In vitro comparison of O4-benzylfolate modulated, BCNU-induced toxicity in human bone marrow using CFU-GM and tumor cell lines.
2-Amino-O4-benzylpteridine derivatives inactivate the human DNA repair protein O6-alkylguanine-DNA alkyltransferase and have been tested as modulators of alkylating agent chemotherapy. Recently, the therapeutic potential of O4-benzylfolate (O4BF) in modulating bis-chloroethylnitrosourea (BCNU) toxicity was demonstrated in vitro using human HT29 and KB tumor lines. The current studies replicated the previous findings in HT29 and KB cells using ATP as an endpoint. However, the effective treatment conditions were severely toxic to human neutrophil progenitors called CFU-granulocyte/macrophage (CFU-GM), which could not tolerate > or =40 microM BCNU at any O4BF concentration. A lower BCNU concentration (10 microM) in combination with O4BF (2-100 microM) was only moderately tumoricidal. To screen for conditions tolerated by CFU-GM, bone marrow (BM) cells were pre-incubated (5 h) with O4BF, co-treated with O4BF and BCNU (42 h), washed, and plated to quantify CFU-GM survival. O4BF at 2 or 5 microM progressively lowered the inhibitory concentrations (ICs) for BCNU, but further increases in O4BF concentrations did not. Increasing O4BF concentrations with constant BCNU (10 microM) under the same prolonged exposure as in the human marrow study achieved only modest tumoricidal effects. In summary, the unexpected finding that normal BM cells are impacted by an agent developed to target malignant tissue refutes speculation that normal beta-folate receptor expressing hematopoietic cells will be spared. Further, the validated IC90 endpoint from the huCFU-GM assay has provided a reference point for judging the potential therapeutic effectiveness of this investigational combination in man using in vitro assays.
Behrsing
Holger Peter
HP
Predictive Toxicology Section, Laboratory for Human Toxicology and Pharmacology, Applied and Developmental Research Directorate, SAIC-Frederick, Inc., NCI-Frederick, Building 438, Ft. Detrick, Frederick, MD 21702, USA. behrsingh@mail.nih.gov
Furniss
Michael J
MJ
Robillard
Kristine A
KA
Tomaszewski
Joseph E
JE
Parchment
Ralph E
RE
D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000187:drug effects*; D002330:Carmustine / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005492:Folic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D055014:Granulocyte-Macrophage Progenitor Cells / Q000187:drug effects*; D019073:HT29 Cells; D006801:Humans; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000037:antagonists & inhibitors; D013997:Time Factors
D018906:Antineoplastic Agents, Alkylating; C524118:O(4)-benzylfolic acid; D005492:Folic Acid; D019853:O(6)-Methylguanine-DNA Methyltransferase; D002330:Carmustine