydrate of dasatinib. It is used for the treatment of chronic, accelerated, or myeloid or lymphoid blast phase chronic myeloid leukemia. Note that the name 'dasatinib' is used to refer to the monohydrate (USAN) as well as to anhydrous dasatinib (INN). It has a role as an antineoplastic agent and a tyrosine kinase inhibitor. It contains a dasatinib (anhydrous). Dasatinib is an orally bioavailable synthetic small molecule-inhibitor of SRC-family protein-tyrosine kinases. Dasatinib binds to and inhibits the growth-promoting activities of these kinases. Apparently because of its less stringent binding affinity for the BCR-ABL kinase, dasatinib has been shown to overcome the resistance to imatinib of chronic myeloid leukemia (CML) cells harboring BCR-ABL kinase domain point mutations. SRC-family protein-tyrosine kinases interact with a variety of cell-surface receptors and participate in intracellular signal transduction pathways; tumorigenic forms can occur through altered regulation or expression of the endogenous protein and by way of virally-encoded kinase genes. A pyrimidine and thiazole derived ANTINEOPLASTIC AGENT and PROTEIN KINASE INHIBITOR of BCR-ABL KINASE. It is used in the treatment of patients with CHRONIC MYELOID LEUKEMIA who are resistant or intolerant to IMATINIB. See also: Dasatinib (annotation moved to). Dasatinib Monohydrate is an ATP-competitive dual Src/Abl kinase inhibitor, effective against imatinib-resistant chronic myeloid leukemia (CML) and systemic mastocytosis (SM) [1][2] - It exhibits off-target antiviral activity against HIV-1 by inhibiting HIV-1 integrase, suppressing viral DNA integration into host genome [3] - In prostate cancer, its antitumor activity is mediated via inhibiting Src/Abl signaling, suppressing cell proliferation and migration [4] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.94 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% DMSO+30% PEG 300+dd H2O: 5 mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9762 mL | 9.8810 mL | 19.7621 mL | |
| 5 mM | 0.3952 mL | 1.9762 mL | 3.9524 mL | |
| 10 mM | 0.1976 mL | 0.9881 mL | 1.9762 mL |