Home PI3K/Akt/mTOR PI3K inhibitor - ATM/ATR inhibitor - Autophagy inhibitor - DNA-PK inhibitor - Autophagy inhibitor - PLK inhibitor Wortmannin For research use only.

Wortmannin

Synonyms: KY 12420, SL-2052, BRN 0067676, NSC 627609

Wortmannin is the first described PI3K inhibitor with IC50 of 3 nM in a cell-free assay, with little selectivity within the PI3K family. Wortmannin blocks autophagosome formation and potently inhibits DNA-PK/ATM with IC50 of 16 nM and 150 nM in cell-free assays. Wortmannin also inhibits PLK1 activity.

Wortmannin Chemical Structure

Wortmannin Chemical Structure

CAS No. 19545-26-7

Purity & Quality Control

Wortmannin Related Products

Signaling Pathway

PI3K Signaling Pathways

PI3K Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Mel-HO Apoptosis Assay 4/8 μM 24 h enhances TRAIL-induced apoptosis  24113173
A-375-TS  Apoptosis Assay 4/8 μM 24 h enhances TRAIL-induced apoptosis  24113173
A-375 Apoptosis Assay 4/8 μM 24 h enhances TRAIL-induced apoptosis  24113173
Huh7  Function Assay 3 μM 1 h reduces the virus entry into the cells 24184196
BEL/FU Function Assay 1 mM 24 h decreases protein levels of the PI3K/Akt pathway 24232099
HCT 116  Function Assay 100 nM 24 h attenuates the colonies of the tumor cells with upregulation of Akt1 24297510
HepG2 Function Assay 100 nM 24 h attenuates the colonies of the tumor cells with upregulation of Akt1 24297510
SW480  Function Assay 150nM 20 h DMSO reduces cellular accumulation of β-catenin 24324366
HEK-293 Function Assay 150nM 16 h DMSO decreases CRT activity 24324366
5637 Apoptosis Assay 10 μM 40 min reverses p21WAF1 expression, CDK expression, and cell inhibition induced by fucoidan 24333868
MG-63 Apoptosis Assay 10 µM 12 h enhances DP-induced apoptosis 24358301
H1975 Function Assay 10 μM 1 h DMSO decreases cellular phospho-AKT protein levels 24447935
H520 Function Assay 10 μM 1 h DMSO decreases cellular phospho-AKT protein levels 24447935
HepG2  Function Assay 200 nM 0.5 h attenuates FoxO phosphorylation 24535192
HL-60 Function Assay 0.1 μM 72 h blocks cell differentiation 24607273
SK-N-LO Function Assay 100 nM 0.5 h decreases the stimulant effects of on Akt phosphorylation 24654606
A549  Function Assay 10 μm  16 h DMSO modulates the IAV replication and causes retention of NP in the nucleus. 24802111
A549  Growth Inhibition Assay 3 µM  2 h suppresses Akt and GSK3β activation, S-phase arrest, cell apoptosis and caspase-3 activation 24847863
HepG2 Function Assay 100 nM 0.5 h DMSO blocks MA-induced Akt phosphorylation 24863350
MO59J Apoptosis Assay 10 μM 24 h DMSO increases the DSB level induced by or 24953561
MO59K  Apoptosis Assay 10 μM 24 h DMSO increases the DSB level induced by or 24953561
MO59J Cytotoxicity Assay 5 μM 7 d DMSO enhances the cytotoxicity of or 24953561
MO59K  Cytotoxicity Assay 5 μM 7 d DMSO enhances the cytotoxicity of or 24953561
HT-29  Growth Inhibition Assay 1.5 µM 96 h decreases cell growth which can be inhibited by KYNA 25012123
MCF7 Function Assay 100 nM 24 h eliminates E2-induced ARE-Luc activity 25172557
MDA-MB-231 Apoptosis Assay 1 μM  48 h DMSO decreases the cell survival treated with 25 μM of F1 or F2   25300932
APRE-19 Apoptosis Assay 5 μM 24 h abolishes FLZ-mediated pro-survival/anti-apoptosis activity 25329617
HUVECs Cytotoxicity Assay 100 nM 24 h attenuates the abrogative effects of calycosin on VRI-induced cytotoxicity 25450186
H1703 Growth Inhibition Assay 2.5 μM 1-4 d DMSO enhances cell growth inhibition treatment with 25490383
A459 Growth Inhibition Assay 2.5 μM 1-4 d DMSO enhances cell growth inhibition treatment with 25490383
Mel-HO-TS Apoptosis Assay 4/8 μM 24 h enhances TRAIL-induced apoptosis  24113173
MeWo Apoptosis Assay 4/8 μM 24 h enhances TRAIL-induced apoptosis  24113173
Mel-2a Apoptosis Assay 4/8 μM 24 h enhances TRAIL-induced apoptosis  24113173
MDA-MB-231 Function Assay 0–400 nM 4 h suppresses Akt phosphorylation in a dose-dependent manner 22906259
MDA-MB-231 Function Assay 400 nM 4 h decreases MMP-9 and IL-8 protein in a dose-dependent manner 22906259
Jurkat Growth Inhibition Assay 0.25-1.25 μM 24/48 h DMSO inhibits cell proliferation in both time- and dose- dependent manner 19757185
Namalwa Growth Inhibition Assay 0.25-1.25 μM 24/48 h DMSO inhibits cell proliferation in both time- and dose- dependent manner 19757185
Jurkat Apoptosis Assay 0.25-1.25 μM 24/48 h DMSO induces cell apoptosis in both time- and dose- dependent manner 19757185
Namalwa Apoptosis Assay 0.25-1.25 μM 24/48 h DMSO induces cell apoptosis in both time- and dose- dependent manner 19757185
K562 Growth Inhibition Assay 24 h IC50=25±0.14 nM 19662361
SW1990 Function Assay 0.01-1 μM 1 h inhibits HA-induced Akt phosphorylation 19469020
RT112  Growth Inhibition Assay 10 μM 24 h DMSO decreases the proportion of G2/M cells 18787832
MHG-U1 Growth Inhibition Assay 10 μM 24 h DMSO decreases the proportion of G2/M cells 18787832
SMMC-7721 Apoptosis Assay 200 nM 24 h increases CHX-induced apoptosis 17557191
SMMC-7721 Function Assay 200 nM 24 h up-regulates β1,4GT1 expression 17557191
HeLa Function Assay 100 nM  1 h alters the morphology of the transferrin recycling compartment 16890915
MRC5VI Function Assay 12.5 mM 0.5 h DMSO abolishes the Ser473/Thr308 phosphorylation of AktPKB  16227394
AT5BIVA Function Assay 12.5 mM 0.5 h DMSO abolishes the Ser473/Thr308 phosphorylation of AktPKB  16227394
M059J Function Assay 12.5 mM 0.5 h DMSO abolishes the Ser473/Thr308 phosphorylation of AktPKB  16227394
HeLa Function Assay 12.5 mM 0.5 h DMSO abolishes the Ser473/Thr308 phosphorylation of AktPKB  16227394
N2a Apoptosis Assay 0.1-10 μM 2 h induces decreased cell viability in a concentration-dependent manner 15842767
Jurkat  Kinase Assay IC50 of 24 nM 15664519
Sf9 Function assay Inhibition of human PI3Kalpha expressed in Sf9 cells by fluorescent polarization assay, IC50 = 0.012 μM. 21121631
HeLa Function assay Inhibition of AX-7503 binding to recombinant Plk3 expressed in HeLa cells by Western blot, IC50 = 0.049 μM. 17135248
HeLa Function assay Binding affinity for DNA dependent protein kinase isolated from HeLa cells; Range is 20-120, Ki = 0.12 μM. 15658870
HeLa Function assay Binding affinity for Phosphatidylinositol 3-kinase isolated from HeLa cells; Range is 20-120, Ki = 0.12 μM. 15658870
A549 Function assay Inhibition of Plk3 in human A549 cells assessed as casein substrate phosphorylation by Western blot, IC50 = 0.22 μM. 17135248
A549 Antiproliferative assay 48 hrs Antiproliferative activity against human A549 cells after 48 hrs by SRB method, IC50 = 11.4 μM. 18630894
GM00637 Function assay 1 uM Inhibition of recombinant Plk3 expressed in human GM00637 cells at 1 uM assessed as decrease in p53 serine-20 phosphorylation 17135248
MDA-MB-231 Function assay 1 to 10 uM 24 hrs Inhibition of PI3K in human MDA-MB-231 cells assessed as inhibition of AKT phosphorylation at 1 to 10 uM after 24 hrs by Western blot analysis 24828286
HeLa Function assay 100 nM 10 mins Inhibition of PI3K in human HeLa cells assessed as reduction in EGF-stimulated AKT phosphorylation at S473 at 100 nM preincubated for 10 mins followed by EGF stimulation measured after 5 mins by Western blot analysis 30380865
HeLa Function assay 100 nM 10 mins Inhibition of PI3K in human HeLa cells assessed as reduction in EGF-stimulated AKT phosphorylation at T308 at 100 nM preincubated for 10 mins followed by EGF stimulation measured after 5 mins by Western blot analysis 30380865
Click to View More Cell Line Experimental Data

Biological Activity

Description Wortmannin is the first described PI3K inhibitor with IC50 of 3 nM in a cell-free assay, with little selectivity within the PI3K family. Wortmannin blocks autophagosome formation and potently inhibits DNA-PK/ATM with IC50 of 16 nM and 150 nM in cell-free assays. Wortmannin also inhibits PLK1 activity.
Targets
PI3K [3]
(Cell-free assay)		 DNA-PK [12]
(Cell-free assay)		 ATM [12]
(Cell-free assay)		 MLCK [1]
(Cell-free assay)
3 nM 16 nM 150 nM 170 nM
In vitro
In vitro

The inhibition of MLCK by Wortmannin is not affected by calmodulin or peptide substrat, while reduced by high concentration of ATP. Wortmannin directly interacts with the catalytic domain of MLCK and leads to an irreversible loss of the enzyme activity. Wortmannin has no inhibitory to cAMP-dependent protein kinase, cGMP-dependent protein kinase, and calmodulin-dependent protein kinase II, and has little effect on protein kinase C activity. [1]

Wortmannin inhibits N-formylmethionyl-leucylphenylalanine (fMLP)-stimulated PtdInsP3 (phosphatidylinositol 3,4,5-trisphosphate) formation with IC50 of 5 nM and this inhibition is completely abolished when pretreated with 100 nM Wortmannin in human neutrophils, with increased PtdInsP2 levels and no effects on cellular PtdInsP and PtdIns contents. Wortmannin could develop oscillatory changes in F-actin content and does not inhibit fMLP-stimulated actin polymerization in neutrophils. [2]

Wortmannin irreversibly inhibits phosphatidylinositol 3-kinase (PI3-kinase) activity with binding to the 110-kDa protein (IC50 of 3 nM) and has no effect PI4-kinase in RBL-2H3 cells. Wortmannin also inhibits leukotriene release, with no effect on the activation of the tyrosine kinase Lyn. [3]

Wortmannin completely abolishes the induced hexose uptake in isolated rat adipocytes at 0.1 μM, without impairing stimulated lipolytic activity. [4]

Wortmannin suppresses induced production of nitric oxide by 50% at 500 nM in human umbilical vein endothelial cells, which is in response to IGF-1. [5]

Wortmannin suppresses DNA double strand break (DSB) repair and has no effect on DSB levels or the kinetics of single strand break (SSB) repair in Chinese hamster ovary cells at 50 μM. Wortmannin could potentiate ionizing radiation (IR)-induced cytotoxicity with no toxicity by itself. [6]

Wortmannin inhibits polo-like kinase (PLK1) activity IC50 of 24 nM in intact G2/M-arrested cells. [7]

Wortmannin increases Toll-like receptor (TLR)-mediated accumulation of IL-6 in human macrophages with EC50 of 50 nM. Meanwhile Wortmannin significantly enhances TLR-mediated inducible nitric-oxide synthase (iNOS) expression and nitrite accumulation in mouse macrphages. Wortmannin activates the nuclear factor-κB and up-regulates the cytokine mRNA production. [8]

Wortmannin also inhibits Polo-like kinase (PlK) 1 and PlK3, which play important roles in mitosis. Wortmannin treatment could lead to a reduction in phosphorylation of p53 on serine 20 induced by DNA damage. [9]

Wortmannin suppresses hyaluronan-induced Akt phosphorylation and cell motility/migration in SW1990 cells. [10]
Kinase Assay MLCK assay
MLCK activity is assayed with peptide substrate (KKRPQRATSNVFS-NH2) or myosin light chain. The peptide substrate (24 μM) is phosphorylated in a reaction mixture containing 25 mM Tris-HC1 (pH 7.5), 0.5 mg/mL bovine serum albumin, 4 mM MgCl2, 0.5 mM CaCl2, 2.6 nM calmodulin, 1.5 nM MLCK, and 400 μM ATP in a final volume of 0.25 mL. After a 10-min preincubation at 28 ºC without ATP, the reaction is started by addition of ATP at 28 ºC and terminated by the addition of 0.1 mL of 10% (v/v) acetic acid after 30 min. The mixture is analyzed by high performance liquid chromatography: column, Unisil Pack 5C18 4.6 X 150 mm; solvent, 18% (v/v) acetonitrile, 0.1% (v/v) trifluoroacetic acid in water; flow rate, 1.0 mL/min; temperature, 40 ºC; detection, absorbance at 220 nm. Percent of reaction is calculated from the ratio of peak areas of phosphorylated form to those of unphosphorylated form. Specific activity measured under the conditions described above is 0.81 μmol/min/mg. Myosin light chain (108 μg/mL) is phosphorylated in a reaction mixture containing 25 mM Tris-HC1 (pH 7.5), 0.5 mg/mL bovine serum albumin, 4 mM MgCl2, 0.5 mM CaCl2, 4.2 nM calmodulin, 0.92 nM enzyme, and 10 μM[γ-32P]ATP (100-900 cpm/pmol) in a final volume of 0.25 mL. After a 3-min preincubation at 30 ºC without ATP, the reaction is started by the addition of [γ-32P]ATP at 30 ºC and stopped by the addition of 0.125 mL of trichloroacetic acid after 5 min. The acid-precipitable materials are collected on a nitrocellulose membrane filter and washed with four 1-mL aliquots of 5% (v/v) trichloroacetic acid. The radioactivity on the filter is measured in a toluene scintillation fluid, using a Packard Tri-Carb liquid scintillation spectrometer Model 4530. The specific activity measured under the conditions is 1.23 μmol/min/mg. [1]
Cell Research Cell lines NK cells
Concentrations 1 μM
Incubation Time 1 h
Method

Primary NK cells were pretreated with wortmannin (1 μM) for 1 h, washed twice with RPMI 1640, and then treated with IL-15 (10 ng/mL) for 24 h. DMSO was used as control.
Experimental Result Images Methods Biomarkers Images PMID
Western blot p-AKT / AKT / p-GSK3β / GSK3β / Bcl-xl / Bax / Caspase-3 / Cleaved caspase-3 25344912
Immunofluorescence DNMT1 24001151
Growth inhibition assay Cell viability 25344912
In Vivo
In vivo

Wortmannin inhibits peritoneal metastasis of SW1990 in mice at 1 mg/kg, without any weight loss. [10]

Wortmannin inhibits phosphatidylinositide 3-kinase-protein kinase B (PKB)/Akt phosphorylation in both normal tissues (lung, heart and brain homogenates) and tumor tissue in mice, without mortality or acute toxicity at 0.7 mg/kg. Combination with LY188011, Wortmannin significantly increases apoptosis and inhibit tumor growth in orthotopic tumor, while both monotherapy could not. [11]
Animal Research Animal Models Human pancreatic adenocarcinoma cells PK1 are injected both s.c. and orthotopically into SCID mice.
Dosages 0.175, 0.35, and 0.7 mg/kg
Administration Injected by i.v.

Chemical Information & Solubility

Molecular Weight 428.43 Formula

C23H24O8
CAS No. 19545-26-7 SDF Download Wortmannin SDF
Smiles CC(=O)OC1CC2(C(CCC2=O)C3=C1C4(C(OC(=O)C5=COC(=C54)C3=O)COC)C)C
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 85 mg/mL ( (198.39 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble


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