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Purity: ≥98%
Dactolisib (formerly also known as NVP-BEZ235; BEZ235; BEZ-235) is a novel, potent and dual ATP-competitive inhibitor of PI3K (phosphatidylinositol 3-kinase) and mTOR for p110α/γ/δ/β and mTOR(p70S6K) anticancer activity. With IC50 values of 4 nM, 5 nM, 7 nM, 75 nM, and 6 nM in cell-free assays, it inhibits mTOR for p110/// and mTOR(p70S6K). The first PI3K inhibitor to enter clinical trials was dactolisib (introduced in 2006), which exhibits strong antitumor efficacy against nonfunctioning pituitary adenomas. The IC50 value for it in 3T3TopBP1-ER cells is 21 nM, and it inhibits ATR. BEZ235 has both in vitro and in vivo evidence of potential anti-tumor activity. Regardless of the PI3K pathway mutation status, it stopped the growth of several cancer cell lines. It exhibited antitumor activity in xenograft mouse models and blocked PI3K signaling. It was shown through a combination study that temozolomide's effectiveness is increased.
Targets |
p110α (IC50 = 4 nM); p110α-H1047R (IC50 = 4.6 nM); p110α-E545K (IC50 = 5.7 nM); p110γ (IC50 = 5 nM); p110δ (IC50 = 7 nM); p110β (IC50 = 75 nM); mTOR (IC50 = 20.7 nM); mTORC1; mTORC2; Autophagy
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ln Vitro |
Dactolisib (BEZ235) potently inhibits PI3K in an ATP Competitive Manner. The phosphorylation levels of p70S6K, an mTOR activated kinase, were significantly lowered by the 250 nM dose of dactolisib (BEZ235). Due to the fact that the kinase domain of mTOR is highly similar to the one of class IA PI3K, Dactolisib (BEZ235) also causes a decrease in S235/S236P-RPS6 levels with an IC50 of 6.5 nM. A biochemical mTOR K-LISA assay (IC50, 20.7 nM) is used to demonstrate Dactolisib's (BEZ235) anti-mTOR activity[1].
The IC50s of Dactolisib (BEZ235) for the HCT116, DLD-1, and SW480 cell lines are 14.36.4, 9.01.5, and 12.01.6 nM, respectively[2]. |
ln Vivo |
Dactolisib (BEZ235) (45 mg/kg, p.o.) treatment induces colonic tumor regression in a GEM model for sporadic PIK3CA wild-type CRC[2]. Dactolisib (BEZ235) is given orally to MENX rats (n=2 per group) at a dose of 45 mg/kg, and the animals are sacrificed 1 or 6 hours later. When compared to rats treated with PEG, immunostains for P-AKT and P-S6 show a significant reduction of the two proteins, particularly P-S6, 6 hours after Dactolisib (BEZ235) administration. Pituitary adenomas in Dactolisib (BEZ235)-treated rats have a proteomic profile that is significantly different from tumors in placebo-treated rats six hours after treatment[3].
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Enzyme Assay |
PI3Kα, β, and δ proteins are made up of the iSH2 domain of p85 fused to the complete protein p110, with the exception of, which likewise lacks the final 20 amino acids. Full-length PI3K proteins with the first 144 amino acids deleted are generated. For easy purification, each construct is fused to a COOH-terminal His tag before being cloned into the pBlue-Bac4.5 or pVL1393 plasmids, depending on the isoform being studied. Then, using techniques suggested by the vendor for producing the appropriate recombinant baculoviruses and proteins, the various vectors are cotransfected with BaculoGold WT genomic DNA. A Kinase-Glo assay is used to assess BEZ235's ability to inhibit PI3K. The 384-well black plate is used for the kinase reaction. The PI3K proteins (10, 25, 10, and 150 nM of p110, p110, p110, and p110, respectively) are then added to each well after it has been loaded with 50 L of test items (in 90% DMSO) and 5 L of reaction buffer containing 10 g/mL PI substrate (l-phosphatidylinositol; Avanti Polar Lipids). 5 L of 1 M ATP prepared in the reaction buffer is added to begin the reaction, which is then incubated for 60 (for p110, p110, and p110) or 120 (for p110) minutes. The addition of 10 L of Kinase-Glo buffer ends the reaction. The plates are then examined for luminescence using a Synergy 2 reader. The 384-well black plate is used for the kinase reaction. The PI3K proteins (10, 25, 10, and 150 nM of p110α, p110β, p110δ, and p110γ, respectively) are then added to each well after it has been loaded with 50 μL of test items (in 90% DMSO) and 5 μL of reaction buffer containing 10 μg/mL PI substrate (l-phosphatidylinositol; Avanti Polar Lipids). 5 μL of 1 μM ATP prepared in the reaction buffer is added to begin the reaction, which is then incubated for 60 (for p110α, p110β, and p110δ) or 120 min (for p110γ) minutes. The addition of 10 μL of Kinase-Glo buffer ends the reaction. The plates are then examined for luminescence using a Synergy 2 reader.
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Cell Assay |
The human CRC cell lines, HCT116 (PIK3CA mutant; kinase domain at H1047R), DLD-1 (PIK3CA mutant; helical domain at E545K), and SW480 (PIK3CA wild-type) and isogenic DLD-1 PIK3CA mutant as well as wild-type cells are maintained in DMEM with 10% FBS and 1 × Penicillin/Streptomycin. To account for different growth kinetics, cells are plated at various initial densities (HCT116: 3 103 cells/well, DLD-1: 5.5 103 cells/well, SW480: 4.5 103 cells/well, DLD-1 PIK3CA mutant: 7 103 cells/well, and DLD-1 PIK3CA wild-type: 9 103 cells/well). BEZ235 is added to the cells at increasing concentrations after 16 hours, and the growth medium containing the drug is changed every 24 hours. According to the manufacturer's recommendations, cell viability is measured 48 hours after the start of drug treatment and 16 hours after the initial plating using the colorimetric MTS assay CellTiter 96® AQueous One Solution Cell Proliferation Assay. Cell viability following drug treatment is compared to untreated cells that were grown for 48 hours. Cells are plated with either no BEZ235 or the maximum inhibitory dose (500 nM) for 2, 6, 24, or 48 hours before being subjected to a western blot analysis.
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Animal Protocol |
Mice: An oral gavage of 45 mg/kg of BEZ235 in 10% 1-methyl-2-pyrrolidone/90% PEG 300 is administered daily for 28 days to tumor-bearing Apc CKO mice or a control vehicle alone (n = 8) for treatment. Based on evidence from the literature, the recommended dose of BEZ235 is 40–50 mg/kg body weight, which has been shown to be safe and effective in treating murine tumor models. Tumor-bearing mice are sacrificed one hour after the last dose of the drug, in accordance with pharmacokinetic studies showing that the tissue concentration of NVP-BEZ235 reaches its peak one hour after administration. Utilizing calipers to measure the width, length, and height of the colonic tumor, tumors are harvested for immunohistochemistry and western blot analysis.
Rats: MENX-affected rats used. In MENX rats, BEZ235 is tested at doses of 20, 30, and 45 mg/kg. The dose of 20 mg/kg is used for further studies because the two higher doses result in a weight loss of greater than 10% after 10 days of treatment. For MRI studies, BEZ235 (20 mg/kg) or a placebo (PEG) are given orally once daily to MENX-affected rats aged 7 to 8 months who have sizeable adenomas but are otherwise healthy. |
References |
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Molecular Formula |
C30H23N5O
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Molecular Weight |
469.5365
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Exact Mass |
469.19026
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Elemental Analysis |
C, 76.74; H, 4.94; N, 14.92; O, 3.41
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CAS # |
915019-65-7
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Related CAS # |
Dactolisib Tosylate;1028385-32-1
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Appearance |
Solid powder
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SMILES |
CC(C1=CC=C(N(C2=C3C=NC4=CC=C(C5=CC6=CC=CC=C6N=C5)C=C24)C(N3C)=O)C=C1)(C)C#N
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InChi Key |
JOGKUKXHTYWRGZ-UHFFFAOYSA-N
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InChi Code |
InChI=1S/C30H23N5O/c1-30(2,18-31)22-9-11-23(12-10-22)35-28-24-15-19(21-14-20-6-4-5-7-25(20)32-16-21)8-13-26(24)33-17-27(28)34(3)29(35)36/h4-17H,1-3H3
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Chemical Name |
2-Methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile.
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Synonyms |
BEZ 235; BEZ235; BEZ-235; Dactolisib; NVPBEZ235; NVP-BEZ 235; NVP-BEZ-235; NVP-BEZ235
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HS Tariff Code |
2934.99.9001
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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)
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Solubility (In Vitro) |
DMSO: ~0.01 mg/mL (0.02 mM)
Water: <1 mg/mL (slightly soluble or insoluble) DMF: 18 mg/mL warming (38.33 mM) |
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Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 0.52 mg/mL (1.11 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 5.2 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: ≥ 0.52 mg/mL (1.11 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 5.2 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. View More
Solubility in Formulation 3: NMP+polyethylene glycol 300 (10/90, v/v): 30mg/mL Solubility in Formulation 4: 12.5 mg/mL (26.62 mM) in 50% PEG300 50% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 5: ≥ 1.1 mg/mL (2.34 mM) (saturation unknown) in 10% 1-Methyl-2-pyrrolidinone 90% PEG300 (add these co-solvents sequentially from left to right, and one by one), clear solution. |
Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
1 mM | 2.1297 mL | 10.6487 mL | 21.2974 mL | |
5 mM | 0.4259 mL | 2.1297 mL | 4.2595 mL | |
10 mM | 0.2130 mL | 1.0649 mL | 2.1297 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.
Calculation results
Working concentration: mg/mL;
Method for preparing DMSO stock solution: mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation::Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
NCT Number | Status | Interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
NCT03373903 | Active Recruiting |
Drug: BEZ235 Drug: BEZ235 plus everolimus (RAD001) |
Respiratory Tract Infections | Restorbio Inc. | November 15, 2017 | Phase 2 |
NCT04584710 | Active Recruiting |
Drug: RTB101 Drug: Placebo |
Covid19 | Restorbio Inc. | October 13, 2020 | Phase 2 |
Effect of the dual PI3K/mTOR inhibitor NVP-BEZ235 on 3D organotypic cultures of rat primary NFPA cells. td> |
Expression of DEFB1 in human NFPAs and immortalized gonadotroph cells. Clin Cancer Res. 2015 Jul 15;21(14):3204-15. td> |
Effect of the dual PI3K/mTOR inhibitor NVP-BEZ235 in vivo. td> |
Expression ofDEFB1in human NFPAs and immortalized gonadotroph cells.Clin Cancer Res.2015 Jul 15;21(14):3204-15. td> |
Role ofDEFB1in NET cell lines.Clin Cancer Res.2015 Jul 15;21(14):3204-15. td> |
Expression ofDefb1,Tnfrsf10b, andBcl2a1in rat pituitary adenoma tissues after NVP-BEZ235 treatmentin vivo.Clin Cancer Res.2015 Jul 15;21(14):3204-15. td> |
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