In February 1948, Sidney Farber, a pathologist at Harvard Medical School, began experiments with the antifolate drug aminopterin
. This early chemotherapy drug, and its successor methotrexate, had been synthesized by Yellapragada Subbarrow, an Indian chemist who led the research program at Lederle Labs, along with his colleague Harriet Kiltie.
NG108-15 cells (obtained from China Center for Type Culture Collection (CCTCC), Wuhan, China) were cultured in Dulbecco's modified essential media (DMEM) (GIBCO) supplemented with 10% (v/v) FBS (GIBCO) and 1 x HAT (100 [micro]M hypoxanthine, 0.4 [micro]M aminopterin
, and 16 [micro]M thymidine) (H0262, Sigma).
With the identification of the beneficial effects of aminopterin
antimetabolite, chemotherapeutic agents known as disease-modifying anti-rheumatic drugs (DMARDs) were introduced to RA therapeutics in late 1980s.
This procedure was repeated twice and the aminopterin
concentration thus continuously reduced over six days, while hypoxanthine and thymidine were supplemented following manufacturer's recommendations (1: 100).
Wolff, "Temporary remissions in acute leukemia in children produced by folic acid antagonist, 4-aminopteroyl-glutamic acid (aminopterin
)," New England Journal of Medicine, vol.
To remove background mutants, MCL-5 cells were cultured for 3 days in R10 containing HAT [hypoxanthine, aminopterin
, thymidine; Hybri-Max[TM] (Sigma-Aldrich, Poole, UK)], which is lethal to cells harboring mutations at the TK locus (Busby et al.
* + + / [+ or -] + Azathioprine 0 0 5-fluorouracil * + + + / + + + Methotrexate * + + / [+ or -] 0 Thioguanine + + No activity shown by cytarabine, mercaptopurine.
Sidney Farber, a pathologist at Boston Children's Hospital, was so distressed doing autopsies on these children that he moved into the clinic and, against the advice of more conservative colleagues, began treating children with aminopterin
, a highly toxic drug that starved their cancerous white blood cells of critical nutrients.
The isosteric univalent replacement of hydroxal group by amino group can be illustrated with 4-amino-4-deoxyderivative (aminopterin
), its N10- and its N10-Methyl derivative (methotrexate) and folic acid.
Hybrid cells were selected in growth medium supplemented with hypoxanthine, aminopterin
, and thymidine (50x HAT media supplement, Sigma-Aldrich Co.).
Examples include low folate intake levels and risk of NTDs was high in pregnancies from low socio-economic families [41, 42], mean RBC folate concentrations in women with a NTD was lower , folic acid metabolism in pregnant women affected by NTD was impaired , and the use of aminopterin
, a powerful folic acid antagonist, was associated with anencephaly .