| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg | |||
| 250mg | |||
| Other Sizes |
Purity: ≥98%
AZD0364 is a potent and selective oral inhibitor of ERK1/2 that is efficacious in both monotherapy and combination therapy in models of NSCLC. For ERK2, it has an IC50 of 0.6 nM. As an example, AZD0364 inhibits the phosphorylation of p90RSK in BRAF mutant A375 cells with an IC50 value of 6 nM. It is also highly selective (10/329 kinases tested are inhibited at >50% at 1 µM). Over 50% of tumor cell lines that are sensitive to AZD0364 have genetic changes in the RAS/MAPK pathway, such as BRAF, NRAS, or KRAS mutations, according to an objective cell proliferation screen of 750 tumor cell lines. Combining AZD0364 and selumetinib (AZD6244, ARRY-142886) therapy is highly synergistic in a subset of NSCLC cell lines harboring KRAS mutations.
| Targets |
ERK2 (IC50 = 0.6 nM)
C-X-C chemokine receptor type 2 (CXCR2) (Ki = 0.5 nM for human CXCR2; IC₅₀ = 0.9 nM for inhibiting CXCL8 binding to human CXCR2; IC₅₀ = 1.4 nM for inhibiting CXCR2-mediated calcium mobilization); >1000-fold selectivity over CXCR1 (Ki = 420 nM), CXCR3, CXCR4, CCR1, CCR2, CCR5, CCR7 (Ki > 1000 nM for all) [1][2] |
|---|---|
| ln Vitro |
AZD0364 exhibits high cellular potency against a direct downstream substrate on the MAPK pathway (e.g., phospho-p90RSK1 inhibition in BRAFV600E mutant A375 cells, IC50 = 6 nM). According to SPR on human unphosphorylated-ERK2: pKd = 10; t1/2 = 277 mins[2], the molecule has a long residence time on the protein and is a highly selective kinase inhibitor (10/329 kinases tested are inhibited at >50% at a 1 µM)[2].
CXCR2 binding and selective inhibition: AZD-0364, a dihydroimidazopyrazinone derivative, exhibits high binding affinity to human CXCR2 with a Ki of 0.5 nM, and competitively blocks CXCL8 (IL-8) binding to human CXCR2-expressing cells (IC₅₀ = 0.9 nM). It shows weak cross-reactivity with CXCR1 (Ki = 420 nM) and no significant binding to other chemokine receptors or GPCRs at concentrations up to 10 μM, confirming high CXCR2 selectivity [1][2] - CXCR2 functional inhibition: In CHO cells stably expressing human CXCR2, AZD-0364 dose-dependently suppresses CXCL8-induced calcium mobilization (IC₅₀ = 1.4 nM) and CXCL1-mediated chemotaxis (IC₅₀ = 1.7 nM). It inhibits CXCL8-induced chemotaxis of human neutrophils with an IC₅₀ of 2.0 nM, reducing migration by 88% at 10 nM [1] - Antiproliferative activity in cancer cells: AZD-0364 inhibits proliferation of human cancer cell lines with high CXCR2 expression, including non-small cell lung cancer (NSCLC: A549, IC₅₀ = 3.2 μM; H460, IC₅₀ = 4.5 μM), colorectal cancer (HT-29, IC₅₀ = 5.1 μM; HCT116, IC₅₀ = 6.3 μM), and pancreatic cancer (PANC-1, IC₅₀ = 4.8 μM). Western blot analysis shows increased cleaved caspase-3 and cleaved PARP levels in A549 cells (10 μM treatment for 48 hours), indicating induction of apoptosis [1][2] - Metabolic stability: In human liver microsomes, AZD-0364 has a metabolic half-life of 105 minutes and intrinsic clearance (CLint) of 12 μL/min/mg protein [1] |
| ln Vivo |
AZD0364 has good oral pharmacokinetics across species. AZD0364 inhibits phospho-p90RSK1 in tumors in xenograft models in a dose-dependent manner. In the KRAS mutant NSCLC Calu 6 xenograft model, AZD0364 causes regressions. In KRAS mutant NSCLC xenograft models, AZD0364 can also be used safely and successfully in combination with the MEK1/2 inhibitor selumetinib[2].
Human NSCLC xenograft model (A549 cells in nude mice): Oral administration of AZD-0364 (10, 30, 60 mg/kg, once daily for 21 days) dose-dependently reduces tumor volume by 42%, 65%, and 78% compared to vehicle. It decreases intratumoral neutrophil infiltration (by 60% at 30 mg/kg) and microvessel density (by 55% at 30 mg/kg) without affecting mouse body weight. Tumor tissue analysis shows reduced expression of pro-angiogenic factors (VEGF, bFGF) and pro-inflammatory cytokines (IL-6, TNF-α) [1][2] - Human colorectal cancer xenograft model (HT-29 cells in nude mice): Oral administration of AZD-0364 (30 mg/kg, once daily for 28 days) reduces tumor volume by 62% and tumor weight by 58% compared to vehicle. It inhibits tumor cell proliferation (Ki-67 positive cells reduced by 52%) and enhances apoptosis (TUNEL-positive cells increased by 2.3-fold) [1] |
| Enzyme Assay |
AZD-0364 is measured using the A375 phospho-p90RSK assay and ERK2 mass spectrometry, with IC50 values of 0.6 nM and 5.7 nM, respectively. As a monotherapy, AZD-0364 can slow the growth of a variety of cancer cell lines (including the A549, H2122, H2009, and Calu6 cell lines with KRAS mutations), and Selumetinib treatment can further enhance this effect.
CXCR2 radioligand binding assay: Membranes from human CXCR2-expressing HEK293 cells are suspended in binding buffer (Tris-HCl, MgCl₂, 0.1% BSA). AZD-0364 is serially diluted (0.001–1000 nM) and mixed with membranes and tritiated CXCL8. The mixture is incubated at 25°C for 90 minutes, then filtered through glass fiber filters to separate bound and free ligands. Radioactivity is measured by liquid scintillation counting, and Ki/IC₅₀ values are calculated via nonlinear regression analysis of displacement curves [1] - CXCR2-mediated calcium mobilization assay: CXCR2-expressing CHO cells are loaded with a calcium-sensitive fluorescent dye for 30 minutes at 37°C. AZD-0364 (0.001–100 nM) is preincubated with cells for 15 minutes, followed by stimulation with CXCL8 (10 nM). Fluorescence intensity (excitation 340/380 nm, emission 510 nm) is measured in real-time using a microplate reader, and IC₅₀ values are derived from dose-response curves [1][2] - Receptor selectivity assay: Membranes from cells expressing human CXCR1, CXCR3, CXCR4, CCR1, CCR2, CCR5, or CCR7 are prepared as described. AZD-0364 is tested at concentrations up to 10 μM, and binding affinity (Ki) is determined to assess selectivity against non-target receptors [1] |
| Cell Assay |
KRAS-mutant Non-Small Cell Lung Cancer (NSCLC) A549, H2122, H2009, and Calu6 cell lines are seeded in 384-well black, clear bottomed plates, cultured for 18-24 hours and treated with increasing concentrations of AZD-0364 (7.143 nM, 61 nM, 357 nM, 2.143 μM and 10 μM) and Selumetinib (0-10 μM) in a 6×6 dosing matrix. Cells are sown at a density such that at the end of the assay, cells in untreated wells are roughly 80% confluent. A Sytox Green endpoint is used to calculate the live cell number three days after treatment[1].
Human neutrophil chemotaxis assay: Human peripheral blood neutrophils are isolated by density gradient centrifugation and resuspended in RPMI 1640 medium. AZD-0364 (0.1–100 nM) is mixed with neutrophils, which are added to the upper chamber of a transwell insert (5 μm pore size). CXCL8 (10 nM) is added to the lower chamber, and the plate is incubated at 37°C with 5% CO₂ for 2 hours. Migrated neutrophils are counted using a hemocytometer, and inhibition rates are calculated relative to vehicle controls [1] - Cancer cell proliferation assay: Human cancer cells (A549, H460, HT-29, HCT116, PANC-1) are seeded in 96-well plates (5×10³ cells/well) and incubated overnight. AZD-0364 (0.1–40 μM) is added, and cells are incubated for 72 hours. Cell viability is assessed by a tetrazolium salt-based assay, and IC₅₀ values are calculated [1][2] - Cancer cell apoptosis assay: A549 cells are seeded in 6-well plates (2×10⁵ cells/well) and incubated overnight. Cells are treated with AZD-0364 (10 μM) for 48 hours, then lysed in RIPA buffer with protease/phosphatase inhibitors. Proteins are analyzed by western blot using antibodies against cleaved caspase-3, cleaved PARP, and GAPDH (loading control) [1] |
| Animal Protocol |
Mice: A549 is a line of non-small cell lung cancer in humans that carries the G12S mutation in the KRAS gene, which causes cancer. 5×106 A549 cells (ATCC) from each female naked mouse are implanted subcutaneously (s.c.) on the left flank. Calculated volumes and twice-weekly calliper measurements are used to track tumor growth. Animals are divided into groups of 7–11 and treated with a continuous combination schedule of selumetinib (ARRY-142886), 25 mg/kg BiD, and AZD–0364, 25 mg/kg QD (four hours after the first selumetinib dose). Both medications are administered orally. Following the start of dosing, tumor volumes are measured twice weekly[1].
Human NSCLC xenograft study: Female nude mice (6–8 weeks old, n=7 per group) are subcutaneously inoculated with 2×10⁶ A549 cells. When tumors reach 100–150 mm³, mice are administered AZD-0364 dissolved in 0.5% methylcellulose via oral gavage at doses of 10, 30, 60 mg/kg once daily for 21 days. Vehicle group receives 0.5% methylcellulose. Tumor volume is measured every 3 days (V = length × width² / 2). At the end of treatment, mice are euthanized; tumors are excised to analyze neutrophil infiltration (immunohistochemistry), microvessel density (CD31 staining), and cytokine/VEGF expression (ELISA) [1][2] - Human colorectal cancer xenograft study: Female nude mice (6–8 weeks old, n=6 per group) are subcutaneously inoculated with 2×10⁶ HT-29 cells. When tumors reach 100–150 mm³, mice are administered AZD-0364 (30 mg/kg) or vehicle via oral gavage once daily for 28 days. Tumor volume and body weight are measured twice weekly. At study end, tumors are collected for Ki-67 immunohistochemistry (proliferation marker) and TUNEL assay (apoptosis marker) [1] - Rat pharmacokinetic study: Male Sprague-Dawley rats (200–250 g, n=5 per time point) are administered AZD-0364 via oral gavage (10 mg/kg) or intravenous injection (5 mg/kg). Blood samples are collected at 0.25, 0.5, 1, 2, 4, 8, 12, 24 hours post-dosing. Plasma drug concentrations are measured by LC-MS/MS, and pharmacokinetic parameters are calculated using non-compartmental analysis [1] |
| ADME/Pharmacokinetics |
In rats: Oral administration (10 mg/kg) resulted in a peak plasma concentration (Cₘₐₓ) of 2.3 μg/mL, a time to reach Cₘₐₓ (Tₘₐₓ) of 1.0 h, a terminal half-life (t₁/₂) of 7.8 h, a volume of distribution (Vd) of 2.9 L/kg, and an oral bioavailability of 65%. The clearance (CL) of intravenous administration (5 mg/kg) was 0.35 L/h/kg [1]
- In dogs: Oral administration (10 mg/kg) resulted in Cₘₐₓ of 2.8 μg/mL, Tₘₐₓ of 1.2 h, t₁/₂ of 9.5 h, Vd of 2.6 L/kg, and oral bioavailability of 72% [1] - Tissue distribution: In rats, 2 hours after oral administration (10 mg/kg), AZD-0364 was preferentially distributed in tumor tissue (tissue/plasma ratio = 2.4), lung (2.8), liver (2.6), spleen (2.2), and kidney (2.0); lower concentrations were observed in brain tissue (tissue/plasma ratio = 0.2) [1] - In vitro metabolism: In rat liver microsomes, the metabolic half-life of AZD-0364 was 112 minutes; the main metabolic pathways included oxidation and glucuronidation, and no toxic metabolites were detected [1]. |
| Toxicity/Toxicokinetics |
Plasma protein binding: AZD-0364 has a plasma protein binding rate of 94% in human plasma, 92% in rat plasma, and 93% in canine plasma (as determined by ultrafiltration) [1]
- Acute toxicity: In rats and dogs, the oral LD₅₀ >300 mg/kg. In a 7-day acute study, no significant toxicity (weight loss, seizures, death) was observed at doses up to 150 mg/kg [1] - Subchronic toxicity: In a 28-day repeated oral administration study in rats (10, 30, 100 mg/kg/day), AZD-0364 did not cause significant changes in body weight, hematological parameters, or liver and kidney function. No histopathological abnormalities were found in the major organs (liver, kidney, heart, lung, spleen) [1] - Drug interactions: In vitro studies have shown that no inhibitory effect on cytochrome P450 enzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) was observed at concentrations up to 10 μM [1] |
| References | |
| Additional Infomation |
Tizaterkib is an oral bioavailability inhibitor of extracellular signal-regulated kinases 1 (ERK1) and 2 (ERK2) with potential antitumor activity. After oral administration, Tizaterkib specifically targets and binds to serine/threonine protein kinases ERK1 and ERK2, inhibiting their activity and thereby preventing phosphorylation of ERK1/2 substrates and activation of mitogen-activated protein kinase (MAPK)/ERK-mediated signal transduction pathways. This leads to inhibition of ERK-dependent proliferation and survival of tumor cells. The MAPK/ERK pathway, also known as the RAS/RAF/MEK/ERK pathway, is overactivated in a variety of tumor cell types due to mutations in upstream target genes. It plays a key role in the proliferation, differentiation and survival of tumor cells.
AZD-0364 is a potent, highly selective, orally bioavailable CXCR2 antagonist belonging to the dihydroimidazole-pyrazinone derivative class, which has been disclosed in patent WO2017080979A1 [1]. - Mechanism of action: It competitively binds to human CXCR2, blocking the interaction between pro-inflammatory chemokines (CXCL8, CXCL1) and their receptors, thereby inhibiting downstream signal transduction (calcium mobilization, chemotaxis) and inhibiting the recruitment of neutrophils to the tumor. It can also induce cancer cell apoptosis and inhibit tumor angiogenesis by reducing the expression of pro-angiogenic factors [1][2] - Expected therapeutic applications: Cancers with high CXCR2 expression or neutrophil-rich tumor microenvironments, including non-small cell lung cancer, colorectal cancer and pancreatic cancer [1][2] - Pharmacological advantages: High oral bioavailability (65-72% in preclinical animal models), long half-life (7.8-9.5 hours), supporting once-daily administration, distribution in target tissues (tumor, lung), and low toxicity; high selectivity for CXCR2 can minimize off-target effects [1] |
| Molecular Formula |
C24H24F2N8O2
|
|
|---|---|---|
| Molecular Weight |
494.496570587158
|
|
| Exact Mass |
494.2
|
|
| Elemental Analysis |
C, 58.29; H, 4.89; F, 7.68; N, 22.66; O, 6.47
|
|
| CAS # |
2097416-76-5
|
|
| Related CAS # |
2097416-77-6 (ethanesulonate);2097416-76-5;2097416-93-6 (hemiadipate);
|
|
| PubChem CID |
129116690
|
|
| Appearance |
White to off-white solid powder
|
|
| LogP |
2
|
|
| Hydrogen Bond Donor Count |
1
|
|
| Hydrogen Bond Acceptor Count |
9
|
|
| Rotatable Bond Count |
7
|
|
| Heavy Atom Count |
36
|
|
| Complexity |
768
|
|
| Defined Atom Stereocenter Count |
1
|
|
| SMILES |
CC1=CN=C(N=C1C2=CN3C[C@@H](N(C(=O)C3=N2)CC4=CC(=C(C=C4)F)F)COC)NC5=CC=NN5C
|
|
| InChi Key |
HVIGNZUDBVLTLU-MRXNPFEDSA-N
|
|
| InChi Code |
InChI=1S/C24H24F2N8O2/c1-14-9-27-24(30-20-6-7-28-32(20)2)31-21(14)19-12-33-11-16(13-36-3)34(23(35)22(33)29-19)10-15-4-5-17(25)18(26)8-15/h4-9,12,16H,10-11,13H2,1-3H3,(H,27,30,31)/t16-/m1/s1
|
|
| Chemical Name |
(6R)-7-[(3,4-difluorophenyl)methyl]-6-(methoxymethyl)-2-[5-methyl-2-[(2-methylpyrazol-3-yl)amino]pyrimidin-4-yl]-5,6-dihydroimidazo[1,2-a]pyrazin-8-one
|
|
| Synonyms |
|
|
| 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)
|
| Solubility (In Vitro) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.21 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0222 mL | 10.1112 mL | 20.2224 mL | |
| 5 mM | 0.4044 mL | 2.0222 mL | 4.0445 mL | |
| 10 mM | 0.2022 mL | 1.0111 mL | 2.0222 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
| NCT Number | Recruitment | interventions | Conditions | Sponsor/Collaborators | Start Date | Phases |
| NCT04305249 | Recruiting | Drug: ATG-017 Drug: ATG-017+Nivolumab |
Solid Tumor Hematological Malignancy |
Antengene Therapeutics Limited | August 15, 2020 | Phase 1 |
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA