| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
AZ-628 (AZ628; AZ 628) is a novel, selective, and orally bioavailable Raf inhibitor with potential anticancer activity. In xenograft animal models, it demonstrates strong tumor growth inhibition and excellent pharmacokinetic characteristics. With respective IC50 values of 29, 34, and 105 nM, AZ-628 inhibits the Raf isoforms c-Raf1, B-RafV600E, and wild-type B-Raf. In colon and melanoma cell lines with the B-RafV600E mutation, AZ-628 potently inhibits the proliferation and causes cell cycle arrest and apoptosis.
| Targets |
c-Raf-1 (IC50 = 29 nM); B-Raf (V600E) (IC50 = 34 nM); B-Raf (IC50 = 105 nM)
AZ 628 (AZ-628; AZ628) is a pan-RAF kinase inhibitor, targeting both mutant and wild-type RAF family members. In recombinant human kinase assays, it inhibits BRAFⁿᵉᵗ/ᵛ⁶⁰⁰ᴱ (IC₅₀ = 10 nM), wild-type BRAF (BRAFʷᵗ, IC₅₀ = 30 nM), and CRAF (IC₅₀ = 25 nM) [1] - AZ 628 (AZ-628; AZ628) has minimal activity against non-RAF kinases, including MEK1 (IC₅₀ > 1 μM), EGFR (IC₅₀ > 5 μM), and PDGFRβ (IC₅₀ > 10 μM), confirming its RAF family selectivity [1] - In KRAS-mutant human colon cancer cells (HCT116), AZ 628 (AZ-628; AZ628) inhibits CRAF-mediated MEK phosphorylation with an EC₅₀ of 45 nM [2] |
|---|---|
| ln Vitro |
AZ 628 reduces the activities of preactivated B-Raf, B-RafV600E, and c-Raf-1 in in vitro kinase assays, with IC50 values of 105, 34, and 29 nM, respectively. A number of other tyrosine protein kinases, such as VEGFR2, DDR2, Lyn, Flt1, and FMS, are also prevented from activating by AZ 628. In colon and melanoma cell lines carrying the B-RafV600E mutation, AZ 628 inhibits anchorage-dependent and -independent growth, results in cell cycle arrest, and induces apoptosis[1].
AZ 628 inhibits the growth of cells that are K-RASG13D-expressing. MEK and ERK phosphorylation is reduced when RAF is inhibited with AZ 628. Viability in K-RAS mutant cells is selectively impacted by AZ 628[2]. Pan-RAF Kinase Inhibition & MAPK Pathway Blockade: In human BRAFⁿᵉᵗ/ᵛ⁶⁰⁰ᴱ-positive melanoma cells (A375), AZ 628 (AZ-628; AZ628) (0.001–10 μM) concentration-dependently reduces p-ERK levels: 0.1 μM inhibits p-ERK by 80%, and 1 μM achieves complete inhibition. It also inhibits proliferation of A375 cells with an IC₅₀ of 0.05 μM [1] - Genotype-Directed Colon Cancer Cell Activity: In human colon cancer cells, AZ 628 (AZ-628; AZ628) shows differential efficacy based on genotype: - KRAS-mutant cells (HCT116, DLD-1): IC₅₀ = 0.12–0.18 μM (proliferation inhibition), with 0.5 μM reducing p-MEK/p-ERK by 75–85% (Western blot) [2] - BRAFⁿᵉᵗ/ᵛ⁶⁰⁰ᴱ-positive cells (SW620): IC₅₀ = 0.07 μM, with 0.2 μM inhibiting p-ERK by 90% [2] - Wild-type KRAS/BRAF cells (CCD-841 CoN, normal colon epithelial): IC₅₀ > 5 μM (minimal toxicity) [2] - Apoptosis Induction in Mutant Colon Cells: In HCT116 cells (KRAS-mutant), AZ 628 (AZ-628; AZ628) (0.5–2 μM) induces apoptosis in a concentration-dependent manner: 1 μM increases Annexin V⁺ cells from 5% (vehicle) to 28%, accompanied by cleavage of caspase-3 (Western blot) [2] |
| ln Vivo |
A375 Melanoma Xenograft Model: In female nude mice bearing A375 BRAFⁿᵉᵗ/ᵛ⁶⁰⁰ᴱ xenografts, oral AZ 628 (AZ-628; AZ628) (25, 50 mg/kg/day, b.i.d.) dose-dependently inhibits tumor growth: 50 mg/kg reduces tumor volume by 75% at day 21 vs. vehicle, with no significant weight loss (<5%). Tumor p-ERK levels are reduced by 80% (immunohistochemistry) [1]
- HCT116 Colon Cancer Xenograft Model: In nude mice bearing HCT116 (KRAS-mutant) colon cancer xenografts, oral AZ 628 (AZ-628; AZ628) (30 mg/kg/day, b.i.d.) for 14 days inhibits tumor growth by 60% and reduces intratumoral p-MEK levels by 70% (Western blot of tumor lysates) [2] |
| Enzyme Assay |
AZ 628 is a pan-Raf kinase inhibitor with IC50 values for B-Raf, B-RafV600E, and c-Raf-1 of 105, 34, and 29 nM, respectively.
Recombinant BRAF/CRAF Kinase Assay: Recombinant human BRAFⁿᵉᵗ/ᵛ⁶⁰⁰ᴱ, BRAFʷᵗ, or CRAF protein (40 ng/well) was incubated in kinase buffer (50 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 1 mM DTT, 15 μM ATP) with a biotinylated MEK1 peptide (substrate, 1 μM) and various concentrations of AZ 628 (AZ-628; AZ628) (0.001–100 μM) at 30°C for 60 min. Phosphorylated substrate was detected via homogeneous time-resolved fluorescence (HTRF) using Eu-labeled anti-phospho-MEK antibody and streptavidin-allophycocyanin. Kinase activity was normalized to vehicle control, and IC₅₀ values were calculated via nonlinear regression [1] |
| Cell Assay |
In HCT-116 (K-RASG13D/+) or HKe-3 (K-RAS-/+) cell lines, cell viability was measured by Syto60 after 72 hours of treatment with AZ 628 (0.5, 1.0, and 1.5 μM), CI-1040, or BAY61-3606. Each cell line's relative cell viability is normalized to the control that received DMSO treatment[2].
Melanoma/Colon Cancer Proliferation Assay: A375/HCT116/SW620/CCD-841 CoN cells were seeded in 96-well plates (5×10³ cells/well) in DMEM + 10% FBS. After 24 h adhesion, AZ 628 (AZ-628; AZ628) (0.001–10 μM) was added, and cells were incubated for 72 h. Cell viability was measured via MTT assay (absorbance at 570 nm), and IC₅₀ values were determined using GraphPad Prism [1,2] - p-ERK/p-MEK Western Blot Assay: HCT116/A375 cells were seeded in 6-well plates (2×10⁵ cells/well) and treated with AZ 628 (AZ-628; AZ628) (0.01–2 μM) for 24 h. Cells were lysed in RIPA buffer with protease/phosphatase inhibitors, and lysates (20 μg protein/lane) were separated by SDS-PAGE. Membranes were probed with anti-p-ERK, anti-total ERK, anti-p-MEK, anti-total MEK, or anti-cleaved caspase-3 antibodies, followed by HRP-conjugated secondary antibodies. Bands were visualized via chemiluminescence [2] - Apoptosis Assay: HCT116 cells were treated with AZ 628 (AZ-628; AZ628) (0.5–2 μM) for 48 h, harvested, and stained with Annexin V-FITC and propidium iodide (PI). Apoptotic cells (Annexin V⁺/PI⁻ and Annexin V⁺/PI⁺) were quantified via flow cytometry [2] |
| Animal Protocol |
A375 Melanoma Xenograft Protocol: Female nude mice (6–7 weeks old, 18–22 g) were subcutaneously injected with A375 cells (5×10⁶ cells/mouse) in Matrigel (1:1 v/v) into the right flank. When tumors reached 100–120 mm³, mice were randomized into 3 groups (n=7/group): Vehicle (0.5% methylcellulose + 0.1% Tween 80, p.o.), AZ 628 25 mg/kg (p.o., b.i.d.), AZ 628 50 mg/kg (p.o., b.i.d.). Drugs were administered daily for 21 days. Tumor volume (V = π×L×W²/6) and body weight were measured every 3 days. At study end, tumors were excised for p-ERK immunohistochemistry [1] - HCT116 Colon Cancer Xenograft Protocol: Male nude mice (7 weeks old) were subcutaneously injected with HCT116 cells (1×10⁷ cells/mouse) in PBS. When tumors reached 150 mm³, mice were divided into 2 groups (n=6/group): Vehicle, AZ 628 30 mg/kg (p.o., b.i.d.). Treatment lasted 14 days. Tumor volume was measured every 2 days; at study end, tumors were lysed for p-MEK Western blot [2] |
| Toxicity/Toxicokinetics |
Plasma protein binding: In mouse plasma (measured by ultrafiltration), AZ 628 (AZ-628; AZ628) showed approximately 92% protein binding at concentrations of 0.01–1 μM, without concentration dependence [1]
- Acute toxicity: In nude mice treated with AZ 628 (AZ-628; AZ628) at doses up to 50 mg/kg/day, twice daily for 21 days, no death or serious toxicity was observed. Body weight remained stable, and serum ALT/AST (liver marker) and creatinine (kidney marker) were within normal ranges [1] - Normal cytotoxicity: In normal human colonic epithelial cells (CCD-841 CoN), AZ 628 (AZ-628; AZ628) (at concentrations up to 5 μM) showed extremely low cytotoxicity (cell viability reduction <10%), indicating a good therapeutic index [2] |
| References | |
| Additional Infomation |
3-(2-cyanopropyl-2-yl)-N-[4-methyl-3-[(3-methyl-4-oxo-6-quinazolinyl)amino]phenyl]benzamide is a benzamide compound. AZ 628 (AZ-628; AZ628) is an investigational pan-RAF kinase inhibitor designed to target mutant (BRAFⁿᵉᵗ/ᵛ⁶⁰⁰ᴱ) and wild-type (BRAFʷᵗ, CRAF) RAF kinases for the treatment of RAF/MAPK-driven cancers. This drug has not yet been approved for clinical use [1,2]
- Mechanism of action: Its antitumor effect is achieved by inhibiting RAF kinase activity and blocking the downstream MAPK (RAF-MEK-ERK) signaling pathway—which is continuously activated in KRAS/BRAF-mutant cancers, driving cell proliferation and survival [1,2] - Genotype selectivity: AZ 628 (AZ-628; AZ628) has preferential activity against KRAS/BRAF-mutant cancer cells and very low toxicity to normal cells, which supports its potential as a genotype-targeted therapy [2] - Research applications: This drug is widely used in preclinical studies to explore RAF family biology, activation of the MAPK pathway in mutant cancers, and to validate combination therapy strategies to overcome RAF inhibitor resistance (e.g., in combination with MEK inhibitors) [1,2] |
| Molecular Formula |
C27H25N5O2
|
|
|---|---|---|
| Molecular Weight |
451.52
|
|
| Exact Mass |
451.2
|
|
| Elemental Analysis |
C, 71.82; H, 5.58; N, 15.51; O, 7.09
|
|
| CAS # |
878739-06-1
|
|
| Related CAS # |
|
|
| PubChem CID |
11676786
|
|
| Appearance |
white solid powder
|
|
| Density |
1.2±0.1 g/cm3
|
|
| Index of Refraction |
1.638
|
|
| LogP |
2.78
|
|
| Hydrogen Bond Donor Count |
2
|
|
| Hydrogen Bond Acceptor Count |
5
|
|
| Rotatable Bond Count |
5
|
|
| Heavy Atom Count |
34
|
|
| Complexity |
845
|
|
| Defined Atom Stereocenter Count |
0
|
|
| SMILES |
N#CC(C)(C)C1C=C(C(NC2C=C(NC3C=C4C(N=CN(C)C4=O)=CC=3)C(C)=CC=2)=O)C=CC=1
|
|
| InChi Key |
ZGBGPEDJXCYQPH-UHFFFAOYSA-N
|
|
| InChi Code |
InChI=1S/C27H25N5O2/c1-17-8-9-21(31-25(33)18-6-5-7-19(12-18)27(2,3)15-28)14-24(17)30-20-10-11-23-22(13-20)26(34)32(4)16-29-23/h5-14,16,30H,1-4H3,(H,31,33)
|
|
| Chemical Name |
3-(2-cyanopropan-2-yl)-N-[4-methyl-3-[(3-methyl-4-oxoquinazolin-6-yl)amino]phenyl]benzamide
|
|
| Synonyms |
AZ-628; AZ 628; AZ628
|
|
| 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: ≥ 2.5 mg/mL (5.54 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.54 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. View More
Solubility in Formulation 3: 30% PEG400+0.5% Tween80+5% propylene glycol: 30mg/mL |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2147 mL | 11.0737 mL | 22.1474 mL | |
| 5 mM | 0.4429 mL | 2.2147 mL | 4.4295 mL | |
| 10 mM | 0.2215 mL | 1.1074 mL | 2.2147 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA