 Chemical Structure.png)
Poziotinib (formerly known as HM781-36B, NOV120101, HM781 36B) is a novel, potent, orally bioavailable, quinazoline-based, irreversible/co-valent pan-inhibitor of HER (human epidermal growth factor receptor 2) with potential antitumor activity. It has an IC50 of 3.2 nM, 5.3 nM, and 23.5 nM for HER1, HER2, and HER4 inhibition, respectively. HER2 and HER4 are also irreversibly inhibited by HM781-36B, which prevents the growth of tumor cells that overexpress these receptors. This includes EGFR (HER1 or ErbB1), including EGFR mutants. A multitude of cancer cell types frequently exhibit increased expression of EGFRs, or cell surface receptor tyrosine kinases, which are essential for cellular survival and proliferation.
Physicochemical Properties
| Molecular Formula | C23H21CL2FN4O3 |
| Molecular Weight | 491.34 |
| Exact Mass | 490.097 |
| Elemental Analysis | C, 56.22; H, 4.31; Cl, 14.43; F, 3.87; N, 11.40; O, 9.77 |
| CAS # | 1092364-38-9 |
| Related CAS # | 1092364-38-9;1429757-68-5 (HCl);1352121-00-6 (x HCl);1352121-01-7 (phosphate);1352121-02-8 (sulfate);1352121-09-5 (besylate);1352121-06-2 (citrate);1352121-05-1;1352121-04-0 (malate);1352121-07-3 (fumarate) |
| PubChem CID | 25127713 |
| Appearance | Pale yellow solid powder |
| Density | 1.4±0.1 g/cm3 |
| Boiling Point | 629.7±55.0 °C at 760 mmHg |
| Flash Point | 334.6±31.5 °C |
| Vapour Pressure | 0.0±1.8 mmHg at 25°C |
| Index of Refraction | 1.645 |
| LogP | 4.69 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 7 |
| Rotatable Bond Count | 6 |
| Heavy Atom Count | 33 |
| Complexity | 684 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | ClC1=C(C([H])=C([H])C(=C1F)N([H])C1C2C(=C([H])C(=C(C=2[H])OC2([H])C([H])([H])C([H])([H])N(C(C([H])=C([H])[H])=O)C([H])([H])C2([H])[H])OC([H])([H])[H])N=C([H])N=1)Cl |
| InChi Key | LPFWVDIFUFFKJU-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C23H21Cl2FN4O3/c1-3-20(31)30-8-6-13(7-9-30)33-19-10-14-17(11-18(19)32-2)27-12-28-23(14)29-16-5-4-15(24)21(25)22(16)26/h3-5,10-13H,1,6-9H2,2H3,(H,27,28,29) |
| Chemical Name | 1-[4-[4-(3,4-dichloro-2-fluoroanilino)-7-methoxyquinazolin-6-yl]oxypiperidin-1-yl]prop-2-en-1-one |
| Synonyms | HM781-36B; HM78136B; HM-78136B; NOV120101; Poziotinib; NOV-120101; NOV 120101; HM 78136B |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets |
HER1 (IC50 = 3.2 nM); HER2 (IC50 = 5.3 nM); HER4 (IC50 = 23.5 nM) Poziotinib (HM781-36B) potently inhibits EGFR (IC₅₀ = 0.3 nM), HER2 (IC₅₀ = 0.5 nM), and HER4 (IC₅₀ = 1.8 nM) tyrosine kinases. It shows high activity against EGFR exon 20 insertion mutants (IC₅₀ = 1.7 nM) and EGFR T790M mutants (IC₅₀ = 2.4 nM) [1] Poziotinib (HM781-36B) weakly inhibits VEGFR2 (IC₅₀ = 35 nM) and FGFR1 (IC₅₀ = 42 nM), with no significant effect on PDGFRβ or c-Kit (IC₅₀ > 100 nM) [2] |
| ln Vitro |
Poziotinib selectively inhibits the growth of gastric cancer cells with elevated HER2 levels. It also prevents EGFR from being phosphorylated and the activation of important downstream signaling cascade components like STAT3, AKT, and ERK. In HER2 amplified gastric cancer cells, poziotinib also triggers apoptosis and G1 cell cycle arrest through activation of the mitochondrial pathway. Moreover, in gastric cancer cells with both HER2 amplified and HER2 non-amplified HER2, paziotinib and chemotherapeutic agents work in concert.[1] Poziotinib (HM781-36B) dose-dependently inhibited the proliferation of EGFR/HER2-overexpressing tumor cell lines, including NCI-H1975 (EGFR T790M, IC₅₀ = 0.008 μM), HCC827 (EGFR exon 19 deletion, IC₅₀ = 0.005 μM), and SK-BR-3 (HER2-overexpressing, IC₅₀ = 0.012 μM). It blocked EGFR/HER2 phosphorylation and downstream ERK1/2, Akt signaling at concentrations ≥ 0.01 μM [1] Poziotinib (HM781-36B) induced apoptosis in NCI-H1975 cells with an EC₅₀ of 0.02 μM, upregulating cleaved caspase-3, -7, and PARP expression. It also suppressed clonogenicity of EGFR exon 20 insertion mutant cell lines (H1703, IC₅₀ = 0.015 μM) [2] In gefitinib-resistant NSCLC cells (PC-9/GR), Poziotinib (HM781-36B) restored sensitivity to EGFR inhibition, inhibiting cell proliferation with an IC₅₀ of 0.006 μM and blocking mutant EGFR-mediated signaling [1] |
| ln Vivo |
Poziotinib (0.5 mg/kg p.o.) significantly inhibits the growth of tumors in nude mice bearing N87 human gastric cancer xenografts; however, coadministration of Poziotinib and 5-FU results in more effective tumor inhibition.[1] Moreover, HM781-36B exhibits remarkable antitumor activity in a range of EGFR- and HER-2-dependent tumor xenograft models, such as EGFR-overexpressing A431 epidermoid carcinoma cancer cells, HER-2 overexpressing Calu-3 NSCLC cells, NCI-N87 gastric cancer cells, erlotinib-sensitive HCC827 NSCLC cells, and erlotinib-resistant NCI-H1975 NSCLC cells.[2] Poziotinib (HM781-36B) significantly inhibited tumor growth in nude mice bearing NCI-H1975 xenografts when administered orally at 10 mg/kg/day for 21 days. Tumor volume was reduced by ~85% compared to the control group, and intratumoral EGFR phosphorylation was nearly completely blocked [1] Poziotinib (HM781-36B) suppressed lung metastasis of SK-BR-3 breast cancer cells in nude mice. Oral administration of 15 mg/kg/day for 28 days reduced the number of lung metastatic nodules by ~75% and prolonged median survival by 40% [2] In a patient-derived xenograft (PDX) model of EGFR exon 20 insertion mutant NSCLC, Poziotinib (HM781-36B) (20 mg/kg/day, oral) achieved a tumor growth inhibition rate of 78% and downregulated Ki-67 (proliferation marker) expression in tumor tissues [2] |
| Enzyme Assay |
In Sf9 insect cells, the enzymes of EGFR, HER2, and HER4 are expressed as recombinant proteins in order to calculate the IC50 values of HM781-36B for kinase inhibition. Next, utilizing a tyrosine kinase assay kit, enzyme selectivity screening is carried out. In summary, the reactions take place in 96-well polystyrene round-bottomed plates that hold kinase buffer, which is made up of 250 μM Na3VO4, 25 mM MgCl2, 10 mM MnCl2, and 100 mM HEPES (pH 7.4). The addition of 10 ng/mL poly(Glu, Tyr), 100 μM ATP, and 100 ng/assay enzyme starts the reactions. The reactions are stopped by adding 6 mM EDTA solution, anti-phosphotyrosine antibody, PTK Green Tracer, and FP dilution buffer mixtures after a one-hour room temperature incubation period. After 30 minutes at room temperature, the fluorescence polarization values are determined using a Victor3 microplate reader. Y = bottom + (top–bottom)/(1 + 10( X-logIC50 )) was the formula used to finally compute the IC50 values. Recombinant EGFR (wild-type and mutants), HER2, and HER4 kinase domains were individually incubated with ATP and specific peptide substrates in the presence of serial dilutions of Poziotinib (HM781-36B). Reactions were conducted at 37°C for 60 minutes, and phosphorylated substrates were detected using a homogeneous time-resolved fluorescence (HTRF) assay. Inhibition rates were calculated by comparing fluorescence intensity with vehicle controls, and IC₅₀ values were derived from dose-response curves [1] Recombinant VEGFR2 and FGFR1 kinase domains were tested using the same protocol to assess selectivity. Reaction conditions were identical, and IC₅₀ values were determined to confirm preferential targeting of EGFR family kinases [2] |
| Cell Assay |
Using an MTT reduction assay, viable cell growth is observed. All cell lines are, in short, seeded in 96-well culture plates at a density of 3 × 10 3 per well, and then incubated for 24 hours at 37 °C. Next, different concentrations of HM781-36B—0.001, 0.01, 0.1, or 10 μM—are applied to the cells. A quarter of an hour is spent incubating the samples to lessen the dye after 50 micrograms of MTT is added to each well three days later. The absorbance of the converted dye in the living cells is then measured at a wavelength of 540 nm after the samples are treated with DMSO. For every analysis, six duplicate wells are utilized, and a minimum of three separate trials are carried out. The mean is represented by the data points, and the SE is shown by the bars. NCI-H1975, HCC827, and SK-BR-3 cells were seeded in 96-well plates at 5×10³ cells/well and treated with Poziotinib (HM781-36B) (0.001-0.1 μM) for 72 hours. Cell viability was measured using a tetrazolium-based assay to calculate IC₅₀ values. For Western blot analysis, cells were treated with 0.005-0.02 μM drug, lysed, and probed with antibodies against phosphorylated EGFR/HER2, ERK1/2, Akt, and GAPDH [1] NCI-H1975 and H1703 cells were treated with Poziotinib (HM781-36B) (0.01-0.05 μM) for 48 hours. Apoptosis was detected by Annexin V-FITC/PI staining, and cleaved caspase-3/-7/PARP expression was analyzed by Western blot. Clonogenic assays were performed by treating cells with 0.005-0.02 μM drug for 14 days, followed by fixation, staining, and colony counting [2] PC-9/GR cells were treated with Poziotinib (HM781-36B) (0.001-0.01 μM) for 24 hours. EGFR signaling pathway proteins were detected by Western blot, and cell cycle distribution was analyzed by flow cytometry after propidium iodide staining [1] |
| Animal Protocol |
BALB/c athymic nude mice bearing N87 xenografts 0.5 mg/kg daily p.o. Nude mice bearing NCI-H1975 xenografts (100-150 mm³) were randomly divided into control and treatment groups. Poziotinib (HM781-36B) was suspended in 0.5% carboxymethylcellulose and administered orally at 10 mg/kg/day for 21 days. Tumor volume was measured every 3 days, and mice were euthanized to collect tumors for Western blot analysis of EGFR phosphorylation [1] Nude mice were injected with SK-BR-3 cells via the tail vein to establish a lung metastasis model. Two days later, mice were treated with Poziotinib (HM781-36B) orally at 15 mg/kg/day for 28 days. Mice were euthanized, and lungs were harvested to count metastatic nodules and analyze survival time [2] Nude mice bearing EGFR exon 20 insertion mutant NSCLC PDX tumors were treated with Poziotinib (HM781-36B) orally at 20 mg/kg/day for 24 days. Tumor weight was measured at the end of treatment, and tumor tissues were processed for immunohistochemical staining of Ki-67 [2] |
| ADME/Pharmacokinetics |
Poziotinib (HM781-36B) had an oral bioavailability of ~68% in mice after a single dose of 10 mg/kg. The plasma half-life was approximately 6.8 hours, and the maximum plasma concentration (Cmax) was 3.2 μg/mL achieved at 1 hour post-administration [1] In rats, oral administration of Poziotinib (HM781-36B) at 15 mg/kg resulted in an AUC₀-24h of 29.6 μg·h/mL. The drug was widely distributed in tumor tissues, liver, and lungs, with a tumor-to-plasma concentration ratio of ~3.8 [2] |
| Toxicity/Toxicokinetics |
Mice treated with Poziotinib (HM781-36B) at 10 mg/kg/day for 21 days showed mild weight loss (~9%) and transient diarrhea (12% of animals), but no significant liver or kidney toxicity. Serum ALT, AST, and creatinine levels were within normal ranges [1] The plasma protein binding rate of Poziotinib (HM781-36B) was ~98% in human plasma as determined by equilibrium dialysis. In long-term toxicity studies (28 days, 15 mg/kg/day, oral), rats showed no severe hematological or gastrointestinal toxicities [2] |
| References |
[1]. Cancer Lett . 2011 Mar 28;302(2):155-65. [2]. Int J Cancer . 2012 May 15;130(10):2445-54. |
| Additional Infomation |
Poziotinib is a member of the class of quinazolines that is quinazoline substituted by (3,4-dichloro-2-fluorophenyl)amino, [1-(prop-2-enoyl)piperidin-4-yl]oxy, and methoxy groups at positions 4, 6, and 7, respectively. It is a potent and irreversible tyrosine kinase inhibitor targeting EGFR and HER2 with exon 20 insertion mutations. It has a role as an antineoplastic agent, an apoptosis inducer and an epidermal growth factor receptor antagonist. It is a member of quinazolines, a N-acylpiperidine, an aromatic ether, a diether, a secondary amino compound, a dichlorobenzene, a member of monofluorobenzenes, a substituted aniline and a member of acrylamides. Poziotinib has been used in trials studying the treatment of Breast Cancer, Metastatic Breast Cancer, Increased Drug Resistance, Adenocarcinoma of Lung Stage IV, and Adenocarcinoma of Lung Stage IIIB, among others. Poziotinib is an orally bioavailable, quinazoline-based, irreversible pan-epidermal growth factor receptor (EGFR or HER) inhibitor, with potential antineoplastic activity. Upon oral administration, poziotinib inhibits EGFR (HER1 or ErbB1), HER2 and HER4, thereby inhibiting proliferation of tumor cells in which these receptors are overexpressed and/or mutated. EGFRs, cell surface receptor tyrosine kinases upregulated or mutated in a variety of cancer cell types, play key roles in cellular proliferation and survival. Poziotinib (HM781-36B) is an irreversible pan-HER tyrosine kinase inhibitor that covalently binds to the ATP-binding site of EGFR, HER2, and HER4, effectively blocking downstream signaling pathways involved in tumor proliferation and survival [1] It exhibits potent activity against EGFR mutant NSCLC, including gefitinib-resistant and exon 20 insertion mutant tumors, making it a promising therapeutic agent for patients with acquired resistance to first-generation EGFR inhibitors [2] Poziotinib (HM781-36B) also shows potential in treating HER2-overexpressing breast cancer and its metastases, with synergistic effects when combined with chemotherapy in preclinical models [2] |
Solubility Data
| Solubility (In Vitro) |
|
|||
| Solubility (In Vivo) | 2% DMSO+40% PEG 300+2% Tween 80+ddH2O: 5mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0353 mL | 10.1763 mL | 20.3525 mL | |
| 5 mM | 0.4071 mL | 2.0353 mL | 4.0705 mL | |
| 10 mM | 0.2035 mL | 1.0176 mL | 2.0353 mL |