Temsirolimus

Catalog No.S1044 Batch:S1044

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Technical Data

Formula

C56H87NO16
Molecular Weight 1030.29 CAS No. 162635-04-3
Solubility (25°C)* In vitro Ethanol 75 mg/mL (72.79 mM)
DMSO 67 mg/mL (65.03 mM)
Water Insoluble
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description Temsirolimus is a specific mTOR inhibitor with IC50 of 1.76 μM in a cell-free assay. Temsirolimus induces autophagy and apoptosis.
Targets
mTOR [1]
(Cell-free assay)
1.76 μM
In vitro In the absence of FKBP12, Temsirolimus potently inhibits mTOR kinase activity with IC50 of 1.76 μM, similar to that of rapamycin with IC50 of 1.74 μM. Temsirolimus treatment at nanomolar concentrations (10 nM to <5 μM) displays a modest and selective antiproliferative activity via FKBP12-dependent mechanism, but can completely inhibit the proliferation of a broad panel of tumor cells at low micromolar concentrations (5-15 μM), involving FKBP12-independent suppression of mTOR signaling. Temsirolimus treatment at micromolar but not nanomolar concentrations (20 μM) causes a marked decline in global protein synthesis and disassembly of polyribosomes, accompanied by rapid increase in the phosphorylation of translation elongation factor eEF2 and the translation initiation factor eIF2A. [1] Temsirolimus inhibits the phosphorylation of ribosomal protein S6, more potently in PTEN-positive DU145 cells than in PTEN-negative PC-3 cells, and inhibits cell growth and clonogenic survival of both cells in a concentration-dependent manner. [2] Temsirolimus (100 ng/mL) potently inhibits proliferation and induces apoptosis in primary human lymphoblastic leukemia (ALL) cells. [3]
In vivo In the NOD/SCID xenograft models with human ALL, Temsirolimus treatment at 10 mg/kg/day produces a decrease in peripheral blood blasts and in splenomegaly [3] Administration of Temsirolimus (20 mg/kg i.p. 5 days/week) significantly delays the growth of DAOY xenografts by 160% after 1 week and 240% after 2 weeks, compared with controls. Single high-dose of Temsirolimus (100 mg/kg i.p) treatment induces 37% regression of tumor volume within 1 week. Temsirolimus treatment for 2 weeks also delays the growth of rapamycin-resistant U251 xenografts by 148%. [4] Inhibition of mTOR by Temsirolimus improves performance on four different behavioral tasks and decreases aggregate formation in a mouse model of Huntington disease. [5] Administration of Temsirolimus induces significant dose-dependent, antitumor responses against subcutaneous growth of 8226, OPM-2, and U266 xenografts with ED50 of 20 mg/kg and 2 mg/kg for 8226 and OPM-2, respectively, which are associated with inhibited proliferation and angiogenesis, induction of apoptosis, and reduction in tumor cell size. [6]

Protocol (from reference)

Kinase Assay:

[1]
  • In vitro assay of mTOR catalytic activity

    The Flag-tagged wild-type human mTOR (Flag-mTOR) DNA constructs are transiently transfected into HEK293 cells. Protein extraction and purification of Flag-mTOR are carried out 48 hours later. In vitro kinase assays of purified Flag-mTOR in the presence of various concentrations of Temsirolimus without FKBP12 are performed in 96-well plate and detected by dissociation-enhanced lanthanide fluorescent immunoassay (DELFIA) using His6-S6K1 as the substrate. Enzymes is first diluted in kinase assay buffer (10 mM Hepes (pH 7.4), 50 mM NaCl, 50 mM β-glycerophosphate, 10 mM MnCl2, 0.5 mM DTT, 0.25 μM microcystin LR, and 100 μg/mL BSA). To each well, 12 μL of the diluted enzyme is mixed briefly with 0.5 μL Temsirolimus. The kinase reaction is initiated by adding 12.5 μL kinase assay buffer containing ATP and His6-S6K to give a final reaction volume of 25 μL containing 800 ng/mL FLAG-mTOR, 100 μM ATP, and 1.25 μM His6-S6K. The reaction plate is incubated for 2 hours (linear at 1-6 hours) at room temperature with gentle shaking and then terminated by adding 25 μL Stop buffer (20 mM Hepes (pH 7.4), 20 mM EDTA, and 20 mM EGTA). The DELFIA detection of the phosphorylated (Thr-389) His6-S6K is performed at room temperature using a monoclonal anti-P(T389)-p70S6K antibody labeled with Europium-N1-ITC (Eu) (10.4 Eu per antibody). 45 μL of the terminated kinase reaction mixture is transferred to a MaxiSorp plate containing 55 μL PBS. The His6-S6K is allowed to attach for 2 hours after which the wells are aspirated and washed once with PBS. 100 μL of DELFIA buffer with 40 ng/mL Eu-P(T389)-S6K antibody is added. The antibody binding is continued for 1 hour with gentle agitation. The wells are then aspirated and washed four times with PBS containing 0.05% Tween 20 (PBST). 100 μL of DELFIA Enhancement solution is added to each well and the plates are read in a PerkinElmer Victor model plate reader.
Cell Assay:

[1]
  • Cell lines

    A549, H157, H460, H446, HCT116, HT29, SW480, DLD1, Caco2, LNCap, DU145, MDA468, MDA231, HEK293, and PC3-MM2

  • Concentrations

    Dissolved in DMSO, final concentrations ~20 μM

  • Incubation Time

    72 hours

  • Method

    Cells are exposed to various concentrations of Temsirolimus for 72 hours. After treatment, viable cell densities are determined by MTS dye conversion using CellTiter AQ assay kit.
Animal Study:

[4]
  • Animal Models

    Female athymic nude mice injected s.c. with DAOY, or U251 cells

  • Dosages

    20 mg/kg

  • Administration

    Injection daily 5 times per week

Customer Product Validation

Data from [Data independently produced by Mol Cancer Res, 2014, 12, 703-13]

Data from [Data independently produced by PLoS One, 2013, 8, e62104]

Data from [Data independently produced by PLoS One, 2013, 8, e62104]

Data independently produced by , , Dr. Zhang of Tianjin Medical University

Selleck's Temsirolimus has been cited by 96 publications

Long-term statins administration exacerbates diabetic nephropathy via ectopic fat deposition in diabetic mice [ Nat Commun, 2023, 14(1):390] PubMed: 36693830
Unbiased evaluation of rapamycin's specificity as an mTOR inhibitor [ Aging Cell, 2023, e13888.] PubMed: 37222020
A cisplatin conjugate with tumor cell specificity exhibits antitumor effects in renal cancer models [ BMC Cancer, 2023, 23(1):499] PubMed: 37268911
Therapeutic Efficacy of Temsirolimus in a Patient-derived Model of Metastatic Fibrolamellar Hepatocellular Carcinoma [ In Vivo, 2023, 37(5):1940-1950] PubMed: 37652480
Proteo-genomic characterization of virus-associated liver cancers reveals potential subtypes and therapeutic targets [ Nat Commun, 2022, 13(1):6481] PubMed: 36309506
Rapalogs downmodulate intrinsic immunity and promote cell entry of SARS-CoV-2 [ J Clin Invest, 2022, 132(24)e160766] PubMed: 36264642
CIC-mediated modulation of MAPK signaling opposes receptor tyrosine kinase inhibitor response in kinase-addicted sarcoma [ Cancer Res, 2022, canres.1397.2021] PubMed: 35074756
JAK and mTOR inhibitors prevent cytokine release while retaining T cell bispecific antibody in vivo efficacy [ J Immunother Cancer, 2022, 10(1)e003766] PubMed: 35064010
The oncogene-dependent resistance to reprogramming unveils cancer therapeutic targets [ Cell Rep, 2022, 39(4):110721] PubMed: 35476996
GATOR2-dependent mTORC1 activity is a therapeutic vulnerability in FOXO1 fusion-positive rhabdomyosarcoma [ JCI Insight, 2022, 7(23)e162207] PubMed: 36282590

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SHIPPING AND STORAGE
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