(±)Amethopterin hydrate 

Add a minimal amount of 0.1 N NaOH to solubilize, then dilute in neutral buffer or saline. Diluted stock is stable for 1 week refrigerated and for 1 month frozen at −20 °C. References: Cell & Tissue Culture: Laboratory Procedures, Chapter 27, Page 27D:3.2 (A. Doyle, et al., eds, John Wiley) and Cell Biology: A Laboratory Handbook, vol. 2, page 417 (Julio E. Celis, ed., Academic Press).

Folic acid antagonist and potent anticancer agent. Blocks DNA synthesis by blocking the production of tetrahydrofolate covactors required for the synthesis of thymidine. Amethopterin is actively transported into cells by the folate transporter. In the cell, it is converted to a high molecular weight polyglutamate metabolite by folylpolyglutamate synthase that, in turn, binds to dihydrofolate reductase and inhibits its activity. Amethopterin-polyglutamate is degraded intracellularly by γ-glutamyl hydrolase.

≥95%

powder

yellow to yellow with an orange cast

H₂O: insoluble

NaOH: minimal amount of 0.1 N NaOH to solubilize, then dilute in neutral buffer or salinesoluble

human ... DHFRP1(573971), FPGS(2356), SLC19A1(6573), TYMS(7298)

rat ... Dhfr(24312)

mouse ... Dhfr(13361)

−20°C

Sharif, K.A., et al., Concentrating capacity of the human reduced folate carrier (hRFC1) in human ZR-75 breast cancer cell lines. Biochem. Pharmacol. 55, 1683-1689, (1998) abstract

Doyle, J.B., et al. Cell and Tissue Culture: Laboratory Procedures, 27D

Yao, R., et al., Human γ-glutamyl hydrolase: cloning and characterization of the enzyme expressed in vitro. Proc. Natl. Acad. Sci. USA 93, 10134-10138, (1996) abstract

Jansen, G., et al., Regulation of carrier-mediated transport of folates and antifolates in methotrexate-sensitive and -resistant leukemia cells. Adv. Enzyme Regul. 37, 59-76, (1997) abstract

Kumar, P., et al., Interaction of polyglutamyl derivatives of methotrexate, 10-deazaaminopterin, and dihydrofolate with dihydrofolate reductase. Cancer Res. 46, 5020-5023, (1986) abstract

Kumar, P., et al., Inhibition of human dihydrofolate reductase by antifolyl polyglutamates. Biochem. Pharmacol. 38, 541-543, (1989) abstract

Rhee, M.S., et al., Characterization of human cellular γ-glutamyl hydrolase. Mol. Pharmacol. 53, 1040-1046, (1998) abstract

Longo, G. S., et al., γ-Glutamyl hydrolase and folylpolyglutamate synthetase activities predict polyglutamylation of methotrexate in acute leukemias. Oncol. Res. 9, 259-263, (1997) abstract

Aldrich MSDS 1, 54:D / Corp MSDS 1 (1), 129:B / FT-IR 2 (3), 3973:A / FT-IR 1 (2), 898:B / IR-Spectra (3), 1416:F / RegBook 1 (3), 3539:D / RegBook 1 (2), 2663:G / Sigma FT-IR 1 (2), 252:C / Structure Index 1, 419:B:1

T

61-25-36/38

53-26-36/37-45

UN 1544 6.1/PG 2

3

MA1224100