Tosedostat (CHR2797)

Catalog No.S1522 Batch:S152203

Print

Technical Data

Formula

C21H30N2O6
Molecular Weight 406.47 CAS No. 238750-77-1
Solubility (25°C)* In vitro DMSO 81 mg/mL (199.27 mM)
Ethanol 50 mg/mL (123.01 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.)

Preparing Stock Solutions

Biological Activity

Description Tosedostat (CHR2797) is an aminopeptidase inhibitor for LAP, PuSA and Aminopeptidase N with IC50 of 100 nM, 150 nM and 220 nM, respectively, and does not effectively inhibit either PILSAP, MetAP-2, LTA4 hydrolase, or MetAP-2. Phase 2.
Targets
LAP [1] PuSA [1] Aminopeptidase N [1]
100 nM 150 nM 220 nM
In vitro CHR-2797 has almost no effect on Aminopeptidase B, PILSAP, LTA4 hydrolase and MetAP-2 activity with IC50 values of >1 uM, >5 uM, >10 uM and >30 uM, respectively. CHR-2797 is converted into a pharmacologically active acid product (CHR-79888) inside cells, which shows significant inhibitory activity towards LTA4 hydrolase with IC50 of 8 nM. CHR-2797 exhibits profound anti-proliferative effects against a range of cancer cell lines such as U-937, HL-60, KG-1 and GDM-1 with IC50 values of 10 nM, 30 nM, 15 nM and 15 nM, respectively, but is inactive against HuT 78 and Jurkat E6-1 with IC50 values of >10 uM. There is no obvious correlation between sensitivity to CHR-2797 and the mutational status of p53, PTEN, or K-Ras in cells. CHR-2797 shows selectivity for transformed cells (MrC5-SV2 or K-ras NRK) over non-transformed cells (MrC5 or NRK). CHR-2797 (6 μM) treatment leads to the up-regulation of genes involved in amino acid transport and metabolic pathways, the phosphorylation of eukaryotic initiation factor 2α, the inhibition of phosphorylation of mTOR substrates and reduced protein synthesis in HL-60 cells. [1]
In vivo Administration of CHR-2797 (~100 mg/kg) decreases tumor volumes in vivo, compared to controls, in a dose-response manner in the rat HOSP.1 lung colonisation model, the rat HSN LV10 chondrosarcoma liver colonisation model, the human MDA-MB-435 breast cancer spontaneous metastasis model, and the human MDA-MB-468 cell xenograft model. [1]

Protocol (from reference)

Kinase Assay:[1]
  • Aminopeptidase assays

    LAP activity is determined by measuring the hydrolysis of the tripeptide, LGG, which is detected by the derivatization reagent OPA in the presence of β-mercaptoethanol. The assay is carried out in 96-well assay plates. Wells contain diluted CHR-2797 (5 μL), 10 μg/mL LAP (5 μL) and 40 μL of 0.5 mM LGG. The plate is shaken briefly, and incubated for 90 minutes at 37 °C. The reaction is terminated by addition of 200 μL of OPA/β-Mercaptoethanol per well. The plate is read on the Victor Wallac3 plate reader: excitation, 355nm and emission, 460nm. PuSA activity is determined using the fluorogenic substrate Ala-AMC. Incubation mix contains diluted CHR-2797 (20 μL), substrate (125 μM Ala-AMC in 0.125 M Tris-HCl buffer, pH 7.5; 40 μL) and enzyme (40 μL). After incubation for 2 hours at 37 °C, the reaction is stopped by addition of 100 μL 3% (v/v) acetic acid. Fluorescence is measured using a SLT Fluostar fluorimeter. Aminopeptidase N is assayed using the fluorogenic substrate, Ala-AMC. Incubation mix contains diluted CHR-2797 (20 μL), substrate (40 μL; final concentration, 60 μM) and enzyme (40 μL, 1:8000 dilution) and is incubated for 60 minutes at 37 °C prior to addition of 100 μL 3% (v/v) acetic acid, to stop the reaction. Fluorescence is measured using a SLT Fluostar fluorimeter.
Cell Assay:[1]
  • Cell lines

    U-937, HL-60, KG-1, HNT-34 and GDM-1

  • Concentrations

    Dissolved in DMSO, final concentration ~1 mM

  • Incubation Time

    72 hours

  • Method

    Cells are exposed to different concentrations of CHR-2797 for 72 hours. During the final 4 hours of this incubation, cells are pulsed with 0.4 μCi/well of [3H]thymidine (specific activity, 5 mCi/mmol), harvested onto GF/C glass fiber filter mats using a Tomtec harvester, and counted on a 1450 MicroBeta scintillation counter to determine the amount of [3H]thymidine incorporated into cellular DNA. Inhibition of the proliferation of human cancer cell lines is measured by [3H]thymidine incorporation.
Animal Study:[1]
  • Animal Models

    Female rats (CBH/cbi) injected i.v. with HOSP.1P cells, male (CBH/cbi) rats injected with HSN LV10 cells, and nude mice (MF1 (nu/nu) inoculated with MDA-MB 435 cells or MDA-MB-468 cells

  • Dosages

    ~100 mg/kg

  • Administration

    Dosed daily p.o.

Selleck's Tosedostat (CHR2797) has been cited by 3 publications

Predicting effective pro-apoptotic anti-leukaemic drug combinations using co-operative dynamic BH3 profiling [ PLoS One, 2018, 13(1):e0190682] PubMed: 29298347
Response of Single Leukemic Cells to Peptidase Inhibitor Therapy Across Time and Dose Using a Microfluidic Device [Kovarik ML ,et al. Integr Biol, 2014, 6(2):164-74] PubMed: 24413844
Microfluidic chemical cytometry of peptide degradation in single drug-treated acute myeloid leukemia cells. [Kovarik ML, et al. Anal Chem, 2013, 85(10):4991-7] PubMed: 23590517

RETURN POLICY
Selleck Chemical’s Unconditional Return Policy ensures a smooth online shopping experience for our customers. If you are in any way unsatisfied with your purchase, you may return any item(s) within 7 days of receiving it. In the event of product quality issues, either protocol related or product related problems, you may return any item(s) within 365 days from the original purchase date. Please follow the instructions below when returning products.

SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.