Bortezomib (PS-341)

Bortezomib (PS-341) is a potent 20S proteasome inhibitor with Ki of 0.6 nM in a cell-free assay.

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Bortezomib (PS-341) Chemical Structure

Bortezomib (PS-341) Chemical Structure
Molecular Weight: 384.24

Validation & Quality Control

Product Use Citation(141)

Customer Product Validation(14)

Quality Control & MSDS

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Product Information

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  • Research Area
  • Combination Therapy
    Combination Therapy

Product Description

Biological Activity

Description Bortezomib (PS-341) is a potent 20S proteasome inhibitor with Ki of 0.6 nM in a cell-free assay.
Targets 20S proteasome [1]
(Cell-free assay)
IC50 0.6 nM(Ki)
In vitro Bortezomib, a boronic acid dipeptide, is a highly selective, reversible inhibitor of the 26S proteasome which primarily functions in the degradation of mis-folded proteins and is essential for the regulation of the cell cycle. Exposure to Bortezomib has been shown to stabilize p21, p27, and p53, as well as the proapoptotic Bid and Bax proteins, caveolin-1, and inhibitor κB-α, which prevents activation of nuclear factor κB-induced cell survival pathways. Bortezomib also promotes the activation of the proapoptotic c-Jun-NH2 terminal kinase, as well as the endoplasmic reticulum stress response. Alteration of the levels of these cellular proteins leads to inhibition of proliferation, migration, and promotion of apoptosis of cancer cells. [2] Bortezomib is shown to penetrate into cells and inhibit proteasome-mediated intracellular proteolysis of long-lived proteins with a concentration that inhibits 50% of the proteolysis of ∼0.1 μM. The average growth inhibition of 50% value for Bortezomib across the entire panel of 60 cancer cell lines derived from multiple human tumors from the US National Cancer Institute (NCI) is 7 nM. Treatment of PC-3 cells with Bortezomib (100 nM) for 8 h results in the accumulation of cells in G2-M, with a corresponding decrease in the number of cells in G1. Bortezomib kills PC-3 cells at 24 and 48 hr with IC50 of 100 and 20 nM, respectively. Bortezomib induces nuclear condensation at 16–24 hr after treatment. Bortezomib treatment leads to PARP cleavage in a time-dependent manner with concentrations as low as 100 nM being effective at 24 hr. [1]
Cell Data
Cell LinesAssay TypeConcentrationIncubation TimeFormulationActivity Description
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NCI-H1155NEDmVXVIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=NHnTc45KSzVyPUKzNE4{OiCwTR?=
COR-L279NED5V4FIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=M2\yZ2lEPTB;MkWyMlE4KG6P
NCI-H1299NXzsb4FUT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=MliwTWM2OD1{NkGuO|Ehdk1?
EW-22MkT3S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?M4X3eGlEPTB;Mk[zMlc2KG6P
SK-MEL-2NX7XTpVIT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=MlfKTWM2OD1{OEGuPUBvVQ>?
KASUMI-1Ml7QS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?MUXJR|UxRTJ6Mz6wOUBvVQ>?
NCI-H187NW[4VJFST3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NVi5XG1CUUN3ME2yPFcvODhibl2=
NCI-H2171NYPDc2x3T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=M1j5S2lEPTB;Mki4MlkzKG6P
LNCaP-Clone-FGCMl\oS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?M3TGVWlEPTB;Mkm1MlI3KG6P
NCI-H1522MU\Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?NIDkc2RKSzVyPUOwO{4xPSCwTR?=
SCHMXLHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?M4rLc2lEPTB;M{KyMlIzKG6P
THP-1NGfhPXJIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=MlLDTWM2OD1|MkKuOkBvVQ>?
SNU-C1NHrVOnBIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=M3;XW2lEPTB;M{[yMlA6KG6P
CA46NEfaVnFIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=M{j4ZmlEPTB;M{ezMlY{KG6P
NCI-H1963NIfOd2lIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=Mlj2TWM2OD1|OE[uNVkhdk1?
DELM4PX[mdzd3e2aDDJcohq[mm2aX;uJGF{e2G7MmDOTWM2OD1|OUGuNlchdk1?
TURNUTkS3BoT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=Mom0TWM2OD1|OU[uOlEhdk1?
NCI-H226NILEcllIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=M3TXN2lEPTB;NECzMlI{KG6P
COLO-668M2jSUmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7NIjLT3hKSzVyPUSwN{42PyCwTR?=
CPC-NMnq1S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?M4XuW2lEPTB;NECzMlc4KG6P
NCI-H889NFzxPVdIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=NVTUZ4ZoUUN3ME20OlEvQTJibl2=
J-RT3-T3-5MXvHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?NWDLTFZOUUN3ME21N|IvPTdibl2=
MSTO-211HMmL4S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?M{PBWWlEPTB;NUe0MlI3KG6P
SCC-15NWPPSpJvT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NESxPHZKSzVyPU[2O{41PyCwTR?=
SUP-T1MlPZS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?NEi2b3BKSzVyPU[4Ok4xPCCwTR?=
DMS-153NIXufm5Iem:5dHigTY5pcWKrdHnvckBCe3OjeR?=NH\Y[ZdKSzVyPUe0Ok45OyCwTR?=
MS-1MVPHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?Moe1TWM2OD15NUmuOFIhdk1?
TC-YIKM3\lR2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7NHXrV3pKSzVyPUe4NU4xOSCwTR?=
RPMI-8866Mn33S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?Mn7QTWM2OD1zMEC2MlI5KM7:TR?=
KY821NVmwblI1T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm=NX31XGpIUUN3ME2xNFM3NjB2IN88US=>
P31-FUJMX;Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>?MlH2TWM2OD1zMUGyMlc2KM7:TR?=
COLO-824NE\rNGJIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=NFe1[2lKSzVyPUGyOlEvPzhizszN
U-698-MM160TGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7MXPJR|UxRTJ{NkKuNVUh|ryP
TE-441-TM{XFdmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7NGXxPHNKSzVyPUK1NlEvPyEQvF2=
IMR-5NHX6TmVIem:5dHigTY5pcWKrdHnvckBCe3OjeR?=M4PJZmlEPTB;M{SwPU43OiEQvF2=
NCI-H1838Mn\yS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl?NHW0d5BKSzVyPUSxPFYvOzJizszN

... Click to View More Cell Line Experimental Data

In vivo The anticancer effects of bortezomib as a single agent have been demonstrated in xenograft models of multiple myeloma, adult T-cell leukemia, lung, breast, prostate, pancreatic, head and neck, and colon cancer, and in melanoma. [2] Oral bortezomib 1.0 mg/ kg daily for 18 days causes tumor growth delays, as well as a decrease in the number of metastases in the Lewis lung cancer model. Bortezomib at a single dose of up to 5 mg/kg significantly decreased the surviving fraction of breast tumor cells. Bortezomib 1.0 mg/kg administrated weekly for 4 weeks reduces tumor growth by 60% in murine xenograft models of prostate cancer. 1.0 mg/kg Bortezomib administration for 4 weeks results in a 72% or 84% reduction in pancreatic cancer murine xenografts growth, as well as an increase in tumor cell apoptosis. 1.0 mg/kg Bortezomib treatment results in significant inhibition of human plasmacytoma xenograft growth, increase in tumor cells apoptosis and overall survival, and a decrease in tumor angiogenesis. [3]
Features

Protocol(Only for Reference)

Kinase Assay:

[4]

Kinetic Methods In a typical kinetic run, 2.00 mL of assay buffer (20 mM HEPES, 0.5 mM EDTA, 0.035% SDS, pH 7.8) and Suc-Leu-Leu-Val-Tyr-AMC in DMSO are added to a 3 mL fluorescence cuvette, and the cuvette is placed in the jacketed cell holder of a fluorescence spectrophotometer. Reaction temperature is maintained at 37℃ by a circulating water bath. After the reaction solution has reached thermal equilibrium (5 minutes), 1 μL−10 μL of the stock enzyme solution is added to the cuvette. Reaction progress is monitored by the increase in fluorescence emission at 440 nm (λex= 380 nm) that accompanies cleavage of AMC from peptide-AMC substrates.

Cell Assay:

[5]

Cell lines Human multiple myeloma cells line U266
Concentrations ~10 μM
Incubation Time 2 days
Method

The inhibitory effect of Bortezomib on cell growth is assessed by measuring MTT dye absorbance of the cells. Cells from 48-hour cultures are pulsed with 10 μL of 5 mg/mL MTT to each well for the last 4 hour of 48-hour cultures, followed by 100 μL of isopropanol containing 0.04 N HCl. Absorbance is measured at 570 nm using a spectrophotometer.

Animal Study:

[3]

Animal Models Human plasmacytoma xenografts RPMI 8226
Formulation Saline
Dosages 1 mg/kg
Administration i.v. twice weekly for 4 weeks, then once weekly

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

SpeciesMouseRatRabbitGuinea pigHamsterDogMonkeyBaboon
Weight (kg)0.020.151.80.40.0810312
Body Surface Area (m2)0.0070.0250.150.050.020.50.240.6
Km factor361285201220
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×  mouse Km(3)  = 11.2 mg/kg
rat Km(6)
1

References

[1] Adams J, et al. Cancer Res, 1999, 59(11), 2615-2622.

[2] Boccadoro M, et al. Cancer Cell Int, 2005, 5(1):18.

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Clinical Trial Information( data from http://clinicaltrials.gov, updated on 2015-06-27)

NCT Number Recruitment Conditions Sponsor
/Collaborators
Start Date Phases
NCT02343042 Not yet recruiting Multiple Myeloma Karyopharm Therapeutics, Inc September 2015 Phase 1|Phase 2
NCT02440464 Not yet recruiting Multiple Myeloma National Heart, Lung, and Blood Institute (NHLBI)|Blood a  ...more National Heart, Lung, and Blood Institute (NHLBI)|Blood and Marrow Transplant Clinical Trials Network|National Cancer Institute (NCI) July 2015 Phase 2
NCT02441686 Not yet recruiting Multiple Myeloma Dana-Farber Cancer Institute|Millennium Pharmaceuticals,  ...more Dana-Farber Cancer Institute|Millennium Pharmaceuticals, Inc.|Celgene Corporation June 2015 Phase 2
NCT02375555 Recruiting Multiple Myeloma Dana-Farber Cancer Institute|Bristol-Myers Squibb May 2015 Phase 2
NCT02356458 Not yet recruiting Mantle Cell Lymphoma Swiss Group for Clinical Cancer Research May 2015 Phase 1|Phase 2

view more

Chemical Information

Download Bortezomib (PS-341) SDF
Molecular Weight (MW) 384.24
Formula

C19H25BN4O4

CAS No. 179324-69-7
Storage 3 years -20℃powder
6 months-80℃in solvent
Synonyms LDP-341, MLM341
Solubility (25°C) * In vitro DMSO 76 mg/mL (197.79 mM)
Water <1 mg/mL (<1 mM)
Ethanol <1 mg/mL (<1 mM)
In vivo 0.5% methylcellulose/0.2% Tween 80 5 mg/mL
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Name B-​[(1R)​-​3-​methyl-​1-​[[(2S)​-​1-​oxo-​3-​phenyl-​2-​[(2-​pyrazinylcarbonyl)​amino]​propyl]​amino]​butyl]​-boronic acid

Customer Product Validation (14)


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Source Development 2011 138, 2903-2908. Bortezomib (PS-341) purchased from Selleck
Method Immunofluorescence
Cell Lines wild-type oocytes, Fmn2-/-oocytes
Concentrations 0.1 μM
Incubation Time 90 min
Results Endogenous Fmn2, localized in the cortex of prophase I oocytes, disappeared at NEBD, reappeared later and was enriched in the cortex opposite the spindle. Fmn2 -/-oocytes lacked cortical Fmn2 staining, suggesting that the staining is specific. Cortical localization was maintained at NEBD in wild-type oocytes treated with the proteasome inhibitor, Bortezomib, indicating that Fmn2 is degraded at NEBD.

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Source Carcinogenesis 2010 31, 1948–1955. Bortezomib (PS-341) purchased from Selleck
Method Luciferase assay
Cell Lines LNCaP-AI cells, LNCaP cells
Concentrations 20 μM
Incubation Time 24 h
Results EGF significantly stimulates the promoter activity of sPLA2-IIa gene in both LNCaP and LNCaP-AI cells (shown in LNCaP-AI cells), whereas Bortezomib and other inhibitors tested downregulated the promoter activity both at the basal level (shown in LNCaP cells) and in response to EGF stimulation (shown in LNCaP-AI cells).

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Source Carcinogenesis 2010 31, 1948–1955. Bortezomib (PS-341) purchased from Selleck
Method ELISA
Cell Lines LNCaP-AI cells
Concentrations 20 μM
Incubation Time 24 h
Results Bortezomib, Lapatinib, and LY294002 significantly inhibited sPLA2-IIa secretion, whereas Erlotinib, Gefitinib and CI-1033 had a moderate effect in LNCaP-AI cells.

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Source J Biol Chem 2010 285, 41074-41086. Bortezomib (PS-341) purchased from Selleck
Method Real Time PCR
Cell Lines HepAD38 cells
Concentrations 2-50 nM
Incubation Time 18 h
Results An inhibitory effect of proteasome inhibition by bortezomib on HBV replication can be observed in cell culture.Therefore, proteasome activation seems to be a valuable strategy of the infected cells to reduce the stress resulting from misfolded proteins.

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Source J Virol 2011 85, 2781–2792. Bortezomib (PS-341) purchased from Selleck
Method Western blot
Cell Lines MA104 cells
Concentrations 0.1-10 μM
Incubation Time 1/7 h
Results Proteasome inhibitors which have different inhibition mechanisms, such as bortezomib and lactacystin, had a strong effect on virus replication in parallel with an increase in p53 accumulation.

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Source J Virol 2011 85, 2781–2792. Bortezomib (PS-341) purchased from Selleck
Method Western blot
Cell Lines MA104 cells
Concentrations 10 μM
Incubation Time 5-10 h
Results Further kinetic experiments were performed to better define the time window within the viral replication cycle in which proteasome activity was required. For this, cells were infected for 1 h and treated with MG132 or bortezomib for 4h at different times postinfection (Fig. A). As shown in Fig. B, a more clear arrest on viral protein accumulation was observed when bortezomib or MG132 was added at relatively earlier time points. Indeed, when inhibitors were added at 5 h p.i. or later, the effect was much reduced, suggesting that once the infection was well established with a robust accumulation of viroplasms, the requirement for proteasome activity was less significant.

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Source J Virol 2011 85, 2781–2792. Bortezomib (PS-341) purchased from Selleck
Method Immunofluorescence
Cell Lines NSP5-EGFP cells
Concentrations 10 μM
Incubation Time 2-8 h
Results Both MG132 and bortezomib induced a significant arrest on the formation of viroplasms, which appeared in reduced number and with smaller size with respect to those in control cells, particularly when added at time points between 1 h and 5 h p.i.

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Source J Virol 2011 85, 2781–2792. Bortezomib (PS-341) purchased from Selleck
Method Quantification of viroplasms
Cell Lines NSP5-EGFP cells, MA104 cells
Concentrations 10 μM
Incubation Time 4 h
Results The number of viroplasms per cell in cells treated for 4 h with MG132 or bortezomib was almost the same as that found in cells fixed at the beginning of each treatment, strongly suggesting that inhibition of proteasome activity affected the assembly of new viroplasms and their growth.

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Source J Virol 2010 84, 9439–9451. Bortezomib (PS-341) purchased from Selleck
Method Proteasome activity assay, Standard plaque assay
Cell Lines A549 cells
Concentrations 10-100 nM
Incubation Time 1 h
Results As expected bortezomib treatment resulted in a block of viral replication of the avian influenzavirus strain A/FPV/Bratislava/79 (H7N7; FPV) in a concentration-dependent manner. While concentrations of 10 nM had no antiviral effect, 50 nM led to a significant titer reduction of up to 3 orders of magnitude (Fig.A). The highest concentration used (100 nM) led to a titer reduction of up to 4 orders of magnitude.These results were also confirmed in a virus growth kinetics study in infected cells that received a single dose of 50nM bortezomib. Virus titers were reduced at every time point analyzed (Fig.C).Since bortezomib is a proteasome inhibitor, we investigated whether the antiviral concentrations of bortezomib may have an inhibitory effect on the 26S proteasome in A549 cells. A concentration-dependent inhibition of the proteasome was observed in FPV-infected A549 cells at 24 h p.i. (Fig. B).

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Source J Virol 2010 84, 9439–9451. Bortezomib (PS-341) purchased from Selleck
Method Standard plaque assay, Western blot
Cell Lines A549 cells
Concentrations 50 nM
Incubation Time 1-10 h
Results A strong antiviral activity of bortezomib was observed upon addition of the compound up to 2 h p.i. At time points of 4 h p.i. and later, a dramatic decrease in the antiviral efficacy was observed. This indicates that the event in the viral life cycle that is affected by bortezomib occurs within the first 4 h. Since this correlates with the strong onset of viral gene and protein expression, we analyzed whether viral protein accumu-lation is affected by bortezomib. Indeed, we observed a strong reduction in viral matrix protein (M1) and PB1 polymerase synthesis in FPV- and PR8-infected cells at 5 h and 8 h p.i.

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Source J Virol 2010 84, 9439–9451. Bortezomib (PS-341) purchased from Selleck
Method Western blot
Cell Lines A549 cells
Concentrations 50 nM
Incubation Time 1/6/24 h
Results After a 6-h treatment with 50 nM bortezomib, activation of JNK was detected as evidenced by phosphorylation of the kinase at Thr183 and Tyr185 (lane 4). This was even enhanced upon a 24-h treatment (lane 6). Downstream substrates of JNK are the AP-1 transcription factors c-Jun and ATF-2, which are activated by JNK-mediated phosphorylation at Ser63 and Thr71, respectively. Consistent with JNK activation, we also found c-Jun and ATF-2 were phosphorylated and activated( lane 6), leading to the conclusion that, besides NF-κB, the JNK/c-Jun/ATF-2 pathway is also activated in A549 upon bortezomib treatment.

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Source PLoS One 2011 6, e23712. Bortezomib (PS-341) purchased from Selleck
Method Western blotting
Cell Lines HEK293 cells
Concentrations 50 μM
Incubation Time 4 h
Results Upon proteasome inhibition by Bortezomib an increased amount of around 5-6 fold of biotinylated molecules was detected, both in the absence and presence of US2 and US11.

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Source 2011 Mireia Vila Gasull University of Porto. Bortezomib (PS-341) purchased from Selleck
Method Western Blotting
Cell Lines KKU-M213 cells
Concentrations 0/20/40 nM
Incubation Time 0-12 h
Results We checked the effects of BTZ on the NF-κB pathway in KKU-M213 (Fig. a, b). BTZ induced IκB phosphorylation in dose- and time-dependent manners (Fig. a). Interestingly, reduced IκB was observed over time until it was barely detectable 12 h after treatment; as a result, nuclear NF-κB (p65) was markedly increased (Fig. b).

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Source Mireia Vila Gasull University of Porto. 2011;Mireia Vila Gasull . Bortezomib (PS-341) purchased from Selleck
Method PB assay
Cell Lines S2-013
Concentrations
Incubation Time 24 h
Results It is noticed more deleterious effects by free bortezomib than by BTZ CS/GA nanoparticles.

Product Use Citation (141)

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Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

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