Pemetrexed_disodium_DataSheet_MedChemExpress

Inhibitors, Agonists, Screening Libraries

https://www.wendangku.net/doc/dc2955443.html, Data Sheet

BIOLOGICAL ACTIVITY:

Pemetrexed disodium is a novel antifolate , the K i values of the pentaglutamate of LY231514 are 1.3, 7.2, and 65 nM for inhibits thymidylate synthase (TS ), dihydrofolate reductase (DHFR ), and glycinamide ribonucleotide formyltransferase (GARFT ), respectively.IC50 & Target: Ki: 1.3 nM (TS), 7.2 nM (DHFR), 65 nM (GARFT)[1]

In Vitro: Pemetrexed (LY231514) disodium is a novel classical antifolate, the antitumor activity of which may result from

simultaneous and multipie inhibition of several key folate–requiring enzymes via its polyglutamated metabolites. Pemetrexed (LY231514) is one of the best substrates that is known for the enzyme FPGS (K m =1.6 μM and V max /K m =621). It is likely that

polyglutamation and the polyglutamated metabolites of LY231514 play profound roles in determining both the selectivity and the antitumor activity of this novel agent. Whereas LY23l5l4 only moderately inhibits TS (K i =340 nM, recombinant mouse), the pentaglutamate of LY23l5l4 is 100–fold more potent (K i =3.4 nM), making LY231514 one of the most potent folate–based TS inhibitors [1].

In Vivo: The group of mice treated with PC61 plus Pemetrexed demonstrates statistically longer survival than other groups. In a survival analysis, significantly better survival is observed in the group of mice treated with PC61 plus Pemetrexed compare with those treated with PC61 alone, rat IgG plus Pemetrexed, or no treatment [2].

PROTOCOL (Extracted from published papers and Only for reference)

Kinase Assay:[1]AICARFT inhibition assays are carried out at room temperature by monitoring the formation of

[6S]–5,6,7,8–tetrahydrofolate from 10–formyl–[6R,S]–5,6,7,8–tetrahydrofolate at A 298. All solutions are purged with N 2 gas prior to use. The reaction solution contains 33 mM Tris–Cl, pH 7.4, 25 mM KCl, 5 mM 2–Mercaptoethanol, 0.05 mM AICA ribonucleotide, and 16 nM (2 milliunits/mL) of AICARFT. 10–Formyl–[6R,S]–5,6,7,8–tetrahydrofolate concentrations of 0.037, 0.074, and 0.145 mM are used (0.61, 1.23, and 2.45 times its K m value, respectively). LY231514 is tested as an inhibitor at 0.08–0.8 mM (four concentrations).When the tri– and pentaglutamates of LY231514 are used as inhibitors, the concentrations are 0.0005–0.009 mM (eight

concentrations). Enzyme assays are initiated by the addition of enzyme. Data is analyzed using the ENZFITTER program for competitive inhibition.

Cell Assay: Pemetrexed is dissolved in DMSO and stored, and then diluted with cell culture medium before use [1].[1]Dose–response curves are generated to determine the concentration required for 50% inhibition of growth (IC 50). Pemetrexed is dissolved initially in DMSO at a concentration of 4 mg/mL and further diluted with cell culture medium to the desired concentration. CCRF–CEM leukemia cells in complete medium are added to 24–well Cluster plates at a final concentration of 4.8×l04 cells/well in a total volume of 2 mL.Test compounds at various concentrations are added to duplicate wells so that the final volume of DMSO is 0.5%. The plates are incubated for 72 h at 37°C in an atmosphere of 5% CO 2 in air. At the end of the incubation, cell numbers are determined on a ZBI Coulter counter. Control wells usually contain 4×105 to 6×105 cells at the end of the incubation. For several studies, IC 50s are

Product Name:

Pemetrexed (disodium)Cat. No.:

HY-10820A CAS No.:

150399-23-8Molecular Formula:

C 20H 19N 5Na 2O 6Molecular Weight:

471.37Target:

Antifolate; Autophagy Pathway:

Cell Cycle/DNA Damage; Autophagy Solubility:H 2

O: 11.19 mg/mL (Need ultrasonic and warming)

determined for each compound in the presence of either 300 μM AICA, 5 μM thymidine, 100 μM hypoxanthine, or combination of 5μM hymidine plus 100 μM hypoxanthine[1].

Animal Administration: Pemetrexed disodium is prepared in saline[2].[2]Mice[2]

Female CBA mice and female NOD/SCID mice (NOD.CB17–Prkdc scid) at 6–8 wk of age are used. Premetrexed (100 mg/kg) is given i.p. from days 4–8 (5 consecutive d) to tumor–bearing mice to explore the synergistic effect when combined with anti–CD25 Ab or IgG control. The dose and schedule used for Pemetrexed in the current study is determined based on previous studies in mice.

References:

[1]. Shih C, et al. LY231514, a pyrrolo[2,3–d]pyrimidine–based antifolate that inhibits multiple folate–requiring enzymes. Cancer Res. 1997 Mar 15;

57(6):1116–23.

[2]. Anraku M, et al. Synergistic antitumor effects of regulatory T cell blockade combined with pemetrexed in murine malignantmesothelioma. J Immunol. 2010 Jul 15;185(2):956–66.

Caution: Product has not been fully validated for medical applications. For research use only.

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