| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Focal adhesion kinase (FAK)
|
|---|---|
| ln Vitro |
1. FAK Inhibition and Cell Proliferation: In PC-3 prostate carcinoma cells, BI-853520 inhibits FAK autophosphorylation at Tyr³⁹⁷ with an IC₅₀ of 1 nM and blocks anchorage-independent colony formation with an EC₅₀ of 3 nM. Western blot analysis confirms dose-dependent reduction of p-FAK (Tyr³⁹⁷) and downstream signaling molecules (e.g., Akt, ERK) in treated cells [1][3]
2. 3D Spheroid vs. 2D Monolayer Activity: In 3D spheroid cultures of cancer cells (e.g., PC-3, MDA-MB-231), BI-853520 represses tumor cell proliferation and invasion at low doses (≤3 μM), whereas 2D monolayer cultures require ≥1000-fold higher concentrations to achieve comparable effects [1][3] 3. Breast Cancer Migration Inhibition: In highly metastatic murine breast cancer cells, BI-853520 (0.1 μM) rapidly inhibits FAK phosphorylation and reduces cell migration by 50% within 24 hours, as measured by Transwell assay [2] 4. Mesenchymal Phenotype Correlation: Sensitivity to BI-853520 in vitro is strongly associated with loss of E-cadherin expression and activation of mesenchymal markers (e.g., vimentin, N-cadherin), confirmed by immunofluorescence staining [1][3] |
| ln Vivo |
1. Adenocarcinoma Xenograft Efficacy: In nude mice bearing subcutaneous PC-3 adenocarcinoma xenografts, oral administration of BI-853520 (50 mg/kg daily) reduces tumor volume by 60–80% within 2 weeks, with complete regression in 20% of animals. Efficacy is significantly higher in tumors with mesenchymal phenotype (low E-cadherin, high miR-200c-3p), achieving median tumor growth inhibition (TGI) >100% [1][3]
2. Pharmacodynamic Target Engagement: BI-853520 rapidly penetrates tumor tissue, achieving >90% suppression of p-FAK (Tyr³⁹⁷) within 1 hour of dosing, with sustained target engagement for ≥24 hours, as shown by immunohistochemistry [1][3] 3. Breast Cancer Orthotopic Model Efficacy: In BALB/c nude mice with orthotopic MDA-MB-231 breast tumors, BI-853520 (50 mg/kg daily, oral) decreases tumor cell proliferation (Ki-67 staining) and angiogenesis (CD31⁺ vessels) by 40–60% compared to vehicle controls [2] |
| Enzyme Assay |
1. Recombinant FAK Kinase Activity Assay: Purified human FAK kinase domain is incubated with ATP (10 μM) and a fluorescent peptide substrate (sequence: KVEKIGEGTYGVVYK) in kinase buffer (50 mM Tris-HCl, pH 7.5, 10 mM MgCl₂, 1 mM DTT). BI-853520 (0.1–1000 nM) is added, and reactions are incubated at 30°C for 30 minutes. Phosphorylation is quantified using a fluorescence polarization reader, with IC₅₀ calculated as the concentration inhibiting signal by 50% [1][3]
2. Kinase Selectivity Profiling: BI-853520 is tested against a panel of 200+ recombinant kinases at 1 μM. It shows >1000-fold selectivity for FAK over other kinases (e.g., Src, Abl, VEGFR2), with inhibition rates <5% for off-target kinases [1][3] |
| Cell Assay |
1. 3D Spheroid Formation Assay: Cancer cells (5×10³ cells/well) are embedded in Matrigel (10 mg/mL) in 96-well ultra-low attachment plates and treated with BI-853520 (0.1–10 μM) for 7 days. Spheroid size is measured using brightfield microscopy (ImageJ software), and viability is assessed by Calcein-AM staining (excitation 485 nm, emission 520 nm). BI-853520 reduces spheroid diameter by ≥50% at 3 μM [1][3]
2. Transwell Migration/Invasion Assay: Transwell inserts (8-μm pores) are coated with Matrigel (50 μg/insert) for invasion assays or left uncoated for migration assays. Cells (1×10⁵ cells/insert) treated with BI-853520 (1–10 μM) are seeded in the upper chamber (serum-free medium), and the lower chamber contains 10% FBS. After 24 hours, non-migrated/invasive cells are removed, and stained cells (crystal violet) are counted under a microscope. BI-853520 inhibits migration by 60% at 10 μM in MDA-MB-231 cells [2] 3. Apoptosis Detection Assay: PC-3 cells treated with BI-853520 (5 μM) for 48 hours are stained with Annexin V-FITC and propidium iodide (PI) according to the manufacturer’s protocol. Apoptotic cells (Annexin V⁺/PI⁻ and Annexin V⁺/PI⁺) are quantified by flow cytometry, showing a 2–3-fold increase compared to vehicle controls. Caspase-3 activation is confirmed by western blot [1][3] |
| Animal Protocol |
1. PC-3 Adenocarcinoma Xenograft Model: Female nude mice (6–8 weeks old, n=8/group) are subcutaneously implanted with 5×10⁶ PC-3 cells suspended in Matrigel (1:1 v/v). When tumors reach ~100 mm³, mice receive BI-853520 formulated in 0.5% methylcellulose (w/v) via oral gavage at 50 mg/kg once daily for 21 days. Tumor volume is measured twice weekly using calipers (V = 0.5 × length × width²), and mice are euthanized on day 21. Tumors are harvested for immunohistochemistry (p-FAK, Ki-67) and western blot analysis [1][3]
2. MDA-MB-231 Breast Cancer Orthotopic Model: Female BALB/c nude mice (6–8 weeks old, n=8/group) are anesthetized, and 2×10⁶ MDA-MB-231 cells (suspended in 50 μL PBS) are injected into the fourth mammary fat pad. Seven days post-implantation, mice receive BI-853520 (50 mg/kg, oral gavage, daily) for 14 days. Primary tumor weight is measured at euthanasia, and lung metastasis is evaluated by hematoxylin-eosin (HE) staining of lung sections [2] |
| ADME/Pharmacokinetics |
1. Oral absorption and plasma pharmacokinetics: In mice, the bioavailability of oral BI-853520 (50 mg/kg) was >80%. Peak plasma concentration (Cmax) of 2-3 μM was reached 1-2 hours after administration, with a terminal half-life (t₁/₂) of 8-10 hours, supporting once-daily administration [1][3]
2. Tissue distribution and excretion: BI-853520 is widely distributed in various tissues, with a tumor/plasma concentration ratio >2:1 at steady state (day 7 of daily administration). It is mainly metabolized by hepatic CYP3A4/5, with only <5% of the dose excreted unchanged in the urine within 24 hours [1][3] |
| Toxicity/Toxicokinetics |
1. Preclinical acute and chronic toxicity: In Sprague-Dawley rats (n=6 per group), no significant changes in body weight, organ weight (liver, kidney, heart), or hematological parameters (erythrocytes, leukocytes, platelets) were observed after 28 consecutive days of oral administration of BI-853520 (50, 100, or 200 mg/kg daily). In the 200 mg/kg dose group, 10–15% of rats experienced mild transient proteinuria (<30 mg/dL), which resolved within 48 hours after drug withdrawal [1][3]. 2. Tumor tissue toxicity: In xenograft models, HE staining showed that BI-853520 (50 mg/kg daily) did not cause significant necrosis or inflammation in normal tissues (e.g., liver, kidneys) [1][3].
|
| References |
|
| Additional Infomation |
1. Mechanism of action: BI-853520 is an ATP-competitive FAK inhibitor that disrupts integrin-mediated signaling pathways, thereby reducing tumor cell adhesion, migration, and survival. It can also inhibit FAK-dependent stromal cell recruitment, thereby inhibiting the supportive role of the tumor microenvironment [1][3]
2. Sensitivity biomarkers: Preclinical studies have confirmed that the absence of E-cadherin and low expression of miR-200c-3p are predictive biomarkers for the efficacy of BI-853520, as the response rate of stromal phenotype tumors is significantly higher than that of epithelial phenotype tumors [1][3] 3. Clinical relevance to breast cancer: In breast cancer models, BI-853520 targets metastatic potential by inhibiting FAK-mediated cell migration, supporting its development in the treatment of metastatic breast cancer [2] 4. Formulation advantages: The oral formulation of BI-853520 (0.5% methylcellulose) has good stability and bioavailability, which is conducive to the translation from preclinical studies to clinical practice [1][3] |
| Molecular Formula |
C28H29CLF4N6O4
|
|---|---|
| Molecular Weight |
625.01
|
| Exact Mass |
624.187493
|
| Related CAS # |
Ifebemtinib;1227948-82-4; 2761021-80-9
|
| PubChem CID |
172227457
|
| Appearance |
Typically exists as solids at room temperature
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
12
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
43
|
| Complexity |
954
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CN1CCC(CC1)NC(=O)C2=CC(=C(C=C2F)NC3=NC=C(C(=N3)OC4=CC=CC5=C4C(=O)N(C5)C)C(F)(F)F)OC.Cl
|
| InChi Key |
BIXLZWJBRYUIKI-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C28H28F4N6O4.ClH/c1-37-9-7-16(8-10-37)34-24(39)17-11-22(41-3)20(12-19(17)29)35-27-33-13-18(28(30,31)32)25(36-27)42-21-6-4-5-15-14-38(2)26(40)23(15)21;/h4-6,11-13,16H,7-10,14H2,1-3H3,(H,34,39)(H,33,35,36);1H
|
| Chemical Name |
2-fluoro-5-methoxy-4-[[4-[(2-methyl-3-oxo-1H-isoindol-4-yl)oxy]-5-(trifluoromethyl)pyrimidin-2-yl]amino]-N-(1-methylpiperidin-4-yl)benzamide;hydrochloride
|
| Synonyms |
BI-853520 hydrochloride; Ifebemtinib (hydrochloride); SCHEMBL31008531; IN-10018 hydrochloride
|
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6000 mL | 7.9999 mL | 15.9997 mL | |
| 5 mM | 0.3200 mL | 1.6000 mL | 3.1999 mL | |
| 10 mM | 0.1600 mL | 0.8000 mL | 1.6000 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
