Bemcentinib

Manufactured by Selleck Chemicals

Sourced in United States

Bemcentinib is a selective inhibitor of the AXL receptor tyrosine kinase. It is used in research and development applications.

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Lab products found in correlation

Shp099, by Selleck Chemicals (1 mentions) Recombinant human egf, by Thermo Fisher Scientific (1 mentions) Azd4547, by MedChemExpress (1 mentions) Cuso4, by Merck Group (1 mentions) Tris 2 carboxyethyl phosphine tcep, by Merck Group (1 mentions) Erlotinib, by LC Laboratories (1 mentions) Palbociclib, by LC Laboratories (1 mentions) Mia paca 2, by American Type Culture Collection (1 mentions) Mycoalert plus mycoplasma detection kit, by Lonza (1 mentions) Buparlisib, by MedChemExpress (1 mentions) Barasertib, by Selleck Chemicals (1 mentions) Ds ri2 camera, by Nikon (1 mentions) 24 well transwell insert, by Corning (1 mentions) Eclipse 80i microscope, by Nikon (1 mentions) Fibronectin, by Fujifilm (1 mentions) Penicillin streptomycin mixture, by Thermo Fisher Scientific (1 mentions) Chir99021, by Selleck Chemicals (1 mentions) Growth factor reduced matrigel, by Corning (1 mentions) Accutase, by Nacalai Tesque (1 mentions) A83 01, by Bio-Techne (1 mentions)

6 protocols using bemcentinib

Cell Line Authentication and Drug Screening

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All cell lines used in this study (H358, MIA PaCa-2, and H23) were obtained from the American Type Culture Collection and authenticated by the University of California (UC) Berkeley DNA Sequencing Facility using short tandem repeat DNA profiling. Cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% (v/v) fetal bovine serum, penicillin (100 U/ml), and streptomycin (100 μg/ml). All cells were grown at 37°C in 5% CO₂. All cells were tested for Mycoplasma contamination using the MycoAlert PLUS Mycoplasma Detection Kit (Lonza) before initiation of experiments. ARS-1620 was obtained by custom synthesis from Pharmaron. Erlotinib and palbociclib were obtained from LC Laboratories, AZD4547 and buparlisib were obtained from MedChem Express, and SHP099 and bemcentinib were obtained from Selleckchem. ARS-1323-alkyne was synthesized as described below. Recombinant human EGF and FGF2 (FGF-basic) were obtained from Thermo Fisher Scientific. TAMRA-N₃ and tris(benzyltriazolylmethyl)amine (TBTA) were obtained from Click Chemistry Tools. Tris(2-carboxyethyl)phosphine (TCEP) and CuSO₄ were obtained from Sigma-Aldrich.

Lou K., Steri V., Ge A.Y., Hwang Y.C., Yogodzinski C.H., Shkedi A.R., Choi A.L., Mitchell D.C., Swaney D.L., Hann B., Gordan J.D., Shokat K.M, & Gilbert L.A. (2019). KRASG12C inhibition produces a driver-limited state revealing collateral dependencies. Science signaling, 12(583), eaaw9450.

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Cell Invasion Assay with Bemcentinib

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25,000 single cells were resuspended in serum-free RPMI with or without 0.25 µM of bemcentinib (BGB324) or 0.25 µM of barasertib (both from Selleck Chem, Houston, TX). Next, cells were seeded on top of a 8 µm filter membrane within a 24-well transwell insert (Corning, New York City, NY). C/RPMI was added to the bottom of wells of 24-well Falcon TC Companion Plate (Corning, New York City, NY). After 72hrs, inserts were removed, washed with PBS and the upper surface of the membrane was scrubbed twice using cotton tipped swab to remove non-invading cells. To quantify invaded cells, the exterior bottom of each insert was fixed and stained with 25% methanol and 0.5% crystal violet solution for 5 mins. After washing of the inserts with excess distilled water and air-drying, the membranes were carefully removed using scalpel blades and mounted onto glass slides with coverslips. Slides were visualized and digital images recorded using a Nikon Eclipse 80i microscope with DS-Ri2 camera (Nikon, Tokyo, Japan) at 20x optical magnification. Cell invasion area was determined by quantifying the area with crystal violet staining using the ImageJ software.

Quah H.S., Cao E.Y., Suteja L., Li C.H., Leong H.S., Chong F.T., Gupta S., Arcinas C., Ouyang J.F., Ang V., Celhar T., Zhao Y., Tay H.C., Chan J., Takahashi T., Tan D.S., Biswas S.K., Rackham O.J, & Iyer N.G. (2023). Single cell analysis in head and neck cancer reveals potential immune evasion mechanisms during early metastasis. Nature Communications, 14, 1680.

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Establishment of an in vitro blood-brain barrier model

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All-trans RA (#0695) and A83-01 (#2939) were obtained from Tocris (Bristol, UK). FGF2 (#AF-100-18B) was obtained from PeproTech (Rocky Hill, NJ, USA). Matrigel Growth Factor Reduced (GFR) (#354230) was obtained from Corning (Corning, NY, USA). Fibronectin (#86088-83-7) was obtained from Fujifilm Wako (Osaka, Japan). Collagen type IV (#638-05921) was obtained from Nitta gelatin (Osaka, Japan). Accutase was obtained from Nacalai Tesque (Kyoto, Japan). ACE2 neutralizing antibody (#AF933, ACE2 inhibitor) was obtained from R&D Systems (Minneapolis, MN, USA). Bemcentinib (#S2841, AXL inhibitor) and CHIR99021 (#S1263) were obtained from Selleck Chemicals (Houston, TX, USA). Meplazumab (#MA5-42304, CD147 inhibitor) and penicillin–streptomycin mixture (#15140122, PS) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). EG00229 (#HY-10799, NRP1 inhibitor) was obtained from MedChemExpress (NJ, USA). Stock concentrations and solvents for chemicals were described in Table 1. All other reagents were of analytical grade and obtained from commercial sources.Table 1

Medium components used in the BBB studies

Reagent	Stock concentration	Solvent
All-trans RA	40 mM	DMSO
A83-01	10 mM	DMSO
FGF2	100 µg/mL	water
CHIR99021	30 mM	DMSO
ACE2 Ab	200 µg/mL	PBS
Bemcentinib	10 mM	DMSO
Meplazumab	930 µg/mL	PBS
EG00229	100 mM	DMSO

Yamada S., Hashita T., Yanagida S., Sato H., Yasuhiko Y., Okabe K., Noda T., Nishida M., Matsunaga T, & Kanda Y. (2024). SARS-CoV-2 causes dysfunction in human iPSC-derived brain microvascular endothelial cells potentially by modulating the Wnt signaling pathway. Fluids and Barriers of the CNS, 21, 32.

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AXL Knockout Cell Lines for Drug Evaluation

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Cells were provided by the Moffitt Lung Cancer Center of Excellence Cell Line Core. The H1299 AXL KO and PC9 AXL KO cells were developed in the Meyer lab at UCLA. Cells were confirmed to be free of mycoplasma (PlasmoTest, Invivogen) and authenticated by short tandem repeat (STR) analysis (ACTG Inc.). RXDX106, Bemcentinib and Cabozantinib were purchased from Selleckchem, dissolved in DMSO at 10 mM concentration and diluted as necessary.

Majumder A., Hosseinian S., Stroud M., Adhikari E., Saller J.J., Smith M.A., Zhang G., Agarwal S., Creixell M., Meyer B.S., Kinose F., Bowers K., Fang B., Stewart P.A., Welsh E.A., Boyle T.A., Meyer A.S., Koomen J.M, & Haura E.B. (2022). Integrated proteomics-based physical and functional mapping of AXL kinase signaling pathways and inhibitors define its role in cell migration. Molecular cancer research : MCR, 20(4), 542-555.

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CCl4-Induced Liver Injury Model

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Wild-type C57BL/6 8- to 10-week-old male mice (Charles River, UK) were administered 0.4 mL/kg CCl₄ (Sigma-Aldrich), diluted in olive oil (Sigma-Aldrich) at a 1:3 ratio. Mice were injected intraperitoneally twice a week for total of 6 weeks (n = 13 intraperitoneal CCl₄ injections). In addition, a group of wild-type mice were administered bemcentinib (Selleckchem, cat. #S2841) intraperitoneally at 100 mg/kg dose 4 times over the last week before death. Twenty-four hours after the last CCl₄ injection, mice were killed, livers were perfused with PBS, and liver tissue was FFPE for fluorescent immunohistochemistry analysis. All animal experimental protocols were approved by Imperial College London in accordance with UK Home Office regulations (PPL numbers: 70/7578 and P8999BD42).

Pop O.T., Geng A., Flint E., Singanayagam A., Ercan C., Possamai L., Patel V.C., Kuenzler P., Meier M.A., Soysal S., Hruz P., Kollmar O., Tatham K.C., Ward J.K., Müllhaupt B., Weber A., Wendon J., Niess J.H., Heim M., Semela D., Weston C., Antoniades C.G., Terracciano L.M., Triantafyllou E., Brenig R.G, & Bernsmeier C. (2023). AXL Expression on Homeostatic Resident Liver Macrophages Is Reduced in Cirrhosis Following GAS6 Production by Hepatic Stellate Cells. Cellular and Molecular Gastroenterology and Hepatology, 16(1), 17-37.

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Pharmacological Inhibition of TGFBR1 and AXL

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The TGFBR1 inhibitor, galunisertib (LY2157299), and the AXL inhibitor, bemcentinib (R428), were purchased by Selleckchem. Drugs were dissolved in sterile dimethylsulfoxide (DMSO) at a 10 mM stock solution concentration and stored in aliquots at − 20 °C.

Ciardiello D., Blauensteiner B., Matrone N., Belli V., Mohr T., Vitiello P.P., Martini G., Poliero L., Cardone C., Napolitano S., De Falco V., Giunta E.F., Ciaramella V., Corte C.D., Barra G., Selvaggi F., Franco R., Marino F.Z., Cuomo A., Morgillo F., Troiani T., Sibilia M., Ciardiello F, & Martinelli E. (2021). Dual inhibition of TGFβ and AXL as a novel therapy for human colorectal adenocarcinoma with mesenchymal phenotype. Medical Oncology (Northwood, London, England), 38(3), 24.

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