Home Cytoskeletal Signaling HSP (HSP90) inhibitor Zelavespib (PU-H71) For research use only.

Zelavespib (PU-H71)

Synonyms: NSC 750424

Zelavespib (PU-H71, NSC 750424) is a potent and selective inhibitor of HSP90 with IC50 of 51 nM. Phase 1.

Zelavespib (PU-H71) Chemical Structure

Zelavespib (PU-H71) Chemical Structure

CAS No. 873436-91-0

Purity & Quality Control

Zelavespib (PU-H71) Related Products

Signaling Pathway

HSP (HSP90) Signaling Pathways

HSP (HSP90) Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
MDA-MB-468 cells Function assay Inhibitory activity against Hsp90 in human breast cancer MDA-MB-468 cell line, EC50=0.0102 μM 16392823
SKBr3 cells Growth inhibition assay Growth inhibition in human breast cancer SKBr3 cell line using SRB, IC50=0.05 μM 16392823
MCF7 cells Function assay 24 h Inhibition of Hsp90 in human MCF7 cells assessed as Her2 level after 24 hrs by Western blot, IC50=0.06 μM 18571929
MRC5 cells Cytotoxicity assay Cytotoxicity against normal lung fibroblast MRC5 cell line, IC50=1 μM 16392823
NCI-H1299 cells Function assay 12 h Reduction in oxygen consumption rate in human NCI-H1299 cells incubated for 12 hrs 25383915
NCI-H69 cells Function assay 24 h Binding affinity to HSP90 in human NCI-H69 cells after 24 hrs by fluorescence polarization assay 17603540
NCI-N417 cells Function assay 24 h Binding affinity to HSP90 in human NCI-N417 cells after 24 hrs by fluorescence polarization assay 17603540
NCI-H187 cells Function assay 24 h Binding affinity to HSP90 in human NCI-H187 cells after 24 hrs by fluorescence polarization assay 17603540
NCI-H510 cells Function assay 24 h Binding affinity to HSP90 in human NCI-H510 cells after 24 hrs by fluorescence polarization assay 17603540
SKBR3 cells Function assay 24 h Binding affinity to HSP90 in human SKBR3 cells after 24 hrs by fluorescence polarization assay 17603540
NCI-H526 cells Function assay 1 μM 24 h Binding affinity to HSP90 in human NCI-H526 cells at 1 uM after 24 hrs by fluorescence polarization assay 17603540
H69AR cells Function assay 96 h Inhibition of HSP90-mediated antiapoptotic activity in human H69AR cells assessed as induction of cell growth arrest at 10 after 96 hrs by propidium iodide staining-based flow cytometry 17603540
NCI-H526 cells Function assay 24 h Binding affinity to HSP90 in human NCI-H526 cells after 24 hrs by fluorescence polarization assay 17603540
Click to View More Cell Line Experimental Data

Biological Activity

Description Zelavespib (PU-H71, NSC 750424) is a potent and selective inhibitor of HSP90 with IC50 of 51 nM. Phase 1.
Features Purine-based, HSP90-selective inhibitor.
Targets
HSP90 [1]
51 nM
In vitro
In vitro PU-H71 (1 μM) potently suppresses the growth of triple-negative breast cancers (TNBC) cell lines MDA-MB-468, MDA-MB-231, and HCC-1806 with IC50 of 65, 140 and 87 nM, respectively. PU-H71 (1 μM) kills 80%, 65%, and 80% of the initial population of MDA-MB-468, MDA-MB-231, and HCC-1806 cells, respectively. PU-H71 (0.25-1 μM) induces a dose-dependent degradation or inactivation of tumor driving molecules, including EGFR, IGF1R, HER3, c-Kit, Raf-1and Akt. Treatment for 24 h with 1 μM PU-H71, augments the percent of cells in G2-M phase of MDA-MB-468 to 69%, mediated by reduction in CDK1 and Chk1 expression. PU-H71 induces apoptosis in TNBC in part by inactivation and downregulation of Akt and Bcl-xL. PU-H71 leads to a proteasome-mediated reduction in IRAK-1 and TBK1 levels, resulting in approximately 84% and 90% reduction in NF-κB activity in MDA-MB-231 cells treated with 0.5 and 1μM PU-H71, respectively. PU-H71 markedly contains MDA-MB-231 cell invasion, with 90% suppression at 1 μM. [1] PU-H71 (2.5 μM) generates endoplasmic reticulum (ER) stress and activated the Unfolded Protein Response (UPR) as evidenced by XBP1 mRNA splicing (2.3-fold) and up-regulation of Grp94 (3.7-fold), Grp78 (4.9-fold), and CHOP (48-fold) protein expression and ATF4 (1.8-fold) mRNA expression. PU-H71 (1 μM) induces the mitochondrial pathway of apoptosis in HeLa cells, mediated by caspase but not calpain activation. In response to PU-H71-induced ER stress, apoptosis is triggered in melanoma, cervix, colon, liver and lung cancer cells, but not in normal human fibroblasts. PU-H71 is able to induce apoptosis overcoming the resistance conferred by Bcl-2. [2] PU-H71 (30 n M) significantly reduces NOS2 activity (60% reduction) and expression in LI (1 μg/mL LPS and 5 ng/mL IFN γ)-stimulated astrocytes via inhibiting NF-κB element activation. PU-H71 displays similar effects on microglial cells as on astrocytes, with 50 nM PU-H71 needed to significantly reduce the LPS dependent nitrite release. [3]
Kinase Assay HSP90 binding assay
Measurements are performed in black 96-well microtiter plates. Cell lysates are prepared by rupturing cellular membranes by freezing at -70℃ and dissolving the cellular extract in HFB [20 mM Hepes (K), pH 7.3, 50 mM KCl, 5 mM MgCl2, 20 mM Na2MoO4, 0.01% Nonidet P-40] with added protease and phosphatase inhibitors. Saturation curves are recorded in which fluorescently labeled geldanamycin (Cy3B-GM) (3 nM) is treated with increasing amounts of cellular lysates. The amount of lysate that results in polarization (mP) readings corresponding to 90%-99% bound ligand is chosen for the competition study. Here, each 96-well plate contains 3 nM Cy3B-GM, cellular lysate (amounts as determined and normalized to total Hsp90 as determined by Western blot analysis using Hsp90 purified from HeLa cells as standard) and tested Hsp90 inhibitor in a final volume of 100 μL. The plate is left for 24 h on a shaker at 4 ℃, and the fluorescence polarization (FP) values in mP are recorded. EC50 values are determined as the competitor concentrations at which 50% of the Cy3B-GM is displaced. FP measurements are performed on an Analyst GT microplate reader.
Cell Research Cell lines Human triple-negative breast cancers cell line MDA-MB-231
Concentrations ~5μM
Incubation Time 3 days
Method

Exponentially growing MDA-MB-231 cells are seeded into black 96-well microtiter plates and incubated in medium containing either vehicle control (DMSO) or compounds for the indicated time at 37 ℃. Plates containing 3 replicate wells per assay condition are seeded at a density of 8×103 cells for each cell line in 100μL medium. After exposure of cells to the Hsp90 inhibitors, plates are equilibrated to room temperature (20-25 ℃) for approximately 30 min, and 100 μL CellTiter-Glo reagent are added to each well. Plates are mixed for 2 min on an orbital shaker and then incubated for 15 min to 2 h at room temperature. The luminescence signal in each well is measured in an Analyst GT microplate reader.
Experimental Result Images Methods Biomarkers Images PMID
Western blot AKT / c-Myc / pERK / RAF1 / EWS-FLI1 / IGF-1R / PDGFRA / c-KIT / SRC EGFR / HER3 / IGF-1R / c-Kit / Raf-1 / Akt / p90RSK / CSK / Hsp70 / Hsp90 Bcl-xl / p-PDK1 / p-AKT / cPARP LYN / SYK / BTK / AKT / MEK / ERK 24388362
Growth inhibition assay Cell viability 19416831
In Vivo
In vivo PU-H71 administered at 75 mg/kg a.d. in the MDA-MB-231 model, induces a 100% complete response, and tumors are reduced to scar tissue after 37 days of treatment, accompanied with reduction in many proliferative and anti-apoptotic molecules, namely an 80%, 95%, 99%, 80%, and 65% decrease in EGFR, HER3, Raf-1, Akt, and p-Akt, respectively. PU-H71 (75 mg/kg, 3 times per week) induces a 96% inhibition of tumor growth, accompanied by an 60% reduction in tumor cell proliferation, an 85% decline in activated Akt associated with survival and high invasive potential, and a 6-fold increase in apoptosis. [1]
Animal Research Animal Models Human triple-negative breast cancers xenografts MDA-MB-231
Dosages 75 mg/kg
Administration i.p. on an alternate day schedule
NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT05612633 Withdrawn
Accelerated Phase MPN|Blast Phase MPN
Samus Therapeutics Inc.
 June 15 2022 Phase 2
NCT03935555 Terminated
Primary Myelofibrosis (PMF)|Post-Polycythemia Vera Myelofibrosis (Post-PV MF)|Post-Essential Thrombocythemia Myelofibrosis (Post-ET MF)
Samus Therapeutics Inc.
 August 12 2019 Phase 1
NCT03373877 Terminated
Myelofibrosis|Primary Myelofibrosis|Post-polycythemia Vera Myelofibrosis|Post-essential Thrombocythemia Myelofibrosis
Samus Therapeutics Inc.
 May 24 2018 Phase 1
NCT01393509 Completed
Metastatic Solid Tumor|Lymphoma|Myeloproliferative Neoplasms (MPN)
Memorial Sloan Kettering Cancer Center
 July 6 2011 Phase 1
NCT01581541 Terminated
Solid Tumors|Lymphoma
National Cancer Institute (NCI)|National Institutes of Health Clinical Center (CC)
 April 26 2011 Phase 1

Chemical Information & Solubility

Molecular Weight 512.37 Formula

C18 H21 I N6 O2 S
CAS No. 873436-91-0 SDF --
Smiles CC(C)NCCCN1C2=NC=NC(=C2N=C1SC3=C(C=C4C(=C3)OCO4)I)N
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 100 mg/mL ( (195.17 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 100 mg/mL

Water : 34 mg/mL


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In vivo
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