Ganetespib

Manufactured by Selleck Chemicals

Sourced in United States

Ganetespib is a heat shock protein 90 (Hsp90) inhibitor. It is a synthetic small molecule that binds to and inhibits the activity of Hsp90, a chaperone protein essential for the stability and function of various client proteins involved in cell signaling and proliferation.

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Lab products found in correlation

Luminespib, by Selleck Chemicals (4 mentions) Cycloheximide (chx), by Merck Group (4 mentions) Palbociclib, by Selleck Chemicals (3 mentions) Bortezomib, by Selleck Chemicals (3 mentions) Auy922, by Selleck Chemicals (2 mentions) Nms e973, by Selleck Chemicals (2 mentions) Ribociclib, by MedChemExpress (2 mentions) Doxycycline, by Merck Group (2 mentions) Blasticidin s, by Merck Group (2 mentions) Abemaciclib, by MedChemExpress (2 mentions) Mln4924, by MedChemExpress (2 mentions) Alectinib, by Selleck Chemicals (2 mentions) Crizotinib, by Selleck Chemicals (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Mda mb 453, by Thermo Fisher Scientific (1 mentions) Mda mb 468, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions) Foetal bovine serum (fbs), by Merck Group (1 mentions)

Spelling variants of this product

Ganetespib (STA-9090) (2 citations)

22 protocols using ganetespib

Breast Cancer Cell Line Characterization

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MDA-MB-231, MDA-MB-468 and MDA-MB-453 cell lines were obtained from the American Type Culture Collection. HCC-1143, HCC-1937 and Hs578T cell lines were obtained from Dr. P McGowan (St Vincent’s University Hospital, Ireland) while BT-20 cell line was obtained from Professor Adrian Harris (University of Oxford, UK). MDA-MB-453, MDA-MB-468, MDA-MB-231, Hs578T and BT-20 cell lines were maintained in DMEM (Dulbecco’s modified Eagle medium; Life Technologies), supplemented with 10% (v/v) foetal bovine serum (FBS; Sigma-Aldrich), 2 mM GlutaMax (Life Technologies) and 50 U penicillin/ 50 μg/ml streptomycin (Life Technologies). HCC-1143 and HCC-1937 cell lines were maintained in RPMI-1640 medium (Life Technologies), supplemented with 10% (v/v) FBS, 2 mM GlutaMax and 50 U penicillin/ 50 μg/ml streptomycin. The cell lines were authenticated using Short Tandem Repeat (STR) analysis and tested for Mycoplasma infection using PCR, as described in [32 (link)].
Ganetespib and a 326-compound bioactive small molecule library (L1100), including 17-AAG and NVP-AUY922, were obtained from Selleckchem (USA). All stock solutions were prepared DMSO and stored at − 20 °C.

Mumin N.H., Drobnitzky N., Patel A., Lourenco L.M., Cahill F.F., Jiang Y., Kong A, & Ryan A.J. (2019). Overcoming acquired resistance to HSP90 inhibition by targeting JAK-STAT signalling in triple-negative breast cancer. BMC Cancer, 19, 102.

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Cell Line Characterization and Drug Treatments

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Ramos, Raji, Namalwa, Daudi, CA-46, Ly10 and Ly18 cell lines were purchased from American Type Culture Collection (ATCC) in 2013. Jiyoye and DG-75 were purchased from ATCC in 2014. Raji 2R and Raji 4RH were provided by Roswell Park Cancer Institute (M.J.B.). All cells were used for experiments within 2 months of thawing. Cell line characterization was performed by Idex Bioresearch (Westbrook, Maine) using short tandem repeat profiling and multiplex PCR to report cell line characterization and interspecies contamination. Testing was last performed in February 2014. Cells were cultured in RPMI-1640 media (Invitrogen) supplemented with 10% heat inactivated fetal bovine serum and gentamicin 50ug/mL (Sigma-Aldrich). PU-H71, PU-H71-beads, and control beads were synthesized and prepared at Memorial Sloan Kettering Cancer Center as previously reported(13 (link),14 (link)). Idelalisib (CAL-101), IPI-145, BKM120 (15 (link)), BEZ-235 (16 (link)), ganetespib (17 (link)), and BIIB-021 (18 (link)) were purchased from Selleck Chemicals. MPC-3100 (19 (link)) and CUDC-305 (20 (link)) were purchased from Santa Cruz and ChemieTek respectively. The chemical structures for PU-H71 and BEZ-235 are presented in Supplemental Figure 1.

Giulino-Roth L., van Besien H.J., Dalton T., Totonchy J.E., Rodina A., Taldone T., Bolaender A., Erdjument-Bromage H., Sadek J., Chadburn A., Barth M.J., Dela Cruz F.S., Rainey A., Kung A.L., Chiosis G, & Cesarman E. (2017). Inhibition of Hsp90 suppresses PI3K/AKT/mTOR signaling and has antitumor activity in Burkitt lymphoma. Molecular cancer therapeutics, 16(9), 1779-1790.

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Preclinical Evaluation of HER2-Targeted Therapies

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For preclinical studies, T-DM1 was obtained from MSK pharmacy and T-DXd was obtained from Daichii Sankyo, Co., Ltd. The HER2 inhibitors neratinib (provided by Puma Biotechnology), afatinib (Selleckchem, #S1011), lapatinib (Selleckchem, # S1028) and tucatinib (Selleckchem, #S8362) and the HSP90 inhibitor ganetespib (Selleckchem, #S1159) were dissolved in DMSO (10 mM stock), aliquoted and stored at −20℃. The endocytosis inhibitor dynasore (Abcam, #ab120192) was dissolved in DMSO (30 mM stock), aliquoted and stored at −20°C.

Li B.T., Michelini F., Misale S., Cocco E., Baldino L., Cai Y., Shifman S., Tu H.Y., Myers M.L., Xu C., Mattar M., Khodos I., Little M., Qeriqi B., Weitsman G., Wilhem C.J., Lalani A.S., Diala I., Freedman R.A., Lin N.U., Solit D.B., Berger M.F., Barber P.R., Ng T., Offin M., Isbell J.M., Jones D.R., Yu H.A., Thyparambil S., Liao W.L., Bhalkikar A., Cecchi F., Hyman D.M., Lewis J.S., Buonocore D.J., Ho A.L., Makker V., Reis-Filho J.S., Razavi P., Arcila M.E., Kris M.G., Poirier J.T., Shen R., Tsurutani J., Ulaner G.A., de Stanchina E., Rosen N., Rudin C.M, & Scaltriti M. (2020). HER2-mediated internalization of cytotoxic agents in ERBB2 amplified or mutant lung cancers. Cancer discovery, 10(5), 674-687.

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Cell Line Characterization and Compound Screening

Cited 1 time

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HepG2 (SCSP-510), Huh7 (SCSP-526), and L02 (GNHu 6) were purchased from the Cell Bank, Chinese Academy of Medical Sciences (Shanghai, China). MHCC97H were obtained from the Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, China. HepG2-Luc were constructed by Cloud-Clone Corp (Wuhan, China). All cell lines were authenticated, and cells were thawed upon arrival, expanded, and stored in a liquid nitrogen tank. The absence of mycoplasma cell contamination was confirmed using MycAwayTM-Color One-Step Mycoplasma Detection Kit (40611ES, Yeasen, China). All the cell lines were cultured in Dulbecco’s modified Eagle’s medium (Gibco, USA) supplemented with 10% fetal bovine serum (Gibco, USA) at 37°C, 5% CO₂. Ganetespib (STA9090) (22 (link)) (S1159), Novobiocin (NB) (23 (link)) (S2492), NVP-BEP800 (24 (link)) (S1498), MG132 (S2619), and cycloheximide (CHX) (S7418) were purchased from SelleckChem (USA).

Liu L., Deng Y., Zheng Z., Deng Z., Zhang J., Li J., Liang M., Zhou X., Tan W., Yang H., Neckers L.M., Zou F, & Chen X. (2021). Hsp90 Inhibitor STA9090 Sensitizes Hepatocellular Carcinoma to Hyperthermia-Induced DNA Damage by Suppressing DNA-PKcs Protein Stability and mRNA Transcription. Molecular cancer therapeutics, 20(10), 1880-1892.

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Hsp90 Inhibitors and Cell Lines

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The Hsp90 inhibitors SNX-2112 (S2639, >99%) AUY922 (S1069, 99%) ganetespib (S1159, >99%) were purchased from Selleck Chemicals. Geldanamycin (ant-gl-5, >95%) was purchased from Invivogen. Fluconazole was obtained from Sequoia Research Products (SRP01025f, >99%) and Cy3B-Mono NHS ester was purchased from GE Healthcare UK Ltd (PA63101, >95%). The cell lines HEPG2 (HB-8065), Raw264.5 (TIB-71), 293T (CRL-3216), and 3T3 (NIH/3T3; CRL-1658) were obtained from the American Type Culture Collection (ATCC) and grown in DMEM supplemented with 10% fetal bovine serum at 37 °C under 5% CO₂. Cultures were confirmed negative for mycoplasma contamination by monthly surveillance testing using a PCR-based kit.

Whitesell L., Robbins N., Huang D.S., McLellan C.A., Shekhar-Guturja T., LeBlanc E.V., Nation C.S., Hui R., Hutchinson A., Collins C., Chatterjee S., Trilles R., Xie J.L., Krysan D.J., Lindquist S., Porco JA J.r., Tatu U., Brown L.E., Pizarro J, & Cowen L.E. (2019). Structural basis for species-selective targeting of Hsp90 in a pathogenic fungus. Nature Communications, 10, 402.

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HeLa Cell Maintenance and Transfection

Cited 2 times

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HeLa cells were maintained in TumorPlus 263 medium (Capricorn Scientific GmbH, Ebersdorfergrund, Germany) or RPMI1640 supplemented 10% fetal calf serum at 37 °C and 5% CO₂. Cells were transfected with plasmid DNA using Fugene HD (Promega, Mannheim, Germany). HeLa cells with stable expression of CDC37-nanoKAZ from the pCAGIPuro vector^{15 (link)} were selected with puromycin for 2 weeks. If indicated, cells were treated with either ganetespib (Selleck Chemicals, Houston, TX, USA) dissolved in DMSO. Rat RINm5F insulinoma cells were available in the lab from earlier projects and grown in RPMI 1640 medium as described^{41 (link)}.

Abu Jhaisha S., Widowati E.W., Kii I., Sonamoto R., Knapp S., Papadopoulos C, & Becker W. (2017). DYRK1B mutations associated with metabolic syndrome impair the chaperone-dependent maturation of the kinase domain. Scientific Reports, 7, 6420.

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Investigating HSP90 Inhibitor-Mediated Glycolytic Changes

Cited 1 time

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HSP90 inhibitors 17-AAG, luminespib, and ganetespib were obtained from SelleckChem (Houston, TX). CHX and MG132 were purchased from Sigma-Aldrich (St. Louis, MO), and rhIL-8 was purchased from PeproTech (Rocky Hill, NJ). MTS Assay Kit was obtained from Abcam (Cambridge, UK). Glycolysis antibody sampler kit, including PKM2, PFKP, glyceraldehyde-3-phosphate dehydrogenase, hexokinase I (HKI), HKII, lactate dehydrogenase A (LDHA), and PDH antibodies, was purchased from Cell Signaling Technology (Beverly, MA). Antibodies that recognize Ub, PDL1, and IL-8 were purchased from Novus Biologicals (Littleton, CO). Antibodies against IL-1α and IL-1β were ordered from Abcam, and antibodies against HSP90 and β-actin were obtained from Sigma-Aldrich (St. Louis, MO). Antibody that recognizes CD8α was purchased from Cell Signaling Technology. Secondary antibodies were purchased from Invitrogen (Carlsbad, CA). Cell proliferation, viability, colony formation, and Western blot assays were carried out as previously described (18 (link), 43 (link)).

Chen F., Tang C., Yang F., Ekpenyong A., Qin R., Xie J., Momen-Heravi F., Saba N.F, & Teng Y. (2024). HSP90 inhibition suppresses tumor glycolytic flux to potentiate the therapeutic efficacy of radiotherapy for head and neck cancer. Science Advances, 10(8), eadk3663.

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Synthesis of 5-Isoxazole Carboxamide Inhibitor

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Synthesis of the precursor compound 7, 5-[2,4-Dihydroxy-6-(4-nitrophenoxy) phenyl]-N-(piperidin-4-yl)-isoxazole-3-carboxamide was performed as described by Brasca et al.26 and a detailed synthetic protocol and characterization is provided in Supplementary Material (Figure S1). HSP90 inhibitors, Ganetespib, PU-H71, Pifithrin-µ, PU-WS13, and authentic reference compound NMS-E973 were purchased from Selleckchem or MedChem Express and used without further purification. Total polar surface area (tPSA) and LogD values were calculated using Marvinsketch (Marvin 14.10.13.0, 2014, ChemAxon, http://www.chemaxon.com).

Vermeulen K., Naus E., Ahamed M., Attili B., Siemons M., Luyten K., Celen S., Schymkowitz J., Rousseau F, & Bormans G. (2019). Evaluation of [11C]NMS-E973 as a PET tracer for in vivo visualisation of HSP90. Theranostics, 9(2), 554-572.

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Acquisition and Preparation of Compounds

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Commercially available compounds Palbociclib, Ganetespib, Luminespib and Bortezomib were obtained from Selleckchem. Abemaciclib, Ribociclib and MLN4924 were obtained from Medchem Express. Blasticidin S, cycloheximide and doxycycline were purchased from Sigma. Chemical compounds were dissolved either in DMSO or water and stored at −20 °C.

Wu X., Yang X., Xiong Y., Li R., Ito T., Ahmed T.A., Karoulia Z., Adamopoulos C., Wang H., Wang L., Xie L., Liu J., Ueberheide B., Aaronson S.A., Chen X., Buchanan S.G., Sellers W.R., Jin J, & Poulikakos P.I. (2021). Distinct CDK6 complexes determine tumor cell response to CDK4/6 inhibitors and degraders. Nature cancer, 2(4), 429-443.

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Generation and Characterization of PU-H71-Resistant Cell Lines

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All cell lines were obtained from ATCC and maintained under standard conditions. Cell line identity and purity were verified using the Multiplex Cell Authentication and Contamination Tests (Multiplexion). PU-H71 was generated and provided by the laboratory of G. Chiosis as previously described [47 (link)]. Puromycin was obtained from Sigma-Aldrich. Ganetespib, tanespimycin and tariquidar were purchased from Selleck. From each PU-H71-resistant cell line, three to five clones were generated by seeding single cells into 96-well plates.

Rouhi A., Miller C., Grasedieck S., Reinhart S., Stolze B., Döhner H., Kuchenbauer F., Bullinger L., Fröhling S, & Scholl C. (2016). Prospective identification of resistance mechanisms to HSP90 inhibition in KRAS mutant cancer cells. Oncotarget, 8(5), 7678-7690.

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