Home Protein Tyrosine Kinase EGFR inhibitor - HER2 inhibitor - Apoptosis related activator Poziotinib (HM781-36B)

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Poziotinib (HM781-36B) EGFR inhibitor

Cat.No.S7358

Poziotinib is an irreversible pan-HER inhibitor with IC50 of 3.2 nM, 5.3 nM and 23.5 nM for HER1, HER2, and HER4, respectively. Poziotinib also induces apoptosis and G1 cell cycle arrest. Phase 2.
Poziotinib (HM781-36B) EGFR inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 491.34

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Cell Culture, Treatment & Working Concentration

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
A431 Antiproliferative assay 72 hrs Antiproliferative activity against human A431 cells expressing wild type EGFR incubated for 72 hrs by MTS assay, IC50 = 0.0009 μM. 28754471
SKBR3 Antiproliferative assay 72 hrs Antiproliferative activity against human SKBR3 cells expressing wild type HER2 incubated for 72 hrs by MTS assay, IC50 = 0.001 μM. 28754471
Ba/F3 Function assay Inhibition of human EGFR T790M/L858R mutant expressed in mouse Ba/F3 cells, IC50 = 0.0022 μM. 28754471
insect cells Function assay Inhibition of GST-tagged human EGFR catalytic domain expressed in insect cells, IC50 = 0.0032 μM. 28754471
Ba/F3 Function assay Inhibition of human EGFR T790M mutant expressed in mouse Ba/F3 cells, IC50 = 0.0042 μM. 28754471
insect cells Function assay Inhibition of GST-tagged human HER2 catalytic domain expressed in insect cells, IC50 = 0.0053 μM. 28754471
NCI-H1975 Antiproliferative assay 72 hrs Antiproliferative activity against human NCI-H1975 cells expressing EGFR T790M/L858R mutant incubated for 72 hrs by MTS assay, IC50 = 0.0057 μM. 28754471
HS27 Antiproliferative assay 72 hrs Antiproliferative activity against human HS27 cells expressing wild type HER2 incubated for 72 hrs by MTS assay, GI50 = 3.83 μM. 28754471
SKBR3 Anticancer assay 72 hrs Anticancer activity against Erb-B2 overexpressing human SKBR3 cells assessed as cell growth inhibition after 72 hrs by SRB assay, ID50 = 0.0003 μM. ChEMBL
A431 Anticancer assay 72 hrs Anticancer activity against EGFR overexpressing human A431 cells assessed as cell growth inhibition after 72 hrs by SRB assay, ID50 = 0.0004 μM. ChEMBL
Sf21 Function assay 10 mins Inhibition of N-terminal GST-tagged human EGFR (696 to end residues) expressed in baculovirus infected Sf21 cells using poly (Glu,Tyr)4:1 as substrate preincubated for 10 mins followed by substrate addition measured after 1 hr by fluorescence polarization, IC50 = 0.0013 μM. ChEMBL
NCI-H1975 Anticancer assay 48 hrs Anticancer activity against human NCI-H1975 cells assessed as cell growth inhibition after 48 hrs by celltiter one shot solution assay, IC50 = 0.0027 μM. ChEMBL
Sf21 Function assay 10 mins Inhibition of N-terminal GST-tagged recombinant human EGFR T790M mutant expressed in baculovirus infected Sf21 cells using poly (Glu,Tyr)4:1 as substrate preincubated for 10 mins followed by substrate addition measured after 1 hr by fluorescence polarizat, IC50 = 0.0044 μM. ChEMBL
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Chemical Information, Storage & Stability

Molecular Weight 491.34 Formula

C23H21Cl2FN4O3

 Storage (From the date of receipt)
CAS No. 1092364-38-9 Download SDF Storage of Stock Solutions

Synonyms HM781-36B, NOV120101 Smiles COC1=C(C=C2C(=C1)N=CN=C2NC3=C(C(=C(C=C3)Cl)Cl)F)OC4CCN(CC4)C(=O)C=C

Solubility

In vitro
Batch:

DMSO : 98 mg/mL (199.45 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molarity Calculator

Mass Concentration Volume Molecular Weight

In vivo
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Mechanism of Action

Targets/IC50/Ki
HER1 [1]
(Cell-free assay)
3.2 nM
HER2 [1]
(Cell-free assay)
5.3 nM
HER4 [1]
(Cell-free assay)
23.5 nM
In vitro
Poziotinib specifically inhibits the cell growth in HER2 amplified gastric cancer cells, and inhibits the phosphorylation of EGFR and key components of downstream signaling cascades such as STAT3, AKT and ERK. This compound also induces apoptosis and G1 cell cycle arrest by activating the mitochondrial pathway in HER2 amplified gastric cancer cells. Furthermore, it also exerts synergistic effects with chemotherapeutic agents in both HER2 amplified and HER2 non-amplified gastric cancer cells. [1]
Kinase Assay
Enzyme activity assay
To determine the IC50 values of Poziotinib for kinase inhibition, enzymes of EGFR, HER2, and HER4 are expressed as recombinant proteins in Sf9 insect cells. Enzyme selectivity screening is then performed using a tyrosine kinase assay kit. Briefly, the reactions are performed in 96 well polystyrene round-bottomed plates containing kinase buffer composed of 100 mM HEPES (pH 7.4), 25 mM MgCl2, 10 mM MnCl2 and 250 μM Na3VO4. The reactions are initiated by the addition of 100 ng/assay enzyme, 100 μM ATP, and 10 ng/mL poly(Glu, Tyr). After 1 h of incubation at room temperature, the reactions are terminated by adding 6 mM EDTA solution and then anti-phosphotyrosine antibody, PTK Green Tracer, and FP dilution buffer mixtures. The fluorescence polarization values are then measured after 30 min at room temperature using a Victor3 microplate reader. Finally, the IC50 values were calculated using the following equation: Y = bottom + (top–bottom)/(1 + 10(X-logIC50)).
In vivo
In nude mice bearing N87 human gastric cancer xenografts, Poziotinib (0.5 mg/kg p.o.) alone significantly inhibits the growth of tumors, and coadministraion of this compound and 5-FU causes more effective tumor inhibition. [1] In addition, this compound shows excellent antitumor activity in a variety of EGFR- and HER-2-dependent tumor xenograft models, including erlotinib-sensitive HCC827 NSCLC cells, erlotinib-resistant NCI-H1975 NSCLC cells, HER-2 overexpressing Calu-3 NSCLC cells, NCI-N87 gastric cancer cells, SK-Ov3 ovarian cancer cells and EGFR-overexpressing A431 epidermoid carcinoma cancer cells. [2]
References

Applications

Methods Biomarkers Images PMID
Western blot Cleaved caspase-3 / Cleaved caspase-9 / PARP / Bcl-2 pEGFR / EGFR / pHER2 / HER2 / BMX / pERK / ERK / pAKT / AKT S7358-WB1 25761479
Growth inhibition assay Cell viability S7358-viability1 25761479

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT04436562 Completed
Healthy
Spectrum Pharmaceuticals Inc
 May 29 2020 Phase 1
NCT03804515 Terminated
Solid Tumor
Spectrum Pharmaceuticals Inc
 June 25 2019 Phase 1

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